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DESCRIPCION

DESCRIPTION

Metodos y composiciones relacionados con enzimas tioesterasas

Methods and compositions related to thioesterase enzymes

Campo de la invencion

Field of the Invention

La presente divulgacion se refiere a una nueva enzima tioesterasa, a celulas hospedadoras recombinantes que comprenden la nueva tioesterasa, a metodos de produccion de derivados de acidos grasos y a derivados de acidos grasos producidos de ese modo y a usos de los mismos. Un aspecto particular de la presente divulgacion se refiere a la produccion de productos quimicos industriales y combustibles.

The present disclosure relates to a new thioesterase enzyme, to recombinant host cells comprising the new thioesterase, to methods of producing fatty acid derivatives and to fatty acid derivatives produced in this way and to uses thereof. A particular aspect of the present disclosure refers to the production of industrial chemicals and fuels.

Antecedentes de la invencion

Background of the invention

Los avances en tecnologia han venido acompanados por una mayor dependencia de combustibles y productos quimicos industriales de fuentes petroquimicas. Dichas fuentes de combustibles son cada vez mas limitadas y dificiles de adquirir. Con la quema de combustibles fosiles que tiene lugar a un ritmo sin precedentes, es probable que la demanda mundial de combustibles y de productos quimicos procedentes de fuentes petroquimicas supere pronto los suministros actuales.

Advances in technology have been accompanied by a greater dependence on fuels and industrial chemicals from petrochemical sources. These fuel sources are increasingly limited and difficult to acquire. With the burning of fossil fuels that takes place at an unprecedented rate, global demand for fuels and chemicals from petrochemical sources is likely to soon exceed current supplies.

Como resultado, se han dirigido esfuerzos hacia el aprovechamiento de las fuentes de energia renovable, tales como la luz del sol, el agua, el viento y la biomasa. El uso de biomasa para producir nuevas fuentes de combustible y productos quimicos no procedentes de fuentes de petroleo (por ejemplo, biocombustible), ha surgido como una opcion alternativa.

As a result, efforts have been directed towards the use of renewable energy sources, such as sunlight, water, wind and biomass. The use of biomass to produce new sources of fuel and chemicals not from oil sources (eg biofuel), has emerged as an alternative option.

El biocombustible es un combustible biodegradable y de combustion limpia que puede estar compuesto por alcanos y/o esteres. Un biocombustible ejemplar es el biodiesel. El biodiesel puede utilizarse en la mayoria de los motores diesel de combustion interna en forma pura, que se conoce como biodiesel "no diluido", o como una mezcla en cualquier concentracion con diesel de petroleo regular u otros biodiesel.

Biofuel is a biodegradable and clean combustion fuel that can be composed of alkanes and / or esters. An exemplary biofuel is biodiesel. Biodiesel can be used in most internal combustion diesel engines in pure form, known as "undiluted" biodiesel, or as a mixture in any concentration with regular petroleum diesel or other biodiesel.

El biodiesel ofrece diversas propiedades beneficiosas en comparacion con el diesel basado en petroleo, incluyendo emisiones reducidas (por ejemplo, monoxido de carbono, azufre, hidrocarburos aromaticos, particulas de hollin, etc.) durante la combustion. El biodiesel tambien mantiene un ciclo equilibrado de dioxido de carbono porque se basa en materiales biologicos renovables. Normalmente el biodiesel es completamente biodegradable y tiene un buen perfil de seguridad debido a su punto de ignicion relativamente alto y a su baja combustibilidad. Asimismo, el biodiesel proporciona buenas propiedades lubricantes, reduciendo asi el desgaste y la rotura de los motores.

Biodiesel offers several beneficial properties compared to petroleum-based diesel, including reduced emissions (for example, carbon monoxide, sulfur, aromatic hydrocarbons, soot particles, etc.) during combustion. Biodiesel also maintains a balanced cycle of carbon dioxide because it is based on renewable biological materials. Normally, biodiesel is completely biodegradable and has a good safety profile due to its relatively high ignition point and low combustibility. Also, biodiesel provides good lubricating properties, thus reducing engine wear and tear.

Los metodos actuales para producir biodiesel implican la transesterificacion de triacilgliceridos a partir de materias primas de aceites vegetales, tales como de la colza en Europa, de la soja en norteamerica y de aceite de palma en el sudeste asiatico. Por lo tanto, la produccion de biodiesel a escala industrial esta restringida geografica y estacionalmente a zonas donde se producen materias primas de aceites vegetales. El proceso de transesterificacion conduce a una mezcla de esteres grasos que puede utilizarse como biodiesel, aunque tambien a un subproducto indeseable, la glicerina. Para ser utilizables como biodiesel, los esteres grasos deben purificarse adicionalmente del producto heterogeneo. Esto aumenta los costes y la cantidad de energia requerida para la produccion de esteres grasos y, finalmente, tambien para la produccion de biodiesel. Asimismo, las materias primas de aceite vegetal son fuentes de energia ineficaces porque requieren una extensa superficie de cultivo. Por ejemplo, el rendimiento de biodiesel procedente de la colza es de solo 1300 l/hectarea porque solo se utiliza el aceite de la semilla para la produccion de biodiesel, y no el resto de la biomasa de la colza. Adicionalmente, cultivar algunas materias primas de aceite vegetal, como la colza y la soja, requiere una rotacion frecuente del cultivo para evitar el empobrecimiento de nutrientes de la tierra.

Current methods for producing biodiesel involve the transesterification of triacylglycerides from raw materials of vegetable oils, such as rapeseed in Europe, soybeans in North America and palm oil in Southeast Asia. Therefore, the production of biodiesel on an industrial scale is geographically and seasonally restricted to areas where raw materials of vegetable oils are produced. The transesterification process leads to a mixture of fatty esters that can be used as biodiesel, but also to an undesirable by-product, glycerin. To be usable as biodiesel, fatty esters must be further purified from the heterogeneous product. This increases the costs and the amount of energy required for the production of fatty esters and, finally, also for the production of biodiesel. Likewise, vegetable oil raw materials are inefficient sources of energy because they require an extensive cultivation area. For example, the biodiesel yield from rapeseed is only 1300 l / hectare because only the seed oil is used for biodiesel production, and not the rest of the rapeseed biomass. Additionally, growing some vegetable oil raw materials, such as rapeseed and soybeans, requires frequent crop rotation to avoid depletion of nutrients from the soil.

La publicacion PCT No. WO 2007/136762 desvela microorganismos recombinantes que son capaces de sintetizar productos procedentes de la ruta sintetica de acidos grasos, incluyendo, entre otros, esteres de acidos grasos y alcoholes grasos. En particular, se describen determinados derivados de acidos grasos que tienen una longitud, una ramificacion y niveles de saturacion definidos de la cadena de carbonos. La publicacion '762 describe celulas recombinantes que utilizan sobreexpresion endogena o expresion heterologa de proteinas tioesterasas en la produccion de derivados de acidos grasos.

PCT Publication No. WO 2007/136762 discloses recombinant microorganisms that are capable of synthesizing products from the synthetic fatty acid pathway, including, among others, fatty acid esters and fatty alcohols. In particular, certain fatty acid derivatives having a defined length, branching and saturation levels of the carbon chain are described. Publication '762 describes recombinant cells that use endogenous overexpression or heterologous expression of thioesterase proteins in the production of fatty acid derivatives.

La publicacion PCT No. WO 2008/119082 desvela celulas y microorganismos genomanipulados que producen productos a partir de la ruta biosintetica de acidos grasos, incluyendo, entre otros, esteres de acidos grasos y alcoholes grasos. La publicacion '082 describe celulas recombinantes que utilizan la sobreexpresion de enzimas acil-CoA sintetasa para producir de manera mas eficiente derivados de acidos grasos.

PCT Publication No. WO 2008/119082 discloses genomanipulated cells and microorganisms that produce products from the biosynthetic pathway of fatty acids, including, among others , esters of fatty acids and fatty alcohols. Publication '082 describes recombinant cells that use overexpression of acyl-CoA synthetase enzymes to more efficiently produce fatty acid derivatives.

La Patente de Estados Unidos N° 5.955.329 desvela proteinas acil-ACP tioesterasas de plantas genomanipuladas (manipuladas mediante ingenieria genetica), que tienen especificidad de sustrato alterada. En particular, la patente '329 desvela la produccion de acil-ACP tioesterasas de plantas genomanipuladas, en donde las acil-ACP tioesterasas de plantas genomanipuladas demuestran especificidad de sustrato alterada con respecto a sustratos de acil-ACP hidrolizados por las tioesterasas de plantas en comparacion con la acil-ACP tioesterasa nativa.

US Patent No. 5,955,329 discloses acyl-ACP thioesterase proteins from genomanipulated plants (engineered by genetic engineering), which have altered substrate specificity. In particular, the '329 patent discloses the production of acyl-ACP thioesterase from genomanipulated plants, wherein the acyl-ACP thioesterase from genomanipulated plants demonstrate altered substrate specificity with respect to substrates of acyl-ACP hydrolyzed by plant thiosterases compared to native acyl-ACP thioesterase.

Lee et al. (Biochimica et Biophysica Acta 1774 (2007) 959-967) describen una variante de la tioesterasa de SEC ID NO: 31 que tiene una sustitucion de aminoacido en la posicion 109 (L109P), conservando dicha variante solo aproximadamente el 5 % de las actividades de la enzima nativa hacia tanto la lauroil como la estearoil-CoA.

Lee et al. (Biochimica et Biophysica Acta 1774 (2007) 959-967) describe a thioesterase variant of SEQ ID NO: 31 that has an amino acid substitution at position 109 (L109P), with said variant retaining only about 5% of the activities from the native enzyme to both lauroyl and stearoyl-CoA.

Mientras que la tecnica anterior desvela ciertas descripciones utiles con respecto a la produccion de ciertos derivados de acidos grasos, existe una necesidad en el campo de metodos y procedimientos mejorados para una produccion mas eficaz y economica de dichos derivados de acidos grasos, y tambien para tecnologia que facilite la produccion de composiciones que tengan especificaciones de producto alteradas. Como ejemplo especifico, existe una necesidad para la produccion de composiciones de acidos grasos que tengan especificaciones y propiedades previamente disenadas o "adaptadas" para aplicaciones particulares tales como combustibles, detergentes, lubricantes, moleculas precursoras industriales y otras aplicaciones valiosas de derivados de acidos grasos.

While the prior art discloses certain useful descriptions regarding the production of certain fatty acid derivatives, there is a need in the field of improved methods and procedures for more efficient and economical production of said fatty acid derivatives, and also for technology. that facilitates the production of compositions that have altered product specifications. As a specific example, there is a need for the production of fatty acid compositions having specifications and properties previously designed or "adapted" for particular applications such as fuels, detergents, lubricants, industrial precursor molecules and other valuable applications of fatty acid derivatives.

Sumario de la invencion

Summary of the invention

La presente divulgacion se refiere a enzimas tioesterasas mutantes y naturales utiles, a polinucleotidos que codifican estas enzimas, a vectores que comprenden los polinucleotidos que codifican las enzimas tioesterasas utiles, a celulas hospedadoras recombinantes que comprenden enzimas tioesterasas endogenas mutadas, a celulas hospedadoras recombinantes transformadas con los vectores, a celulas hospedadoras recombinantes que tienen polinucleotidos que codifican enzimas tioesterasas utiles cromosomicamente integradas en ellas, a tioesterasas producidas por las celulas hospedadoras, a composiciones derivadas de acidos grasos (tales como productos quimicos industriales y biocombustibles) producidas in vitro y/o in vivo, a metodos para producir composiciones derivadas de acidos grasos in vitro y/o in vivo, y a metodos de uso de las composiciones producidas derivadas de acidos grasos.

The present disclosure relates to useful mutant and natural thioesterase enzymes, to polynucleotides encoding these enzymes, to vectors comprising polynucleotides that encode useful thioesterase enzymes, to recombinant host cells comprising mutated endogenous thioesterase enzymes, to recombinant host cells transformed with the vectors, to recombinant host cells having polynucleotides encoding chromosomally useful thioesterase enzymes integrated therein, to thioesterase produced by the host cells, to fatty acid derived compositions (such as industrial chemicals and biofuels) produced in vitro and / or in vivo, to methods for producing compositions derived from fatty acids in vitro and / or in vivo, and to methods of using the compositions produced derived from fatty acids.

La presente divulgacion se refiere a metodos para producir composiciones derivadas de acidos grasos a traves de fermentaciones microbianas que tienen perfiles de productos predeterminados con respecto a las longitudes de las cadenas de carbono y rendimientos proporcionales. Estas composiciones son muy adecuadas para aplicaciones en las industrias quimica y de combustibles porque sus propiedades pueden adaptarse a las aplicaciones particulares para las que estan destinadas. Por ejemplo, es posible adaptar un producto de ester graso, segun los metodos descritos en el presente documento, de tal manera que pueda utilizarse como combustible para automoviles y/o para disenar una composicion que tenga, por ejemplo, caracteristicas de combustible mejoradas, tales como el punto de enturbamiento, lubricidad, numero de cetano, viscosidad cinematica, numero de acido, punto de ebullicion, estabilidad oxidativa, punto de obstruccion del filtro en frio, perfil de impurezas, nivel de ceniza sulfatada y/o punto de ignicion. Del mismo modo, es posible producir productos quimicos industriales segun los metodos descritos en el presente documento que puedan reemplazar a los productos quimicos actuales procedentes de fuentes de petroleo, y que esten adaptados a aplicaciones particulares, por ejemplo, para producir alcoholes grasos que son optimamente idoneos para su uso como tensioactivos y/o detergentes.

The present disclosure relates to methods for producing compositions derived from fatty acids through microbial fermentations having predetermined product profiles with respect to carbon chain lengths and proportional yields. These compositions are very suitable for applications in the chemical and fuel industries because their properties can be adapted to the particular applications for which they are intended. For example, it is possible to adapt a fatty ester product, according to the methods described herein, so that it can be used as automobile fuel and / or to design a composition having, for example, improved fuel characteristics, such such as cloud point, lubricity, cetane number, kinematic viscosity, acid number, boiling point, oxidative stability, cold filter clogging point, impurity profile, sulfated ash level and / or ignition point. Similarly, it is possible to produce industrial chemicals according to the methods described herein that can replace current chemicals from petroleum sources, and that are adapted to particular applications, for example, to produce fatty alcohols that are optimally suitable for use as surfactants and / or detergents.

La presente divulgacion se refiere a metodos alternativos para producir esteres grasos sin la presencia (o en ausencia) de una ester sintasa. Este metodo es energeticamente mas favorable que los metodos hasta ahora desvelados para producir composiciones de esteres grasos a traves de procesos de fermentacion microbiana, que requieren al menos tanto una enzima tioesterasa como una enzima ester sintasa. Como tales, las nuevas tioesterasas de la divulgacion proporcionan ventajas adicionales. La presente invencion esta definida por las reivindicaciones. En particular, la presente invencion proporciona una enzima tioesterasa genomanipulada que transforma un sustrato de acil-ACP C10, C12 o C14 en un derivado de acido graso con una mayor actividad en comparacion con la tioesterasa de tipo silvestre que tiene la secuencia de aminoacidos presentada como SEC ID NO: 31, en donde la tioesterasa genomanipulada (a) procede de una tioesterasa precursora que comprende una secuencia de aminoacidos que es al menos 90% identica a la SEC ID NO: 31, y (b) tiene una sustitucion correspondiente a una sustitucion seleccionada de:

The present disclosure relates to alternative methods for producing fatty esters without the presence (or absence) of an ester synthase. This method is energetically more favorable than the methods so far disclosed to produce fatty ester compositions through microbial fermentation processes, which require at least both a thioesterase enzyme and an ester synthase enzyme. As such, the new thioesterase of the disclosure provide additional advantages. The present invention is defined by the claims. In particular, the present invention provides a genomanipulated thioesterase enzyme that transforms a C10, C12 or C14 acyl-ACP substrate into a fatty acid derivative with increased activity compared to the wild-type thioesterase having the amino acid sequence presented as SEQ ID NO: 31, wherein the genomanipulated thioesterase (a) is derived from a precursor thioesterase comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 31, and (b) has a substitution corresponding to a Selected substitution of:

i) D45A, D45C, D45E, D45F, D45G, D45H, D45I, D45K, D45L, D45M, D45P, D45Q, D45S, D45T, D45V, D45W; ii) N73A, N73C, N73G, N73H, N73I, N73L, N73P, N73R, N73S, N73T, N73V, N73W;

i) D45A, D45C, D45E, D45F, D45G, D45H, D45I, D45K, D45L, D45M, D45P, D45Q, D45S, D45T, D45V, D45W; ii) N73A, N73C, N73G, N73H, N73I, N73L, N73P, N73R, N73S, N73T, N73V, N73W;

iii) G78A, G78C, G78D, G78E, G78F, G78M, G78N, G78P, G78Q, G78R, G78S, G78T, G78V, G78Y;

iii) G78A, G78C, G78D, G78E, G78F, G78M, G78N, G78P, G78Q, G78R, G78S, G78T, G78V, G78Y;

iv) Q80A, Q80E, Q80G, Q80L, Q80M, Q80S, Q80W, Q80Y;

iv) Q80A, Q80E, Q80G, Q80L, Q80M, Q80S, Q80W, Q80Y;

v) E101A, E101D, E101G, E101L, E101M, E101P, E101S, E101T, E101V;

v) E101A, E101D, E101G, E101L, E101M, E101P, E101S, E101T, E101V;

vi) R108A, R108C, R108D, R108E, R108F, R108G, R108H, R108I, R108L, R108M, R108R, R108S, R108V, R108W, R108Y;

vi) R108A, R108C, R108D, R108E, R108F, R108G, R108H, R108I, R108L, R108M, R108R, R108S, R108V, R108W, R108Y;

vii) A111C, A111E, A111L, A111M, A111N, A111P, A111Q, A111R, A111S, A111V, A111W, A111Y;

vii) A111C, A111E, A111L, A111M, A111N, A111P, A111Q, A111R, A111S, A111V, A111W, A111Y;

viii) Y117A, Y117C, Y117D, Y117E, Y117G, Y117H, Y117I, Y117L, Y117M, Y117N, Y117P, Y117Q, Y117R, Y117S, Y117T, Y117V, Y117W;

viii) Y117A, Y117C, Y117D, Y117E, Y117G, Y117H, Y117I, Y117L, Y117M, Y117N, Y117P, Y117Q, Y117R, Y117S, Y117T, Y117V, Y117W;

ix) N118A, N118C, N118E, N118F, N118G, N118H, N118I, N118K, N118L, N118M, N118P, N118Q, N118S, N118T, N118V, N118W;

ix) N118A, N118C, N118E, N118F, N118G, N118H, N118I, N118K, N118L, N118M, N118P, N118Q, N118S, N118T, N118V, N118W;

x) S122A, S122C, S122D, S122E, S122F, S122G, S122I, S122L, S122M, S122P, S122R, S122V, S122W, S122Y;

x) S122A, S122C, S122D, S122E, S122F, S122G, S122I, S122L, S122M, S122P, S122R, S122V, S122W, S122Y;

xi) Y145A, Y145C, Y145D, Y145E, Y145G, Y145I, Y145L, Y145M, Y145N, Y145Q, Y145R, Y145S, Y145T, Y145W;

xi) Y145A, Y145C, Y145D, Y145E, Y145G, Y145I, Y145L, Y145M, Y145N, Y145Q, Y145R, Y145S, Y145T, Y145W;

xii) Q152A, Q152D, Q152E, Q152F, Q152H, Q152I, Q152K, Q152L, Q152N, Q152R, Q152S, Q152T, Q152V, Q152Y;

xii) Q152A, Q152D, Q152E, Q152F, Q152H, Q152I, Q152K, Q152L, Q152N, Q152R, Q152S, Q152T, Q152V, Q152Y;

xiii) V178A, V178F, V178G, V178L, V178R, V178S, V178T, V178W;

xiii) V178A, V178F, V178G, V178L, V178R, V178S, V178T, V178W;

de la SEC ID NO: 31, en donde la metionina esta en la posicion 0 y alanina esta en la posicion 1 de la SEC ID NO: 31 (Fig. 58).

of SEQ ID NO: 31, wherein the methionine is in position 0 and alanine is in position 1 of SEQ ID NO: 31 (Fig. 58).

Se desvelan tioesterasas mutantes (o sus equivalentes naturales) que proceden de una tioesterasa precursora, en donde cada una de las mutantes (o los equivalentes naturales) tiene al menos una propiedad alterada in vitro y/o in vivo, en comparacion con las propiedades de la tioesterasa precursora. La propiedad alterada puede ser, por ejemplo, una propiedad biofisica, tal como estabilidad termica (punto de fusion T^m (melting, fusion)); estabilidad de disolventes, solutos y/u oxidativa; lipofilicidad; hidrofilidad; estructura cuaternaria; momento dipolar; y/o punto isoelectrico. La propiedad alterada tambien puede ser, por ejemplo, una propiedad bioquimica, tal como pH optimo, temperatura optima y/o fuerza ionica optima. Adicionalmente, la propiedad alterada puede ser, por ejemplo, un parametro catalitico enzimatico, tal como la distribucion del producto (incluyendo, por ejemplo, un mayor o menos porcentaje o rendimiento proporcional para un producto en particular en comparacion con otros productos en la mezcla del producto), actividad especifica, preferencia de sustrato, afinidad de sustrato, inhibicion del sustrato, afinidad del producto, velocidad de recambio o velocidad catalitica, inhibicion del producto, mecanismo cinetico, K^m, k^cat, k^cat/K^m y/o V^Max. Adicionalmente, la propiedad alterada es, por ejemplo, un aumento o una disminucion de la actividad o un cambio en la preferencia por la alcoholisis frente a la hidrolisis, sustratos de acil proteina transportadora de acil-CoA frente a acil proteina transportadora de acilos, sustratos de esteres frente a tioester, sustratos saturados frente a insaturados, sustratos de cadena lineal frente a ramificados; cambios en las posiciones de insaturacion, intervalos de numeros de cetano, o longitudes de cadenas de carbono especificas, sustratos ramificados, posicion de ramificacion, sustratos de hidroxiacilo, sustratos de ceto-acilo; y/o productos con un intervalo cambiado de numeros de cetano o con numeros de cetano especificos, indice de octano, estabilidad oxidativa, lubricidad, punto de ignicion, viscosidad, punto de ebullicion, punto de fusion, punto de fluidez, punto de enturbamiento, punto de obstruccion del filtro en frio, caracteristicas de flujo en frio, aromaticidad y/o numero de yodos. Las propiedades alteradas tambien pueden incluir, por ejemplo, una disminucion de la actividad o una atenuacion de la hidrolisis de los esteres, de tal manera que la hidrolisis de las moleculas del producto deseado se reduce o elimina. Las propiedades alteradas pueden incluir, ademas, por ejemplo, una disminucion en la toxicidad de la proteina para la celula y/o un cambio en el nivel de expresion de la proteina en la celula, en comparacion con la toxicidad de la proteina precursora para la celula y/o con el nivel de expresion en la misma celula. En un aspecto ejemplar, una propiedad alterada puede incluir un cambio en la capacidad para catalizar la sintesis de derivados de acilo graso directa o indirectamente in vivo o in vitro. En otra realizacion ejemplar, una propiedad alterada es la mejora o aumento del rendimiento in vitro y/o in vivo o rendimiento proporcional de un derivado de acido graso particularmente deseable.

Mutant thiosterases (or their natural equivalents) that are derived from a precursor thioesterase are disclosed, wherein each of the mutants (or natural equivalents) has at least one property altered in vitro and / or in vivo, as compared to the properties of the thioesterase precursor. The altered property can be, for example, a biophysical property, such as thermal stability (melting point T ^m ( melting, melting)); solvent, solute and / or oxidative stability; lipophilicity; hydrophilicity; quaternary structure; dipole moment; and / or isoelectric point. The altered property can also be, for example, a biochemical property, such as optimum pH, optimum temperature and / or optimal ionic strength. Additionally, the altered property may be, for example, an enzymatic catalytic parameter, such as product distribution (including, for example, a greater or lesser percentage or proportional yield for a particular product compared to other products in the mixture of the product), specific activity, substrate preference, substrate affinity, substrate inhibition, product affinity, turnover rate or catalytic speed, product inhibition, kinetic mechanism, K ^m , k ^cat , k ^cat / K ^m and / or V ^Max . Additionally, the altered property is, for example, an increase or decrease in activity or a change in the preference for alcohololysis versus hydrolysis, acyl-AcA transporter protein substrates over acyl-acyl transporter protein, substrates of esters against thioester, saturated versus unsaturated substrates, straight chain versus branched substrates; changes in unsaturation positions, cetane number ranges, or specific carbon chain lengths, branched substrates, branching position, hydroxyacyl substrates, keto-acyl substrates; and / or products with a changed range of cetane numbers or with specific cetane numbers, octane number, oxidative stability, lubricity, ignition point, viscosity, boiling point, melting point, pour point, cloud point, cold filter clogging point, cold flow characteristics, aromaticity and / or number of iodines. The altered properties may also include, for example, a decrease in activity or an attenuation of the hydrolysis of the esters, such that the hydrolysis of the molecules of the desired product is reduced or eliminated. The altered properties may also include, for example, a decrease in the toxicity of the protein for the cell and / or a change in the level of expression of the protein in the cell, as compared to the toxicity of the precursor protein for the cell. cell and / or with the level of expression in the same cell. In an exemplary aspect, an altered property may include a change in the ability to catalyze the synthesis of fatty acyl derivatives directly or indirectly in vivo or in vitro. In another exemplary embodiment, an altered property is the improvement or increase of the in vitro and / or in vivo yield or proportional yield of a particularly desirable fatty acid derivative.

En un aspecto, una tioesterasa mutante (o uno de sus equivalentes naturales) procede de una tioesterasa precursora. En un aspecto particular, la tioesterasa precursora es una tioesterasa natural, una tioesterasa previamente modificada, o una tioesterasa sintetica.

In one aspect, a mutant thioesterase (or one of its natural equivalents) comes from a precursor thioesterase. In a particular aspect, the precursor thioesterase is a natural thioesterase, a previously modified thioesterase, or a synthetic thioesterase.

En un aspecto, la tioesterasa mutante (o uno de sus equivalentes naturales) procede de una tioesterasa precursora que es una tioesterasa natural. La tioesterasa precursora natural puede obtenerse a partir de, por ejemplo, una planta, un animal, una bacteria, un hongo, una levadura, o de otras fuentes microbianas. La tioesterasa mutante (o uno de sus equivalentes naturales) puede proceder de una tioesterasa precursora de los siguientes generos Acidovorax, Acinetobacter, Aeromonas, Alcanivorax, Aliivibrio, Alkalilimnicola, Alteromonadales, Alteromonas, Aurantimonas, Azoarcus, Azorhizobium, Azotobacter, Beggiatoa, Beijerinckia, Bordetella, Bradyrhizobium, Burkholderia, Caulobacter, Cellvibrio, Chromobacterium, Citrobacter, Comamonas, Cupriavidus, Dechloromonas, Delftia, Desulfovibrio, Enterobacter, Erwinia, Escherichia, Geobacter, Hahella, Halorhodospira, Herminiimonas, Idiomarina, Janthinobacterium, Klebsiella, Leptospira, Leptotrix, Limnobacter, Magnetospirillum, Marinobacter, Marinomonas, Metilibio, Metilobacilo, Methylobacterium, Metilocella, Methylococcus, Moritella, Nitrobacter, Nitrococcus, Nitrosomonas, Nitrosospira, Oceanospirillum, Oligotropha, Pectobacterium, Photobacterium, Photorhabdus, Polaromonas, Proteus, Providencia, Pseudoalteromonas, Pseudomonas, Psychromonas, Ralstonia, Reinekea, Rhodobacterales, Rhodoferax, Rhodopseudomonas, Rhodospirillum, Saccharophagus, Salmonella, Serratia, Shewanella, Shigella, Stenotrophomonas, Streptococcus, Thauera, Thioalkalivibrio, Thiobacillus, Vibrio, Xanthomonas o Yersinia.

In one aspect, the mutant thioesterase (or one of its natural equivalents) is derived from a precursor thioesterase which is a natural thioesterase. Natural precursor thioesterase can be obtained from, for example, a plant, an animal, a bacterium, a fungus, a yeast, or other microbial sources. The mutant thioesterase (or one of its natural equivalents) may be derived from a precursor thioesterase of the following genera Acidovorax, Acinetobacter, Aeromonas, Alcanivorax, Aliivibrio, Alkalilimnicola, Alteromonadales, Alteromonas, Aurantimonas, Azoarcus, Azorhizobacteria, Azorhizobacteria, Azorhizobacteria, Azorhizobacteria, Azorhizobacteria, Azorhizobacteria, Azorhizobacteria, Azorhizobacteria, Azorhizobacteria, Azorhizobacteria, Azorhizobacteria, Azorhizobacteria, Azorhizobacteria, Azorhizobacteria , Bradyrhizobium, Burkholderia, Caulobacter, Cellvibrio, Chromobacterium, Citrobacter, Comamonas, Cupriavidus, Dechloromonas, Delftia, Desulfovibrio, Enterobacter, Erwinia, Escherichia, Geobacter, Hahella, Halorhodospira, Herminiimonas, Idiomarina, Janthinobacterium, Klebsiella, Leptospira, Leptotrix, Limnobacter, Magnetospirillum , Marinobacter, Marinomonas, Metilibio, Metilobacilo, Methylobacterium, Metilocella, Methylococcus, Moritella, Nitrobacter, Nitrococcus, Nitrosomonas, Nitrosospira, Oceanospirillum, Oligotropha, Pectobacterium, Photobacterium, Photorhabdus, Polaromonas, Proteus, Providencia, Pseudoalteromonas, Pseudomonas, Psychromonas, Ralstonia, Reinek ea, Rhodobacterales, Rhodoferax, Rhodopseudomonas, Rhodospirillum, Saccharophagus, Salmonella, Serratia, Shewanella, Shigella, Stenotrophomonas, Streptococcus, Thauera, Thioalkalivibrio, Thiobacillus, Vibrio, Xanthomonas or Yersinia.

En un aspecto particular, la tioesterasa precursora puede proceder de cualquiera de Acidovorax avenae subsp. citrulli AAC00-1, Acidovorax sp. JS42, Acinetobacter baumannii ACICU, Acinetobacter baumannii ATCC 17978, Aeromonas hydrophila subsp. Hydrophila ATCC 7966, Aeromonas salmonicida subsp. salmonicida A449, Alcanivorax borkumensis SK2, Alcanivorax sp. DG881, Aliivibrio salmonicida LFI1238, Alkalilimnicola ehrlichei MLHE-1, alfa proteobacterium HTCC2255, Alteromonadales bacterium TW-7, ecotipo profundo de Alteromonas macleodii, Aurantimonas sp. SI85-9A1, Azoarcus sp. BH72, Azorhizobium caulinodans ORS 571, Azotobacter vinelandii AvOP, Beggiatoa sp. PS, Beijerinckia indica subsp. indica ATCC 9039, Bordetella avium 197N, Bordetella bronchiseptica RB50, Bordetella parapertussis 12822, Bordetella pertussis Tohama I, Bordetella petrii DSM 12804, Bradyrhizobium sp. BTAil, Bradyrhizobium sp. ORS278, Burkholderia ambifaria AMMD, Burkholderia ambifaria IOP40-10, Burkholderia ambifaria MC40-6, Burkholderia ambifaria MEX-5, Burkholderia cenocepacia AU 1054, Burkholderia cenocepacia HI2424, Burkholderia cenocepacia J2315, Burkholderia cenocepacia MC0-3, Burkholderia cenocepacia PC184, Burkholderia dolosa AUO158, Burkholderia graminis C4D1M, Burkholderia mallei ATCC 23344, Burkholderia mallei GB8 horse 4, Burkholderia mallei NCTC 10229, Burkholderia multivorans ATCC 17616, Burkholderia oklahomensis C6786, Burkholderia oklahomensis EO147, Burkholderia phymatum STM815, Burkholderia pseudomallei 1106a, Burkholderia pseudomallei 1106b, Burkholderia pseudomallei 14, Burkholderia pseudomallei 1655, Burkholderia pseudomallei 1710b, Burkholderia pseudomallei 305, Burkholderia pseudomallei 406e, Burkholderia pseudomallei 668, Burkholderia pseudomallei 7894, Burkholderia pseudomallei K96243, Burkholderia pseudomallei NCTC 13177, Burkholderia sp. 383, Burkholderia thailandensis Bt4, Burkholderia thailandensis E264, Burkholderia thailandensis MSMB43, Burkholderia thailandensis TXDOH, Burkholderia ubonensis Bu, Burkholderia vietnamiensis G4, Caulobacter crescentus CB15, Cellvibrio japonicus Ueda107, Chromobacterium violaceum ATCC 12472, Chromohalobacter salexigens DSM 3043, Citrobacter koseri ATCC BAA-895, Comamonas testosteroni KF-1, Cupriavidus taiwanensis, Dechloromonas aromatica RCB, Delftia acidovorans SPH-1, Desulfovibrio desulfuricans subsp. desulfuricans cepa G20, Desulfovibrio desulfuricans subsp. desulfuricans cepa G20, Enterobacter cancerogenus ATCC 35316, Enterobacter sakazakii ATCC BAA-894, Enterobacter sp. 638, Erwinia tasmaniensis, Escherichia albertii TW07627, Escherichia coli O157.H7 EDL933, Escherichia coli O157:H7 cepa EC4024, Escherichia coli O157:H7 cepa EC4196, gamma proteobacterium HTCC5015, gamma proteobacterium KT 71, Geobacter sp. M21, Hahella chejuensis KCTC 2396, Halorhodospira halophila SL1, Herminiimonas arsenicoxydans, Idiomarina baltica OS145, Idiomarina loihiensis L2TR, Janthinobacterium sp. Marseille, Klebsiella pneumoniae 342, Klebsiella pneumoniae subsp. pneumoniae MGH 78578, Klebsiella sp. ZD414, Leptospira interrogans serovar Copenhageni cepa Fiocruz L1-130, Leptospira interrogans serovar Copenhageni cepa 56601, Leptothrix cholodnii SP-6, Limnobacter sp. MED105, Magnetospirillum magneticum AMB-1, gamma proteobacterium marina HTCC2080, gamma proteobacterium marina HTCC2143, gamma proteobacterium marina HTCC2207, metagenoma marino, Marinobacter algicola DG893, Marinobacter aquaeolei VT8, Marinobacter sp. ELB17, Marinomonas sp. MWYL1, Methylibium petroleiphilum PM1, Methylobacillus flagellatus KT, Methylobacterium chloromethanicum CM4, Methylobacterium extorquens PA1, Methylobacterium populi BJ001, Methylocella silvestris BL2, Methylococcus capsulatus cepa Bath, Moritella sp. PE36, Nitrobacter sp. Nb-311A, Nitrobacter winogradskyi Nb-255, Nitrococcus mobilis nb-231, Nitrosococcusoceani ATCC 19707, Nitrosococcus oceani C-27, Nitrosomonas europaea ATCC 19718, Nitrosomonas eutropha C91, Nitrosospira multiformis ATCC 25196, Oceanospirillum sp. MED92, Oligotropha carboxidovorans OM5, Pectobacterium atrosepticum SCRI1043, Photobacterium profundum 3TCK, Photobacterium profundum SS9, Photobacterium sp. SKA34, Photorhabdus luminescens, Photorhabdus luminescens subsp. laumondii TTO1, Polaromonas naftalenivoranos CJ2, Polaromonas sp. JS666, Polynucleobacter sp. QLW-P1DMWA-1, Proteus mirabilis HI4320, Providencia stuartii ATCC 25827, Pseudoalteromonas atlantica T6c, Pseudoalteromonas haloplanktis TAC125, Pseudoalteromonas sp. 643A, Pseudoalteromonas tunicata D2, Pseudomonas aeruginosa PA7, Pseudomonas aeruginosa PACS2, Pseudomonas aeruginosa PAO1, Pseudomonas aeruginosa UCBPP-PA14, Pseudomonas entomophila L48, Pseudomonas fluorescens Pf0-1, Pseudomonas fluorescens Pf-5, Pseudomonas mendocina ymp, Pseudomonas putida F1, Pseudomonas putida GB-1, Pseudomonas putida KT2440, Pseudomonas putida W619, Pseudomonas stutzeri A1501, Pseudomonas syringae pv. Phaseolicola 1448A, Pseudomonas syringae pv. syringae B728a, Pseudomonas syringae pv. tomate cepa DC3000, Psychromonas ingrahamii 37, Ralstonia eutropha H16, Ralstonia eutropha JMP134, Ralstonia metallidurans CH34, Ralstonia pickettii 12D, Ralstonia pickettii 12J, Ralstonia solanacearum GMI1000, Ralstonia solanacearum IPO1609, Ralstonia solanacearum MolK2, Ralstonia solanacearum UW551, Reinekea sp. Med297, Bacteria Rhodobacterales Y4I, Rhodoferax ferrireducens T118, Rhodopseudomona spalustris BisA53, Rhodopseudomonas palustris BisB18, Rhodopseudomonas palustris BisB5, Rhodopseudomonas palustris CGA009, Rhodopseudomonas palustris HaA2, Rhodopseudomonas palustris TIE-1, Rhodospirillum centenum SW, Saccharophagus degradans 2-40, Salmonella enterica subsp. arizonae serovar 62:z4,z23:--,Salmonella enterica subsp. enterica serovar Choleraesuis cepa SC-B67, Salmonella enterica subsp. Enterica serovar allinarum cepa 287/91, Salmonella enterica subsp. Enterica serovar Hadar cepa RI_05P066, Salmonella enterica subsp. enterica serovar Javiana cepa GA_MM04042433, Salmonella enterica subsp. enterica serovar Saintpaul cepa SARA23, Salmonella enterica subsp. enterica serovar Saintpaul cepa SARA29, Salmonella enterica subsp. Enterica serovar Typhi cepa CT18, Salmonella typhimurium LT2, Serratia proteamaculans 568, Shewanella amazonensis SB2B, Shewanella baltica OS155, Shewanella baltica OS185, Shewanella baltica OS195, Shewanella baltica OS223, Shewanella benthica KT99, Shewanella denitrificans OS217, Shewanella frigidimarina NCIMB 400, Shewanella halifaxensis HAW-EB4, Shewanella loihica PV-4, Shewanella oneidensis MR-1, Shewanella pealeana ATCC 700345, Shewanella putrefaciens 200, Shewanella sediminis HAW-EB3, Shewanella sp. ANA-3, Shewanella sp. MR-4, Shewanella sp. MR-7, Shewanella sp. W3-18-1, Shewanella woodyi ATCC 51908, Shigella boydii Sb227, Shigella dysenteriae Sd197, Stenotrophomonas maltophilia K279a, Stenotrophomonas maltophilia R551-3, Streptococcus sp. (N1), construccion sintetica, Thauera sp. MZ1T, Thioalkalivibrio sp. HL-EbGR7, Thiobacillus denitrificans ATCC25259, Thiomicrospira crunogena x CL-2, Vibrio alginolyticus 12G01, Vibrio angustum S14, Vibrio campbellii AND4, Vibrio cholerae 2740-80, Vibrio cholerae MZO-2, Vibrio cholerae O1 biovar eltor cepa N16961, Vibrio cholerae V51, Vibrio fischeri ES114, Vibrio fischeri MJ11, Vibrio harveyi ATCC BAA-1116, Vibrio mimicus, Vibrionales bacterium SWAT-3, Vibrio parahaemolyticus AQ3810, Vibrio parahaemolyticus RIMD 2210633, Vibrio shilonii AK1, Vibrio splendidus 12B01, Vibrio sp. MED222, Vibrio vulnificus CMCP6, Vibrio vulnificus YJ016, Xanthomonas axonopodis pv. citri cepa 306, Xanthomonas campestris pv. campestris cepa ATCC 33913, Xanthomonas campestris pv. campestris cepa B100, Xanthomonas campestris pv. Vesicatoria cepa 85-10, Xanthomonas oryzae pv. oryzae KACC10331, Xanthomonas oryzae pv. oryzae PXO99A, Xanthomonas oryzae pv. oryzicola BLS256, Yersinia bercovieri ATCC 43970, Yersinia enterocolitica subsp. enterocolitica 8081, Yersinia frederiksenii ATCC 33641, Yersinia intermedia ATCC 29909, Yersinia mollaretii ATCC 43969, Yersinia pestis Angola, Yersinia pestis biovar Orientalis cepa F1991016, Yersinia pestis CO92, Yersinia pestis KIM o Yersinia pseudotuberculosis IP 31758.

In a particular aspect, the precursor thioesterase can be derived from any of Acidovorax avenae subsp. citrulli AAC00-1, Acidovorax sp. JS42, Acinetobacter baumannii ACICU, Acinetobacter baumannii ATCC 17978, Aeromonas hydrophila subsp. Hydrophila ATCC 7966, Aeromonas salmonicida subsp. Salmonicide A449, Alcanivorax borkumensis SK2, Alcanivorax sp. DG881, Salmonicide antibiotic LFI1238, Alkalilimnicola ehrlichei MLHE-1, alpha proteobacterium HTCC2255, Alteromonadales bacterium TW-7, deep ecotype of Alteromonas macleodii, Aurantimonas sp. SI85-9A1, Azoarcus sp. BH72, Azorhizobium caulinodans ORS 571, Azotobacter vinelandii AvOP, Beggiatoa sp. PS, Beijerinckia indica subsp. Indicates ATCC 9039, Bordetella avium 197N, Bordetella bronchiseptica RB50, Bordetella parapertussis 12822, Bordetella pertussis Tohama I, Bordetella petrii DSM 12804, Bradyrhizobium sp. BTAil, Bradyrhizobium sp. ORS278, Burkholderia ambifaria AMMD, Burkholderia ambifaria IOP40-10, Burkholderia ambifaria MC40-6, Burkholderia ambifaria MEX-5, Burkholderia cenocepacia AU 1054, Burkholderia cenocepacia HI2424, Burkholderia cenocepacia J2315, Burkholderia cenoceo08 , Burkholderia graminis C4D1M, Burkholderia mallei ATCC 23344, Burkholderia mallei GB8 horse 4, Burkholderia mallei NCTC 10229, Burkholderia multivorans ATCC 17616, Burkholderia oklahomensis C6786, Burkholderia oklahomensis EO147, Burkholderia pymatumia, 6th pseudo Burkholderia pseudomallei 1655, Burkholderia pseudomallei 1710b, Burkholderia pseudomallei 305, Burkholderia pseudomallei 406e, Burkholderia pseudomallei 668, Burkholderia pseudomallei 7894, Burkholderia pseudomallei K96243, Burkholderia sp. 383, Burkholderia thailandensis Bt4, Burkholderia thailandensis E264, Burkholderia thailandensis MSMB43, Burkholderia thailandensis TXDOH, Burkholderia ubonensis Bu, Burkholderia vietnamiensis G4, Caulobacter crescentus CB15, Cellvibrio japonicus Uedacterium 12, Chibabacterium 12, Chibium , Comamonas testosteroni KF-1, Cupriavidus taiwanensis, Dechloromonas aromatica RCB, Delftia acidovorans SPH-1, Desulfovibrio desulfuricans subsp. desulfuricans strain G20, Desulfovibrio desulfuricans subsp. desulfuricans strain G20, Enterobacter cancerogenus ATCC 35316, Enterobacter sakazakii ATCC BAA-894, Enterobacter sp. 638, Erwinia tasmaniensis, Escherichia albertii TW07627, Escherichia coli O157.H7 EDL933, Escherichia coli O157: H7 strain EC4024, Escherichia coli O157: H7 strain EC4196, gamma proteobacterium HTCC5015, gamma proteobacterium KT 71, sp. M21, Hahella chejuensis KCTC 2396, Halorhodospira halophila SL1, Herminiimonas arsenicoxydans, Bald idiomarina OS145, Idiomarina loihiensis L2TR, Janthinobacterium sp. Marseille, Klebsiella pneumoniae 342, Klebsiella pneumoniae subsp. pneumoniae MGH 78578, Klebsiella sp. ZD414, Leptospira interrogans serovar Copenhageni strain Fiocruz L1-130, Leptospira interrogans serovar Copenhageni strain 56601, Leptothrix cholodnii SP-6, Limnobacter sp. MED105, Magnetospirillum magneticum AMB-1, gamma proteobacterium marine HTCC2080, gamma proteobacterium marine HTCC2143, gamma proteobacterium marine HTCC2207, marine metagenome, Marinobacter algicola DG893, Marinobacter aquaeolei VT8, Marinobacter sp. ELB17, Marinomonas sp. MWYL1, Methylibium petroleiphilum PM1, Methylobacillus flagellatus KT, Methylobacterium chloromethanicum CM4, Methylobacterium extorquens PA1, Methylobacterium populi BJ001, Methylocella silvestris BL2, Methylococcus capsulatit strain Morpa Bath. PE36, Nitrobacter sp. Nb-311A, Nitrobacter winogradskyi Nb-255, Nitrococcus mobilis nb-231, Nitrosococcusoceani ATCC 19707, Nitrosococcus oceani C-27, Nitrosomonas europaea ATCC 19718, Nitrosomonas eutropha C91, Nitrosopira multiformis ATCC 25196 sp. Oceanospirill MED92, Oligotropha carboxidovorans OM5, Pectobacterium atrosepticum SCRI1043, Photobacterium profundum 3TCK, Photobacterium profundum SS9, Photobacterium sp. SKA34, Photorhabdus luminescens, Photorhabdus luminescens subsp. laumondii TTO1, Polaromonas naftalenivoranos CJ2, Polaromonas sp. JS666, Polynucleobacter sp. QLW-P1DMWA-1, Proteus mirabilis HI4320, Providencia stuartii ATCC 25827, Pseudoalteromonas atlantica T6c, Pseudoalteromonas haloplanktis TAC125, Pseudoalteromonas sp. 643A, Pseudoalteromonas tunicata D2, Pseudomonas aeruginosa PA7, Pseudomonas aeruginosa PACS2, Pseudomonas aeruginosa PAO1, Pseudomonas aeruginosa UCBPP-PA14, Pseudomonas entomophila L48, Pseudomonas fluorescens Pfududse Pseudonas Pf0-1, Pseudonas Pf0-1 GB-1, Pseudomonas putida KT2440, Pseudomonas putida W619, Pseudomonas stutzeri A1501, Pseudomonas syringae pv. Phaseolicola 1448A, Pseudomonas syringae pv. syringae B728a, Pseudomonas syringae pv. Tomato strain DC3000, Psychromonas ingrahamii 37, Ralstonia eutropha H16, Ralstonia eutropha JMP134, Ralstonia metallidurans CH34, Ralstonia pickettii 12D, Ralstonia pickettii 12J, Ralstonia solanacearum GMI1000, Ralstonia solanacearum IPO16cearumumumumum, Ralstonia solanaumumum, Ralstonia solanaumine Med297, Bacteria Rhodobacterales Y4I, Rhodoferax ferrireducens T118, Rhodopseudomona spalustris BisA53, Rhodopseudomonas palustris BisB18, Rhodopseudomonas palustris BisB5, Rhodopseudomonas palustris CGA009, Rhodopseudomonas palustris, Paistromonas paistris, Paistris-3 arizonae serovar 62: z4, z23: -, Salmonella enterica subsp. enterica serovar Choleraesuis strain SC-B67, Salmonella enterica subsp. Enterica serovar allinarum strain 287/91, Salmonella enterica subsp. Enterica serovar Hadar strain RI_05P066, Salmonella enterica subsp. enterica serovar Javiana strain GA_MM04042433, Salmonella enterica subsp. enterica serovar Saintpaul strain SARA23, Salmonella enterica subsp. enterica serovar Saintpaul strain SARA29, Salmonella enterica subsp. Enterica serovar Typhi strain CT18, Salmonella typhimurium LT2, Serratia proteamaculans 568, Shewanella amazonensis SB2B, Shewanella baltica OS155, Shewanella baltica OS185, Shewanella baltica OS195, Shewanella baltica OS223, Shewanella benthica KT99, Shewanella denitrificans OS217, Shewanella frigidimarina NCIMB 400, Shewanella halifaxensis HAW -EB4, Shewanella loihica PV-4, Shewanella oneidensis MR-1, Shewanella pealeana ATCC 700345, Shewanella putrefaciens 200, Shewanella sediminis HAW-EB3, Shewanella sp. ANA-3, Shewanella sp. MR-4, Shewanella sp. MR-7, Shewanella sp. W3-18-1, Shewanella woodyi ATCC 51908, Shigella boydii Sb227, Shigella dysenteriae Sd197, Stenotrophomonas maltophilia K279a, Stenotrophomonas maltophilia R551-3, Streptococcus sp. ( N1), synthetic construction, Thauera sp. MZ1T, Thioalkalivibrio sp. HL-EbGR7, Thiobacillus denitrificans ATCC25259, Thiomicrospira crunogena x CL-2, Vibrio alginolyticus 12G01, Vibrio angustum S14, Vibrio campbellii AND4, Vibrio cholerae 2740-80, Vibrio cholerae MZO-2, Vibrio cholerae O1 biovar61 cholera61, cholera61 Ov biolera61, cholera61, N9 biovator, N9 biovator, Nr. , Vibrio fischeri ES114, Vibrio fischeri MJ11, Vibrio harveyi ATCC BAA-1116, Vibrio mimicus, Vibrionales bacterium SWAT-3, Vibrio parahaemolyticus AQ3810, Vibrio parahaemolyticus RIMD 2210633, Vibrio shilonii AK1, Vibrio splendidus 12B01, Vibrio splendidus 12. MED222, Vibrio vulnificus CMCP6, Vibrio vulnificus YJ016, Xanthomonas axonopodis pv. citri strain 306, Xanthomonas campestris pv. campestris strain ATCC 33913, Xanthomonas campestris pv. campestris strain B100, Xanthomonas campestris pv. Vesicatoria strain 85-10, Xanthomonas oryzae pv. oryzae KACC10331, Xanthomonas oryzae pv. oryzae PXO99A, Xanthomonas oryzae pv. oryzicola BLS256, Yersinia bercovieri ATCC 43970, Yersinia enterocolitica subsp. enterocolitica 8081, Yersinia frederiksenii ATCC 33641, Yersinia intermedia ATCC 29909, Yersinia mollaretii ATCC 43969, Yersinia pestis Angola, Yersinia pestis biovar Orientalis strain F1991016, Yersinia pestis CO92, Yersinia pestis KIM or Yersinia pseudotuberculosis 31.

En un aspecto, la tioesterasa precursora es una tioesterasa que tiene una secuencia analoga a la de 'TesA (por ejemplo, una enzima TesA sin el peptido senal). La tioesterasa precursora puede tener al menos aproximadamente 20 %, por ejemplo, al menos aproximadamente 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 96 %, 97 %, 98 % o 99 % de identidad de secuencia con 'TesA. En otro ejemplo mas, la tioesterasa precursora tiene al menos aproximadamente 20 %, por ejemplo, al menos aproximadamente 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 96 %, 97 %, 98% o 99% de identidad de secuencia con una 'TesA que se obtiene de una bacteria E. coli, tal como, una bacteria E.coli K12. En un ejemplo adicional, la tioesterasa precursora es una tioesterasa que tiene una secuencia analoga a la secuencia de la SEC ID NO: 31 expuesta en la FIG. 58, y preferentemente al menos aproximadamente 20 %, por ejemplo, al menos aproximadamente 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 96 %, 97 %, 98% o 99% de identidad de secuencia con la SEQ ID NO: 31 expuesta en la FIG. 58. La secuencia analoga puede ser de una proteina de origen natural o puede ser de una proteina modificada previamente.

In one aspect, the precursor thioesterase is a thioesterase having a sequence analogous to that of 'TesA (for example, a TesA enzyme without the signal peptide). The precursor thioesterase may have at least about 20%, for example, at least about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with 'TesA. In another example, the precursor thioesterase has at least about 20%, for example, at least about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70% , 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with a 'TesA that is obtained of an E. coli bacteria , such as an E.coli K12 bacteria . In a further example, the precursor thioesterase is a thioesterase having a sequence analogous to the sequence of SEQ ID NO: 31 set forth in FIG. 58, and preferably at least about 20%, for example, at least about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80 %, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 31 set forth in FIG. 58. The analogous sequence can be from a naturally occurring protein or it can be from a previously modified protein.

En un aspecto, la tioesterasa precursora es una tioesterasa que comprende las siguientes series de aminoacidos:

In one aspect, the precursor thioesterase is a thioesterase comprising the following series of amino acids:

G-D-S-L-X(5)-M (SEQ ID NO:28), en la que:

G-D-S-L-X (5) -M (SEQ ID NO: 28), in which:

la "X" se refiere a cualquier resto de aminoacido; el numero en el parentesis adyacente al mismo, cuando esta presente, se refiere al numero de restos X en el tramo de restos de aminoacidos;

"X" refers to any amino acid residue; the number in the parentheses adjacent to it, when present, refers to the number of residues X in the amino acid residue segment;

el resto S en la posicion 3 es un resto catalitico;

the remainder S in position 3 is a catalytic remainder;

el resto D en la posicion 2 puede estar sustituido con N o T;

the remainder D in position 2 may be substituted with N or T;

el resto L en la posicion 4 puede estar sustituido con C o Q;

the remainder L in position 4 may be substituted with C or Q;

el resto M en la posicion 10 puede estar sustituido con C, D, L, N, T o V;

the remainder M in position 10 may be substituted with C, D, L, N, T or V;

V-X(2)-G-X-N-D-X-L (SEQ ID NO:29), en la que:

V-X (2) -G-X-N-D-X-L (SEQ ID NO: 29), in which:

cada "X" se refiere a cualquier resto de aminoacido; el numero en el parentesis adyacente al mismo, cuando esta presente, se refiere al numero de restos X en el tramo de restos de aminoacidos;

each "X" refers to any amino acid residue; the number in the parentheses adjacent to it, when present, refers to the number of residues X in the amino acid residue segment;

el resto N en la posicion 6 esta en el orificio oxianionico;

the remainder N in position 6 is in the oxyanionic hole;

el resto V en la posicion 1 puede estar sustituido con L;

the remainder V in position 1 may be substituted with L;

el resto N en la posicion 6 puede estar sustituido con V, L, C, A, G, H, I, T o W;

the remainder N in position 6 may be substituted with V, L, C, A, G, H, I, T or W;

el resto D en la posicion 7 puede estar sustituido con E;

the remainder D in position 7 may be substituted with E;

el resto L en la posicion 9 puede estar sustituido con I, W, F, T, M, A, E, N o V;

the remainder L in position 9 may be substituted with I, W, F, T, M, A, E, N or V;

D-X(2)-H-P-X(7)-I (SEQ ID NO:30), en la que:

D-X (2) -H-P-X (7) -I (SEQ ID NO: 30), in which:

cada "X" se refiere a cualquier resto de aminoacido; cada numero en el parentesis adyacente al mismo, cuando esta presente,

each "X" refers to any amino acid residue; each number in the parentheses adjacent to it, when present,

se refiere al numero de restos X en el tramo respectivo de restos de aminoacidos;

refers to the number of residues X in the respective stretch of amino acid residues;

los restos D y H en las posiciones 1 y 4 respectivamente son los restos cataliticos;

residues D and H in positions 1 and 4 respectively are the catalytic residues;

el resto P en la posicion 5 puede estar sustituido con G, A, F, L, S o V;

the remainder P in position 5 may be substituted with G, A, F, L, S or V;

el resto I en la posicion 13 puede estar sustituido con L o V.

the remainder I in position 13 may be substituted with L or V.

En un aspecto, la tioesterasa precursora es una tioesterasa que tiene inmunorreactividad cruzada con una 'TesA obtenida de una bacteria E. coli. En un aspecto particular, la tioesterasa precursora tiene inmunorreactividad cruzada con la 'TesA obtenida de una bacteria E. coli K-12. En un aspecto particular, la tioesterasa precursora tiene inmunorreactividad cruzada con una tioesterasa que comprende la secuencia de aminoacidos de la SEC ID NO: 31 como se expone en la FIG. 58. En un aspecto particular, la tioesterasa precursora tiene reactividad cruzada con fragmentos (o partes) de cualquiera de las tioesterasas obtenidas de una bacteria E. coli, o de una bacteria E. coli K-12 y/o de cualquier tioesterasa que comprenda la secuencia de aminoacidos de la SEC ID NO: 31 como se expone en la FIG. 58. La enzima precursora que tiene inmunorreactividad cruzada con 'TesA puede ser una proteina de origen natural, una proteina previamente modificada, o una proteina sintetica.

In one aspect, the precursor thioesterase is a thioesterase that has cross-immunoreactivity with a 'TesA obtained from an E. coli bacteria . In a particular aspect, the precursor thioesterase has cross-immunoreactivity with the 'TesA obtained from an E. coli K-12 bacteria. In a particular aspect, the precursor thioesterase has cross-immunoreactivity with a thioesterase comprising the amino acid sequence of SEQ ID NO: 31 as set forth in FIG. 58. In a particular aspect, the precursor thioesterase has cross-reactivity with fragments (or parts) of any of the thioesrases obtained from an E. coli bacterium , or from an E. coli K-12 bacterium and / or from any thioesterase comprising the amino acid sequence of SEQ ID NO: 31 as set forth in FIG. 58. The precursor enzyme that has cross-immunoreactivity with 'TesA may be a naturally occurring protein, a previously modified protein, or a synthetic protein.

En otro ejemplo particular, la tioesterasa precursora es una 'TesA de una bacteria E. coli, o es un homologo, un paralogo o un ortologo de una 'TesA de una bacteria E. coli, tal como una 'TesA de una bacteria E. coli K12. La tioesterasa precursora de la que procede un mutante de la presente divulgacion, tambien puede ser una parte o un fragmento enzimaticamente activa(o) de una cualquiera de las tioesterasas descritas anteriormente.

In another particular example, the precursor thioesterase is a 'TesA of an E. coli bacteria , or is a homologue, a paralogue or an ortholog of a' TesA of an E. coli bacteria , such as a 'TesA of an E. bacteria . coli K12. The precursor thioesterase from which a mutant of the present disclosure is derived may also be an enzymatically active part or fragment of any one of the thioesrases described above.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales) que comprende una secuencia de aminoacidos que tiene al menos una sustitucion de un aminoacido, en comparacion con una tioesterasa precursora, de modo que la tioesterasa mutante tiene al menos una propiedad alterada en relacion con la tioesterasa precursora. En un aspecto ejemplar, se desvela una tioesterasa mutante que tiene una secuencia de aminoacidos con una sola mutacion por sustitucion y muestra al menos una propiedad alterada en comparacion con la tioesterasa precursora de la que procede la mutante. En un aspecto ejemplar, se desvela una tioesterasa mutante que comprende una secuencia de aminoacidos que tiene dos o mas mutaciones por sustitucion a partir de la secuencia de su tioesterasa precursora, y la tioesterasa mutante tiene al menos una propiedad alterada en comparacion con la tioesterasa precursora.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed which comprises an amino acid sequence that has at least one amino acid substitution, as compared to a precursor thioesterase, so that the mutant thioesterase has at least one altered property in relation to the precursor thioesterase. In an exemplary aspect, a mutant thioesterase is disclosed that has an amino acid sequence with a single substitution mutation and shows at least one altered property compared to the precursor thioesterase from which the mutant is derived. In an exemplary aspect, a mutant thioesterase is disclosed comprising an amino acid sequence having two or more substitution mutations from the sequence of its precursor thioesterase, and the mutant thioesterase has at least one altered property compared to the precursor thioesterase. .

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que es una variante de una tioesterasa precursora, y que tiene al menos una propiedad alterada in vitro o en vivo en relacion con dichas tioesterasa precursora, en donde la tioesterasa precursora es una tioesterasa que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58 y, por consiguiente, comprende los restos de aminoacidos correspondientes 1-182 de la s Ec ID NO: 31, y en donde la tioesterasa precursora se modifica en una o mas posiciones de aminoacidos seleccionadas de las posiciones correspondientes a uno o mas restos 1-182 de la SEC ID NO: 31 expuesta en la FIG. 58.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which is a variant of a precursor thioesterase, and which has at least one property altered in vitro or in vivo in relation to said precursor thioesterase, wherein the precursor thioesterase is a thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58 and, consequently, comprises the corresponding amino acid residues 1-182 of s Ec ID NO: 31, and wherein the precursor thioesterase is modified at one or more amino acid positions selected from the positions corresponding to one or more residues 1 -182 of SEQ ID NO: 31 set forth in FIG. 58.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58 y, por consiguiente, comprende los correspondientes restos de aminoacidos 1-182 de la SEC ID NO: 31, y que tiene al menos una propiedad alterada in vitro o in vivo en relacion con dicha tioesterasa precursora, en donde la tioesterasa precursora se muta en una o mas posiciones, correspondientes a una o mas posiciones de aminoacidos de la SEC ID NO: 31 expuesta en la FIG. 58, seleccionadas de las posiciones 1, 2, 3, 4, 5, 6 , 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181 y/o 182.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58 and, therefore, comprises the corresponding amino acid residues 1-182 of SEQ ID NO: 31, and having at least one property altered in vitro or in vivo in relation to said precursor thioesterase, wherein the precursor thioesterase is mutated in one or more positions, corresponding to one or more amino acid positions of SEQ ID NO: 31 set forth in FIG. 58, selected from positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41.42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181 and / or 182.

En un aspecto, se desvela una tioesterasa mutante (o un equivalente natural), que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58 y, por consiguiente, comprende los correspondientes restos de aminoacidos 1-182 de la SEC ID NO: 31, y que tiene al menos una propiedad alterada in vitro o in vivo en relacion con dicha tioesterasa precursora, en donde la tioesterasa precursora se muta con una o mas sustituciones seleccionadas de A1C, A1F, A1L, A1Q, A1R, A1S, A1V, A1Y, D2E, D2H, D2K, D2L, D2M, D2P, D2R, D2W, T3E, T3G, T3K, T3L, T3R, T3W, L4A, L4G, L4M, L4N, L4S, L4V, L4Y, L5C, L5E, L5F, L5G, L5H, L5K, L5N, L5Q, L5S, L5W, L5Y, I6A, I6L, I6T, I6V, L7A, L7C, L7E, L7K, L7M, L7N, L7S, L7T, L7V, L7W, L7Y, G8A, G8K, G8S, D9N, D9T, S10C, L11A, L11C, L11I, L11M, L11Q, L11V, S12A, S12I, S12L, S12M, S12N, S12T, S12V, S12Y, A13C, A13D, A13G, A13H, A13I, A13L, A13N, A13S, A13T, A13V, A13W, A13Y, G14A, G14C, G14E, G14F, G14I, G14K, G14M, G14N, G14P, G14Q, G14R, G14S, G14T, G14V, Y15A, Y15C, Y15D, Y15E, Y15G, Y15I, Y15L, Y15M, Y15N, Y15Q, Y15R, Y15S, Y15V, R16A, R16D, R16E, R16G, R16H, R16I, R16L, R16M, R16N, R16P, R16Q, R16S, R16T, R16V, R16W, M17A, M17C, M17D, M17E, M17G, M17K, M17L, M17N, M17P, M17Q, M17R, M17S, M17T, M17V, S18E, S18M, S18N, S18T, A19C, A19E, A19L, A19V, S20A, S20C, S20D, S20G, S20L, S20T, S20W, A21G, A21I, A21L, A21P, A21Y, A22C, A22D, A22E, A22F, A22G, A22H, A22I, A22K, A22L, A22M, A22N, A22P, A22R, A22S, A22T, A22Y, W23A, W23H, W23N, W23P, W23Y, P24A, P24C, P24D, P24E, P24F, P24G, P24I, P24M, P24N, P24S, P24T, P24V, P24W, A25D, A25E, A25L, A25N, A25P, A25Q, A25R, A25S, A25V, L26C, L26D, L26E, L26F, L26G, L26H, L26I, L26K, L26N, L26P, L26Q, L26R, L26S, L26V, L26W, L26Y, L27A, L27C, L27F, L27H, L27M, L27R, L27S, L27T, L27V, L27W, L27Y, N28A, N28G, N28I, N28K, N28M, N28P, N28R, N28W, D29M, D29P, D29V, K30P, W31D, W31E, W31G, W31L, W31N, W31P, W31R, W31S, W31T, Q32V, Q32Y, S33F, S33G, S33I, S33M, S33R, K34A, K34H, K34M, K34R, T35F, T35G, T35K, T35L, T35M, T35Q, T35V, T35Y, S36A, S36F, S36H, S36I, S36L, S36W, V37A, V37F, V37G, V37H, V37L, V37N, V37S, V37Q, V37S, V37W, V37Y, V38D, V38E, V38F, V38G, V38K, V38L, V38P, V38R, V38S, N39A, N39C, N39E, N39F, N39G, N39K, N39M, N39P, N39Q, N39R, N39T, N39V, N39W, N39Y, A40D, A40G, A40H, A40L, A40M, A40P, A40T, A40V, A40Y, S41C, S41P, S41T, I42A, I42C, I42D, I42E, I42G, I42K, I42L, I42M, I42P, I42S, I42T, I42W, I42Y, S43A, S43C, S43D, S43E, S43F, S43G, S43H, S43L, S43M, S43N, S43P, S43R, S43T, S43V, S43W, G44A, G44C, G44E, G44F, G44H, G44K, G44L, G44M, G44N, G44Q, G44R, G44S, G44W, G44Y, D45A, D45C, D45E, D45F, D45G, D45H, D45I, D45K, D45L, D45M, D45P, D45Q, D45S, D45T, D45V, D45W, T46A, T46C, T46D, T46E, T46F, T46G, T46I, T46K, T46L, T46N, T46R, T46S, T46V, T46W, S47A, S47C, S47E, S47F, S47G, S47L, S47M, S47P, S47Q, S47R, S47T, S47V, S47W, S47Y, Q48C, Q48D, Q48E, Q48F, Q48G, Q48I, Q48M, Q48S, Q48T, Q48V, Q48W, Q48Y, Q49A, Q49C, Q49D, Q49E, Q49G, Q49H, Q49I, Q49K, Q49L, Q49M, Q49P, Q49R, Q49S, Q49V, Q49W, Q49Y, G50A, G50C, G50E, G50F, G50I, G50K, G50L, G50M, G50N, G50P, G50Q, G50R, G50S, G50T, G50W, G50Y, L51A, L51C, L51D, L51F, L51H, L51N, L51P, L51S, L51T, L51V, L51W, L51Y, A52C, A52D, A52H, A52I, A52L, A52M, A52P, A52R, A52V, A52W, A52Y, R53A, R53C, R53D, R53E, R53F, R53G, R53I, R53K, R53L, R53N, R53S, R53T, R53V, R53W, R53Y, L54A, L54C, L54E, L54F, L54G, L54M, L54N, L54S, L54T, L54W, L54Y, P55A, P55G, P55Y, A56P, A56R, A56W, A56Y, L57A, L57C, L57F, L57G, L57H, L57I, L57K, L57N, L57P, L57Q, L57R, L57S, L57T, L57V, L57W, L57Y, L58A, L58D, L58E, L58F, L58G, L58H, L58I, L58M, L58N, L58R, L58S, L58V, L58W, L58Y, K59E, K59R, K59V, Q60E, Q60M, Q60P, H61A, H61D, H61E, H61G, H61P, H61W, Q62G, Q62M, Q62P, Q62W, P63D, P63E, P63G, P63I, P63K, P63L, P63M, P63N, P63Q, P63R, P63S, P63T, P63V, P63W, R64D, R64E, R64F, R64L, R64M, R64P, R64Q, R64W, R64Y, W65A, W65E, W65G, W65K, W65L, W65M, W65N, W65P, W65R, W65V, V66C, V66G, V66I, V66M, V66N, V66Q, V66S, V66W, V66Y, L67A, L67C, L67E, L67G, L67M, L67Q, L67Q, L67S, L67T, L67W, V68A, V68E, V68G, V68L, V68M, V68N, V68P, V68Q, V68S, V68T, E69A, E69C, E69D, E69F, E69G, E69H, E69K, E69L, E69M, E69N, E69P, E69Q, E69S, E69V, E69W, E69Y, L70A, L70C, L70E, L70F, L70G, L70H, L70I, L70K, L70Q, L70S, L70T, L70V, L70W, G71A, G71C, G71S, G72A, G72C, G72M, G72P, G72S, N73A, N73C, N73G, N73H, N73I, N73L, N73P, N73R, N73S, N73T, N73V, N73W, D74A, D74C, D74E, D74F, D74G, D74Q, D74S, D74W, D74Y, G75A, G75C, G75D, G75E, G75F, G75I, G75K, G75L, G75M, G75N, G75P, G75R, G75T, G75V, G75W, G75Y, L76A, L76C, L76D, L76E, L76F, L76G, L76I, L76K, L76M, L76N, L76P, L76Q, L76R, L76T, L76V, L76W, R77A, R77C, R77D, R77E, R77F, R77G, R77H, R77K, R77L, R77N, R77Q, R77S, R77V, R77W, G78A, G78C, G78D, G78E, G78F, G78M, G78N, G78P, G78Q, G78R, G78S, G78T, G78V, G78Y, F79A, F79D, F79E, F79G, F79H, F79K, F79M, F79N, F79P, F79Q, F79S, F79V, F79W, F79Y, Q80A, Q80E, Q80G, Q80L, Q80M, Q80S, Q80W, Q80Y, P81A, P81E, P81K, P81L, P81M, P81N, P81T, P81W, P81Y, Q82A, Q82F, Q82I, Q82M, Q82N, Q82P, Q82R, Q82S, Q82T, Q82V, Q82W, Q82Y, Q83A, Q83C, Q83F, Q83G, Q83K, Q83L, Q83M, Q83N, Q83R, Q83S, Q83T, Q83V, Q83W, Q83Y, T84A, T84D, T84E, T84F, T84G, T84H, T84K, T84L, T84M, T84N, T84Q, T84R, T84S, T84V, T84W, T84Y, E85A, E85C, E85D, E85F, E85G, E85L, E85P, E85Q, E85R, E85S, E85T, E85V, E85W, E85Y, Q86A, Q86G, Q86H, Q86K, Q86P, Q86T, Q86V, Q86W, Q86Y, T87A, T87C, T87D, T87E, T87F, T87G, T87H, T87L, T87M, T87P, T87R, T87S, T87V, T87W, L88A, L88C, L88E, L88F, L88G, L88H, L88Q, L88S, L88W, L88Y, R89A, R89G, R89H, R89L, R89P, R89T, R89V, R89W, Q90E, Q90L, Q90N, Q90P, Q90W, Q90Y, I91E, I91G, I91L, I91M, I91N, I91Q, I91S, I91V, I91Y, L92A, L92C, L92E, L92G, L92H, L92N, L92Q, L92R, L92S, L92T, L92V, L92Y, Q93A, Q93E, Q93F, Q93G, Q93H, Q93I, Q93L, Q93M, Q93N, Q93P, Q93S, Q93V, Q93W, Q93Y, D94C, D94E, D94F, D94G, D94H, D94K, D94L, D94N, D94P, D94Q, D94R, D94S, D94V, V95A, V95C, V95D, V95E, V95F, V95G, V95I, V95L, V95M, V95N, V95P, V95Q, V95T, V95W, V95Y, K96A, K96C, K96L, K96N, K96P, K96Q, K96R, K96V, K96Y, A97C, A97E, A97F, A97K, A97N, A97P, A97R, A97V, A97W, A98E, A98G, A98K, A98L, A98P, A98V, A98W, A98Y, N99A, N99C, N99D, N99G, N99L, N99M, N99P, N99Q, N99R, N99S, N99W, N99Y, A100D, A100E, A100G, A100H, A100I, A100K, A100L, A100M, A100Q, A100R, A100S, A100T, A100V, A100W, A100Y, E101A, E101D, E101G, E101L, E101M, E101P, E101S, E101T, E101V, P102E, P102F, P102G, P102H, P102I, P102L, P102Q, P102R, P102S, P102V, P102W, P102Y, L103A, L103C, L103E, L103G, L103I, L103K, L103N, L103Q, L103R, L103S, L103T, L103V, L103W, L104A, L104C, L104E, L104G, L104I, L104N, L104P, L104Q, L104S, L104W, L104Y, M105A, M105C, M105E, M105F, M105G, M105I, M105K, M105L, M105P, M105T, M105V, M105W, Q106A, Q106C, Q106D, Q106G, Q106H, Q106K, Q106L, Q106M, Q106R, Q106S, Q106T, Q106V, Q106W, Q106Y, I107A, I107C, I107E, I107F, I107G, I107K, I107L, I107M, I107Q, I107S, I107T, I107V, I107Y, R108A, R108C, R108D, R108E, R108F, R108G, R108H, R108I, R108L, R108M, R108S, R108V, R108W, R108Y, L109A, L109C, L109D, L109E, L109F, L109G, L109K, L109M, L109P, L109Q, L109R, L109S, L109T, L109V, L109Y, P110A, P110C, P110D, P110E, P110F, P110G, P110H, P110K, P110L, P110M, P110N, P110R, P110S, P110V, P110W, A111C, A111E, A111L, A111M, A111N, A111P, A111Q, A111R, A111S, A111V, A111W, A111Y, N112A, N112F, N112G, N112I, N112K, N112L, N112P, N112R, N112V, N112W, N112Y, Y113A, Y113C, Y113D, Y113E, Y113G, Y113I, Y113M, Y113P, Y113Q, Y113S, Y113S, Y113W, G114A, G114F, G114K, G114L, G114M, G114P, G114W, G114Y, R115A, R115C, R115E, R115G, R115I, R115N, R115P, R115Q, R115S, R115V, R115W, R115Y, R116C, R116D, R116E, R116H, R116T, R116V, R116W, Y117A, Y117C, Y117D, Y117E, Y117G, Y117H, Y117I, Y117L, Y117M, Y117N, Y117P, Y117Q, Y117R, Y117S, Y117T, Y117V, Y117W, N118A, N118C, N118E, N118F, N118G, N118H, N118I, N118K, N118L, N118M, N118P, N118Q, N118S, N118T, N118V, N118W, E119C, E119D, E119F, E119G, E119K, E119L, E119M, E119P, E119Q, E119R, E119T, E119W, E119Y, A120D, A120E, A120G, A120I, A120L, A120P, A120T, A120W, F121A, F121C, F121D, F121E, F121G, F121K, F121L, F121M, F121N, F121P, F121Q, F121R, F121S, F121V, F121W, F121Y, S122A, S122C, S122D, S122E, S122F, S122G, S122I, S122L, S122M, S122P, S122R, S122V, S122W, S122Y, A123C, A123E, A123F, A123H, A123L, A123R, A123T, A123V, A123W, A123Y, I124A, I124C, I124D, I124E, I124G, I124H, I124K, I124L, I124R, I124S, I124T, I124W, I124Y, Y125C, Y125F, Y125G, Y125H, Y125I, Y125L, Y125P, Y125Q, Y125R, Y125S, Y125T, Y125V, Y125W, P126C, P126F, P126H, P126K, P126R, P126T, P126V, P126Y, K127A, K127I, K127P, K127S, L128A, L128C, L128E, L128F, L128G, L128Q, L128R, L128S, L128T, L128V, L128W, A129D, A129F, A129H, A129I, A129K, A129L, A129N, A129W, A129Y, K130E, K130I, K130P, K130V, E131A, E131C, E131D, E131F, E131G, E131I, E131K, E131L, E131N, E131P, E131V, E131W, F132C, F132D, F132E, F132K, F132L, F132N, F132P, F132T, F132V, D133C, D133K, D133R, D133S, D133T, D133V, D133Y, V134C, V134D, V134E, V134I, V134K, V134M, V134N, V134P, V134Q, V134R, V134S, V134W, V134Y, P135A, P135E, P135K, P135Q, L136A, L136C, L136D, L136E, L136F, L136G, L136H, L136K, L136M, L136N, L136P, L136Q, L136R, L136S, L136T, L137A, L137C, L137D, L137E, L137G, L137H, L137K, L137P, L137Q, L137R, L137S, L137Y, P138E, P138F, P138G, P138N, P138R, P138T, P138V, F139A, F139C, F139D, F139E, F139G, F139H, F139L, F139M, F139N, F139S, F139T, F139V, F139W, F140A, F140C, F140G, F140I, F140L, F140M, F140N, F140P, F140S, F140T, F140V, F140W, M141A, M141C, M141D, M141E, M141F, M141G, M141K, M141L, M141P, M141Q, M141R, M141T, M141V, M141W, M141Y, E142A, E142C, E142G, E142I, E142L, E142M, E142N, E142P, E142Q, E142R, E142S, E142T, E142V, E142W, E142Y, E143A, E143D, E143F, E143G, E143I, E143M, E143P, E143W, V144A, V144D, V144E, V144G, V144H, V144N, V144P, V144Q, V144R, V144S, V144W, V144Y, Y145A, Y145C, Y145D, Y145E, Y145G, Y145I, Y145L, Y145M, Y145N, Y145Q, Y145R, Y145S, Y145T, Y145W, L146A, L146C, L146D, L146E, L146G, L146H, L146P, L146S, L146W, K147G, K147P, K147R, K147W, P148D, P148E, P148W, Q149L, W150C, W150D, W150E, W150G, W150L, W150P, W150Q, W150R, W150T, M150V, M151A, M151C, M151D, M151E, M151F, M151G, M151I, M151L, M151Q, M151R, M151S, M151T, M151V, M151W, Q152A, Q152D, Q152E, Q152F, Q152H, Q152I, Q152K, Q152L, Q152N, Q152P, Q152R, Q152S, Q152T, Q152V, Q152Y, D153A, D153E, D153F, D153I, D153K, D153M, D153P, D153Q, D153V, D153W, D154A, D154C, D154E, D154F, D154G, D154H, D154I, D154K, D154L, D154M, D154N, D154P, D154R, D154S, D154T, D154V, D154W, G155A, G155F, G155H, G155I, G155P, G155V, G155W, G155Y, I156A, I156C, I156E, I156F, I156G, I156K, I156L, I156M, I156Q, I156R, I156S, I156T, I156V, I156Y, H157C, H157E, P158A, P158F, P158G, P158H, P158I, P158L, P158Q, P158S, P158T, P158V, P158W, N159C, N159E, N159G, N159I, N159K, N159L, N159M, N159P, N159Q, N159R, N159T, N159V, N159W, R160A, R160C, R160D, R160E, R160G, R160H, R160I, R160K, R160N, R160Q, R160S, R160W, D161E, D161G, D161I, D161K, D161L, D161M, D161N, D161Q, D161R, D161S, D161V, D161W, A162G, A162I, A162K, A162L, A162N, A162R, A162T, A162V, A162Y, Q163A, Q163C, Q163D, Q163E, Q163F, Q163G, Q163I, Q163L, Q163M, Q163S, Q163T, Q163V, Q163W, Q163Y, P164A, P164C, P164D, P164K, P164L, P164M, P164N, P164R, P164T, P164V, P164W, F165D, F165E, F165G, F165H, F165I, F165K, F165L, F165M, F165R, F165S, F165T, F165V, F165W, F165Y, I166A, I166C, I166F, I166L, I166M, I166S, I166V, I166Y, A167C, A167D, A167E, A167F, A167G, A167K, A167L, A167M, A167N, A167Q, A167R, A167T, A167V, A167W, A167Y, D168A, D168G, D168H, D168L, D168M, D168P, D168R, D168T, D168V, D168W, W169A, W169D, W169E, W169G, W169K, W169M, W169Q, W169R, W169S, W169T, W169V, M170A, M170E, M170F, M170G, M170H, M170L, M170N, M170Q, M170S, M170T, M170V, M170W, M170Y, A171E, A171F, A171I, A171S, A171V, A171W, K172A, K172M, K172P, Q173D, Q173I, Q173N, Q173P, Q173W, Q173Y, L174A, L174F, L174G, L174Q, L174S, L174T, L174W, L174W, L174Y, Q175F, Q175I, Q175L, Q175M, Q175Y, P176D, P176H, P176K, P176L, P176N, P176Q, P176R, P176V, P176W, P176Y, L177D, L177F, L177G, L177M, L177S, L177T, V178A, V178F, V178G, V178K, V178L, V178R, V178S, V178T, V178W, N179G, N179H, N179R, N179T, N179V, N179W, N179Y, H180A, H180E, H180G, H180L, H180P, H180R, H180S, H180V, H180W, D181A, D181C, D181E, D181G, D181H, D181I, D181L, D181P, D181Q, D181R, D181S, D181T, D181W, S182A, S182C, S182D, S182E, S182G, S182I, S182K, S182L, S182N, S182P, S182Q, S182R, S182T y/o S180V, en donde, en las designaciones de mutacion por sustitucion, los numeros se refieren a posiciones de aminoacidos de la SEC ID NO: 31.

In one aspect, a mutant thioesterase (or a natural equivalent) is disclosed, which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58 and, therefore, comprises the corresponding amino acid residues 1-182 of SEQ ID NO: 31, and having at least one property altered in vitro or in vivo in relation to said precursor thioesterase, wherein the precursor thioesterase is mutated with one or more substitutions selected from A1C, A1F, A1L, A1Q, A1R, A1S, A1V, A1Y, D2E, D2H, D2K, D2L, D2M, D2P, D2R, D2W, T3E, T3G, T3K, T3L, T3R, T3W , L4A, L4G, L4M, L4N, L4S, L4V, L4Y, L5C, L5E, L5F, L5G, L5H, L5K, L5N, L5Q, L5S, L5W, L5Y, I6A, I6L, I6T, I6V, L7A, L7C, L7C, L7C , L7K, L7M, L7N, L7S, L7T, L7V, L7W, L7Y, G8A, G8K, G8S, D9N, D9T, S10C, L11A, L11C, L11I, L11M, L11Q, L11V, S12A, S12I, S12L, S12M, S12M, S12M , S12T, S12V, S12Y, A13C, A13D, A13G, A13H, A13I, A13L, A13N, A13S, A13T, A13V, A13W, A13Y, G14A, G14C, G14E, G14F, G14I, G14K, G14M, G14N, G14P, G14P, , G14R, G14S, G14T, G14V, Y15A, Y15C, Y15D, Y15E, Y15G, Y15I, Y15L, Y15M, Y15N, Y15Q, Y15R, Y15S, Y15V, R16A, R16D, R16E, R16G, R16H, R16I, R16L, R16L , R16N, R16P, R16Q, R16S, R16T, R16V, R16W, M17A, M17C, M17D, M17E, M17G, M17K, M17L, M17N, M17P, M17Q, M17R, M17S, M17T, M17V, S18E, S18M, S18N, S18T, A19C, A19E, A19L, A19L, A19L, A19L, A19L, A19L, A19L, A19L S20A, S20C, S20D, S20G, S20L, S20T, S20W, A21G, A21I, A21L, A21P, A21Y, A22C, A22D, A22E, A22F, A22G, A22H, A22I, A22K, A22L, A22M, A22N, A22P, A22P, A22P, A22P, A22P A22S, A22T, A22Y, W23A, W23H, W23N, W23P, W23Y, P24A, P24C, P24D, P24E, P24F, P24G, P24I, P24M, P24N, P24S, P24T, P24V, P24W, A25D, A25E, A25L, A25L, A25L A25P, A25Q, A25R, A25S, A25V, L26C, L26D, L26E, L26F, L26G, L26H, L26I, L26K, L26N, L26P, L26Q, L26R, L26S, L26V, L26W, L26Y, L27A, L27C, L27F, L27F, L27F, L27F L27M, L27R, L27S, L27T, L27V, L27W, L27Y, N28A, N28G, N28I, N28K, N28M, N28P, N28R, N28W, D29M, D29P, D29V, K30P, W31D, W31E, W31G, W31L, W31N, W31N, W31N, W31N W31R, W31S, W31T, Q32V, Q32Y, S33F, S33G, S33I, S33M, S33R, K34A, K34H, K34M, K34R, T35F, T35G, T35K, T35L, T35M, T35Q, T35V, T35Y, S36A, S36F, S36F, S36F, S36F S36I, S36L, S36W, V37A, V37F, V37G, V37H, V37L, V37N, V37S, V37Q, V37S, V37W, V37Y, V38D, V38E, V38F , V38G, V38K, V38L, V38P, V38R, V38S, N39A, N39C, N39E, N39F, N39G, N39K, N39M, N39P, N39Q, N39R, N39T, N39V, N39W, N39Y, A40D, A40G, A40H, A40L, A40L, A40L , A40P, A40T, A40V, A40Y, S41C, S41P, S41T, I42A, I42C, I42D, I42E, I42G, I42K, I42L, I42M, I42P, I42S, I42T, I42W, I42Y, S43A, S43C, S43D, S43E, S43E, S43E, S43E , S43G, S43H, S43L, S43M, S43N, S43P, S43R, S43T, S43V, S43W, G44A, G44C, G44E, G44F, G44H, G44K, G44L, G44M, G44N, G44Q, G44R, G44S, G44W, G44Y , D45C, D45E, D45F, D45G, D45H, D45I, D45K, D45L, D45M, D45P, D45Q, D45S, D45T, D45V, D45W, T46A, T46C, T46D, T46E, T46F, T46G, T46I, T46K, T46L, T46L, T46L , T46R, T46S, T46V, T46W, S47A, S47C, S47E, S47F, S47G, S47L, S47M, S47P, S47Q, S47R, S47T, S47V, S47W, S47Y, Q48C, Q48D, Q48E, Q48F, Q48G, Q48I, Q48I, Q48I, Q48I , Q48S, Q48T, Q48V, Q48W, Q48Y, Q49A, Q49C, Q49D, Q49E, Q49G, Q49H, Q49I, Q49K, Q49L, Q49M, Q49P, Q49R, Q49S, Q49V, Q49W, Q49Y, G50A, G50C, G50E, G50E , G50I, G50K, G50L, G50M, G50N, G50P, G50Q, G50R, G50S, G50T, G50W, G50Y, L51A, L51C, L51D, L51F, L5 1H, L51N, L51P, L51S, L51T, L51V, L51W, L51Y, A52C, A52D, A52H, A52I, A52L, A52M, A52P, A52R, A52V, A52W, A52Y, R53A, R53C, R53D, R53E, R53F, R53G, R53F, R53G R53I, R53K, R53L, R53N, R53S, R53T, R53V, R53W, R53Y, L54A, L54C, L54E, L54F, L54G, L54M, L54N, L54S, L54T, L54W, L54Y, P55A, P55G, P55Y, A56P, A56R, A56W, A56Y, L57A, L57C, L57F, L57G, L57H, L57I, L57K, L57N, L57P, L57Q, L57R, L57S, L57T, L57V, L57W, L57Y, L58A, L58A, L58A, L58A, L58A, L58A, L58A, L58A L58E, L58F, L58G, L58H, L58I, L58M, L58N, L58R, L58S, L58V, L58W, L58Y, K59E, K59R, K59V, Q60E, Q60M, Q60P, H61A, H61D, H61E, H61G, H61P, H61W, Q62 Q62M, Q62P, Q62W, P63D, P63E, P63G, P63I, P63K, P63L, P63M, P63N, P63Q, P63R, P63S, P63T, P63V, P63W, R64D, R64E, R64F, R64L, R64M, R64P, R64Q, R64Q, R64Q, R64Q R64Y, W65A, W65E, W65G, W65K, W65L, W65M, W65N, W65P, W65R, W65V, V66C, V66G, V66I, V66M, V66N, V66Q, V66S, V66W, V66Y, L67A, L67C, L67E, L67G, L67G, L67G L67Q, L67Q, L67S, L67T, L67W, V68A, V68E, V68G, V68L, V68M, V68N, V68P, V68Q, V68S, V68T, E69A, E69C, E69D, E69F, E69G, E69H, E69K, E69L, E69M, E69M, E69M, E69M E69P, E69Q, E69S, E69V, E69W, E69Y, L70A, L70C, L70E, L70F, L70G, L70H, L70I, L70K, L70Q, L70S, L70T, L70V, L70W, G71A, G71C, G71S, G72A, G72C, G72C, G72C, G72C G72P, G72S, N73A, N73C, N73G, N73H, N73I, N73L, N73P, N73R, N73S, N73T, N73V, N73W, D74A, D74C, D74E , D74F, D74G, D74Q, D74S, D74W, D74Y, G75A, G75C, G75D, G75E, G75F, G75I, G75K, G75L, G75M, G75N, G75P, G75R, G75T, G75V, G75W, G75Y, L76A, L76C, L76C, L76C, L76C, L76C , L76E, L76F, L76G, L76I, L76K, L76M, L76N, L76P, L76Q, L76R, L76T, L76V, L76W, R77A, R77C, R77D, R77E, R77F, R77G, R77H, R77K, R77L, R77N, R77Q, R77Q, R77Q, R77Q , R77V, R77W, G78A, G78C, G78D, G78E, G78F, G78M, G78N, G78P, G78Q, G78R, G78S, G78T, G78V, G78Y, F79A, F79D, F79E, F79G, F79H, F79K, F79M, F79N, F79N, F79N , F79Q, F79S, F79V, F79W, F79Y, Q80A, Q80E, Q80G, Q80L, Q80M, Q80S, Q80W, Q80Y, P81A, P81E, P81K, P81L, P81M, P81N, P81T, P81W, P81Y, Q82A, Q82F, Q82F, Q82F, Q82F , Q82M, Q82N, Q82P, Q82R, Q82S, Q82T, Q82V, Q82W, Q82Y, Q83A, Q83C, Q83F, Q83G, Q83K, Q83L, Q83M, Q83N, Q83R, Q83S, Q83T, Q83V, Q83W, Q83Y, T84A, T84A , T84E, T84F, T84G, T84H, T84K, T84L, T84M, T84N, T84Q, T84R, T84S, T84V, T84W, T84Y, E85A, E85C, E85D, E85F, E85G, E85L, E85P, E85Q, E85R, E85S, E85S , E85V, E85W, E85Y, Q86A, Q86G, Q86H, Q86K, Q86P, Q86T, Q86V, Q86W, Q86Y, T87A, T87C, T87D, T87E, T8 7F, T87G, T87H, T87L, T87M, T87P, T87R, T87S, T87V, T87W, L88A, L88C, L88E, L88F, L88G, L88H, L88Q, L88S, L88W, L88Y, R89A, R89G, R89H, R89L, R89L, R89L, R89L R89T, R89V, R89W, Q90E, Q90L, Q90N, Q90P, Q90W, Q90Y, I91E, I91G, I91L, I91M, I91N, I91Q, I91S, I91V, I91Y, L92A, L92C, L92E, L92G, L92H, L92N, L92N, L92N, L92N L92R, L92S, L92T, L92V, L92Y, Q93A, Q93E, Q93F, Q93G, Q93H, Q93I, Q93L, Q93M, Q93N, Q93P, Q93S, Q93V, Q93W, Q93Y, D94C, D94E, D94F, D94G, D94H, D94H, D94H, D94H D94L, D94N, D94P, D94Q, D94R, D94S, D94V, V95A, V95C, V95D, V95E, V95F, V95G, V95I, V95L, V95M, V95N, V95P, V95Q, V95T, V95W, V95Y, K96A, K96C, K96C, K96C, K96C K96N, K96P, K96Q, K96R, K96V, K96Y, A97C, A97E, A97F, A97K, A97N, A97P, A97R, A97V, A97W, A98E, A98G, A98K, A98L, A98P, A98V, A98W, A98Y, N99A, N99A, N99A, N99A, N99D, N99G, N99L, N99M, N99P, N99Q, N99R, N99S, N99W, N99Y, A100D, A100E, A100G, A100H, A100I, A100K, A100L, A100M, A100Q, A100R, A100S, A100T, A100V, A100W, A100W, A100W, A100W, A100W E101A, E101D, E101G, E101L, E101M, E101P, E101S, E101T, E101V, P102E, P102F, P102G, P10 2H, P102I, P102L, P102Q, P102R, P102S, P102V, P102W, P102Y, L103A, L103C, L103E, L103G, L103I, L103K, L103N, L103Q, L103R, L103S, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T, L103T L104G, L104I, L104N, L104P, L104Q, L104S, L104W, L104Y, M105A, M105C, M105E, M105F, M105G, M105I, M105K, M105L, M105P, M105T, M105V, M105W, Q106A, Q106C, Q106D, Q106G, Q106H, Q106K, Q106L, Q106M, Q106R, Q106S, Q106T, Q106V, Q106W, Q106Y, I107A, I107C, I107E, I107F, I107G, I107K, I107L, I107M, I107Q, I107C, R107, R10, R107, R107 R108E, R108F, R108G, R108H, R108I, R108L, R108M, R108S, R108V, R108W, R108Y, L109A, L109C, L109D, L109E, L109F, L109G, L109K, L109M, L10910, L109M, L109, L109, L109, L109, L109, L109, L109, L109, L109, L109, L109, L9, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 10, 9 L109Y, P110A, P110C, P110D, P110E, P110F, P110G, P110H, P110K, P110L, P110m, P110N, P110R, P110S, P110V, P110W, A111C, A111E, A111L, A111M, A111N, A111P, A111Q, A111R, A111S, A111V, A111W, A111Y, N112A, N112F, N112G, N112I, N112K, N112L, N112P, N112R, N112V, N112W, N112Y, Y113A, Y113C, Y113D, Y113E, Y1 13G, Y113I, Y113M, Y113P, Y113Q, Y113S, Y113S, Y113W, G114A, G114F, G114K, G114L, G114M, G114P, G114W, G114Y, R115A, R115C, R115E, R115G, R115G, R115G, R115G, R115G, R115G, R115G, R115G, R115G, R115G R115V, R115W, R115Y, R116C, R116D, R116E, R116H, R116T, R116V, R116W, Y117A, Y117C, Y117D, Y117E, Y117G, Y117H, Y117I, Y117L, Y117, 7, 7, 7, 7, 7, 7, 7, 7, 7 Y117V, Y117W, N118A, N118C, N118E, N118F, N118G, N118H, N118I, N118K, N118L, N118M, N118P, N118Q, N118S, N118T, N118V, N118W, E119C, E11911, E11911, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119, E119 E119P, E119Q, E119R, E119T, E119W, E119Y, A120D, A120E, A120G, A120I, A120L, A120P, A120T, A120W, F121A, F121C, F121D, F121E, F121G, F121K, F121L, F121M, F121N, F121P, F121Q, F121R, F121S, F121V, F121W, F121Y, S122A, S122C, S122D, S122E, S122F, S122G, S122I, S122L, S122M, S122P, S122R, S122V, S122W, S122YE123, A123, A123, A123, A123, A123, 123, 123, 123 A123T, A123V, A123W, A123Y, I124A, I124C, I124D, I124E, I124G, I124H, I124K, I124L, I124R, I124S, I124T, I124W, I124Y, Y125C, Y 125F, Y125G, Y125H, Y125I, Y125L, Y125P, Y125Q, Y125R, Y125S, Y125T, Y125V, Y125W, P126C, P126F, P126H, P126K, P126R, P126T, P126V, P126Y, P126Y, K127, P7 L128C, L128E, L128F, L128G, L128Q, L128R, L128S, L128T, L128V, L128W, A129D, A129F, A129H, A129I, A129K, A129L, A129N, A129W, A129Y, K130E, K130I, K130P, K130V, E131A, E131C, E131D, E131F, E131G, E131I, E131K, E131L, E131N, E131P, E131V, E131W, F132C, F132D, F132E, F132K, F132L, F132N, F132P, F132T, F132V, D133C, D133K, D133R, D133S, D133T, D133V, D133Y, V134C, V134D, V134E, V134I, V134K, V134M, V134N, V134P, V134Q, V134R, V134S, V134W, V134Y, P135A, P135E, P135K, P135Q, L136A, L136C, L136D, L136E, L136F, L136G, L136H, L136K, L136M, L136N, L136P, L136Q, L136R, L136S, L136T, L137A, L137C, L137D, L137E, L137G, L137H, L137K, L137P, L137Q, L137R, L137S, L137Y, P138E, P138F, P138G, P138N, P138R, P138T, P138V, F139A, F139C, F139D, F139E, F139G, F139H, F139L, F139M, F139N, F139S, F139T, F139V, F139W, F140A, F140C, F140G, F140I, F140L, F140M, F140N, F140P, F140S, F140T, F140V, F140W, M141A, M141C, M141D, M141E, M141F, M141G, M141K, M141L, M141P, M141Q, M141R, M141T, M141T, M141T, M141T, M141T, M141T, M141T, M141T, M141T, M141T, M141T, M141T, M141T, M141T, M141T, M141T, M141T, M141T, M141T E142C, E142G, E142I, E142L, E142M, E142N, E142P, E142Q, E142R, E142S, E142T, E142V, E142W, E142Y, E143A, E143D, E143F, E143G, E143D, E143D, E143D, E143D, V143 V144G, V144H, V144N, V144P, V144Q, V144R, V144S, V144W, V144Y, Y145A, Y145C, Y145D, Y145E, Y145G, Y145I, Y145L, Y145M, Y145N, Y145Q, Y145R, Y145S, Y145T, Y145W, L146A, L146C, L146D, L146E, L146G, L146H, L146P, L146S, L146W, K147G, K147P, K147R, K147W, P148D, P148E, P148W, Q149L, W150C, W150D, W150E, W150G, W150L, W150P, W150Q, W150R, W150T, M150V, M151A, M151C, M151D, M151E, M151F, M151G, M151i, M151L, M151Q, M151R, M151S, M151T, M151V, M151W, Q152A, Q152D, Q152E, Q152F, Q152H, Q152I, Q152K, Q152L, Q152N, Q152P, Q152R, Q152S, Q152T, Q152V, Q152Y, D153A, D153E, D153F, D153I, D153K, D153M, D153P, D153Q, D153V, D153W, D154A, D154C, D154E, D154F, D154G, D154H, D154I, D154K, D154L, D154M, D154N, D154P, D154R, D154S, D154T, D154V, D154W, G155A, G155F, G155H, G155I, G155P, G155V, G155W, G155Y, I156A, I156C, I156E, I156F, I156G, I156K, I156L, I156M, I156Q15 H157E, P158A, P158F, P158G, P158H, P158I, P158L, P158Q, P158S, P158T, P158V, P158W, N159C, N159E, N159G, N159I, N159K, N159L, N159M, N159P, N159Q, N159R, N159T, N159V, N159W, R160A, R160C, R160D, R160E, R160G, R160H, R160I, R160K, R160N, R160Q, R160S, R160W, D161E, D161G, D161I, D161K, D161L, D161M, D161N, D161Q, D161R, D161S, D161V, D161W, A162G, A162I, A162K, A162L, A162N, A162R, A162T, A162V, A162Y, Q163A, Q163C, Q163D, Q163E, Q163F, Q163G, Q163I, Q163L, Q163M, Q163S, Q163T, Q163V, Q163W, Q163Y, P164A, P164C, P164D, P164K, P164L, P164M, P164N, P164R, P164T, P164V, P164W, F165D, F165E, F165G, F165H, F165I, F165K, F165L, F165M, F165R, F165S, F165T, F165V, F165W, F165Y, I166A, I166C, I166F, I166L, I166M, I166S, I166V, I166Y, A167C, A167D, A167E, A167F, A167G, A167K, A167L, A167M, A167N, A167Q, A167R, A167T, A167V, A167W, A167Y, D168A, D168G, D168H, D168L, D168M, D168P, D168R, D168T, D168V, D168W, W169A, W169D, W169E, W169G, W169K, W169M, W169Q, W169R, W169S, W169T, W169V, M170A, M170E, M170F, M170G, M170H, M170L, M170N, M170Q, M170S, M170T, M170V, M170W, M170Y, A171E, A171F, A171I, A171S, A171V, A171W, K172A, K172M, K172, Q172, K172 Q173Y, L174A, L174F, L174G, L174Q, L174S, L174T, L174W, L174W, L174Y, Q175F, Q175I, Q175L, Q175M, Q175Y, P176D, P176H, P176K , P176L, P176N, P176Q, P176R, P176V, P176W, P176Y, L177D, L177F, L177G, L177M, L177S, L177T, V178A, V178F, V178G, V178K, V178L, V178R, V178S, V178T, V178W, N179G, N179H, N179R , N179T, N179V, N179W, N179Y, H180A, H180E, H180G, H180L, H180P, H180R, H180S, H180V, H180W, D181A, D181C, D181E, D181G, D181H, D181I, D181L, D181P, D181Q, D181R, D181S, D181T , D181W, S182A, S182C, S182D, S182E, S182G, S182I, S182K, S182L, S182N, S182P, S182Q, S182R, S182T and / or S180V, where, in the replacement mutation designations, the numbers refer to positions of amino acids of SEQ ID NO: 31.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que tiene una especificidad por el sustrato, y/o una actividad del sustrato, aumentada (por ejemplo, velocidad catalitica), con respecto a sustratos de C10 (es decir, sustratos, cuyas cadenas de carbono tienen una longitud de 10 carbonos), y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a uno o mas de los restos seleccionados de 5-30, 35-60, 65-98, 102-139 y/o 140-180 de la SEC ID NO: 31. El aumento de la especificidad por el sustrato y/o de la actividad del sustrato con respecto a sustratos de C10 puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which has a specificity for the substrate, and / or an activity of the substrate, increased (for example, catalytic velocity), with respect to C10 substrates ( that is, substrates, whose carbon chains have a length of 10 carbons), and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated in one or more amino acid positions corresponding to one or more of the residues selected from 5-30, 35-60, 65-98, 102-139 and / or 140-180 of the SEC ID NO: 31. The increase in substrate specificity and / or substrate activity with respect to C10 substrates can be measured in vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que tiene una especificidad por el sustrato, y/o una actividad del sustrato, aumentada (por ejemplo, velocidad catalitica), con respecto a sustratos de C10 y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a uno o mas restos de la SEC ID NO: 31 seleccionados de 1, 3, 4, 7, 9, 12, 13, 14, 16, 17, 20, 22, 24, 25, 28, 32, 38, 39, 40, 42, 43, 46, 47, 48, 49, 50, 51, 52, 54, 56, 59, 60, 64, 68, 72, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 89, 90, 91, 92, 93, 94, 95, 96, 98, 99, 100, 101, 102, 103, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 130 ,132, 133, 134, 138, 139, 140, 141, 142, 144, 145, 146, 147, 148, 150, 151, 152, 156, 158, 159, 160, 161, 162, 163, 164, 165 , 166, 167, 168, 169, 170, 171, 172, 173, 175, 176, 177, 178, 179, 180, 181 y/o 182. El aumento de la especificidad por el sustrato y/o de la actividad del sustrato con respecto a sustratos de C10 puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which has a specificity for the substrate, and / or an activity of the substrate, increased (eg, catalytic velocity), with respect to C10 substrates and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated in one or more amino acid positions corresponding to one or more residues of SEQ ID NO: 31 selected from 1, 3, 4, 7, 9, 12, 13, 14, 16, 17 , 20, 22, 24, 25, 28, 32, 38, 39, 40, 42, 43, 46, 47, 48, 49, 50, 51, 52, 54, 56, 59, 60, 64, 68, 72 , 75, 76, 77, 78, 79, 80, 81.82, 83, 84, 85, 86, 87, 89, 90, 91, 92, 93, 94, 95, 96, 98, 99, 100, 101 , 102, 103, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127 , 128, 130, 132, 133, 134, 138, 139, 140, 141, 142, 144, 145, 146, 147, 148, 150, 151, 152, 156, 158, 159, 160, 161, 162, 163 , 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 175, 176, 177, 178, 179, 180, 181 and / or 182. The increase in specificity for the substrate and / or The substrate activity with respect to C10 substrates can be measured in vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o un equivalente natural), que tiene una especificidad por el sustrato, y/o una actividad del sustrato, aumentada (por ejemplo, velocidad catalitica), con respecto a sustratos de C10 y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta con una o mas sustituciones seleccionadas de A1L, A1S, T3K, L4A, L7M, L7V, D9N, S12A, A13D, G14A, G14E, G14P, G14Q, G14R, G14S, G14V, R16G, R16L, R16M, R16N, R16P, R16Q, R16T, M17C, M17L, M17T, M17V, S20A, S20C, S20D, S20G, S20L, S20T, S20W, A22C, A22D, A22E, A22G, A22H, A22I, A22K, A22N, P24A, P24C, P24D, P24F, P24I, P24S, P24T, P24V, P24W, A25E, A25L, A25N, A25Q, A25V, N28A, N28R, Q32V, Q32Y, V38E, V38K, V38R, N39A, N39T, A40D, A40H, I42A, I42E, I42L, I42S, I42T, I42W, I42Y, S43A, S43C, S43D, S43E, S43L, S43N, S43P, T46E, T46F, T46I, T46L, T46V, S47A, S47C, S47F, S47G, S47L, S47M, S47T, S47V, Q48D, Q48E, Q48G, Q48S, Q48T, Q48V, Q48W, Q49A, Q49C, Q49D, Q49G, Q49H, Q49L, Q49M, Q49S, G50A, G50Q, L51A, L51F, L51H, L51Y, A52D, A52M, L54T, A56P, K59R, Q60M, R64D, R64E, R64Q, V68L, G72A, G72C, G72P, G72S, G75A, G75C, G75D, G75E, G75F, G75I, G75K, G75L, G75M, G75N, G75P, G75T, G75V, G75W, G75Y, L76A, L76D, L76G, L76I, L76K, L76M, L76N, L76P, L76Q, L76R, L76W, R77G, R77L, R77Q, G78A, G78C, G78E, G78F, G78M, G78N, G78Q, G78R, G78S, G78T, G78V, G78Y, F79A, F79D, F79E, F79G, F79H, F79N, F79Q, F79W, F79Y, Q80E, P81N, P81T, P81Y, Q82R, Q82S, Q82T, Q83A, Q83C, Q83F, Q83G, Q83K, Q83L, Q83M, Q83N, Q83R, Q83S, Q83T, Q83V, Q83W, Q83Y, T84A, T84F, T84L, T84M, T84N, T84Q, T84V, T84Y, E85A, E85C, E85L, E85Q, E85R, E85S, E85T, E85W, E85Y, Q86A, Q86G, Q86K, Q86T, T87D, T87P, R89A, R89G, Q90E, Q90Y, I91V, L92V, Q93A, Q93E, Q93G, Q93H, Q93I, Q93L, Q93S, Q93W, Q93Y, D94E, D94F, D94G, D94H, D94K, D94N, D94Q, D94R, D94S, D94V, V95L, V95T, K96V, K96Y, A98W, N99G, N99L, N99P, N99Q, N99R, N99Y, A100G, A100V, E101A, E101D, E101G, E101L, E101M, E101S, E101T, E101V, P102S, L103G, M105C, M105I, M105V, Q106A, Q106D, Q106H, Q106W, I107Y, R108A, R108D, R108E, R108F, R108G, R108H, R108I, R108L, R108M, R108S, R108W, R108Y, L109A, L109D, L109E, L109F, L109G, L109K, L109P, L109R, L109S, L109Y, P110C, P110D, P110E, P110F, P110G, P110H, P110K, P110L, P110M, P110N, P110R, P110S, P110V, P110W, A111C, A111E, A111L, A111M, A111P, A111Q, A111R, A111V, A111W, A111Y, N112A, N112F, N112G, N112K, N112R, N112W, Y113A, Y113C, Y113G, Y113I, Y113M, G114K, G114L, G114P, R115A, R115C, R115E, R115G, R115N, R115S, R115W, R115Y, R116D, R116E, R116W, Y117A, Y117C, Y117E, Y117I, Y117L, Y117N, Y117Q, Y117R, Y117S, Y117T, Y117V, N118C, N118G, N118I, N118K, N118S, N118T, N118V, N118W, E119C, E119F, E119G, E119K, E119M, E119R, E119W, E119Y, A120D, A120E, A120G, A120W, F121A, F121D, F121E, F121M, F121P, F121Q, F121R, F121S, F121Y, S122D, S122E, S122F, S122I, S122L, S122M, S122V, S122W, S122Y, A123H, A123L, A123V, I124T, Y125C, Y125F, Y125G, Y125P, Y125S, Y125V, Y125W, P126R, P126T, P126V, P126Y, K127S, L128C, L128T, L128V, K130E, K130I, K130V, F132D, F132E, F132N, F132T, D133K, D133R, D133S, D133T, D133V, D133Y, V134I, V134M, V134S, P138E, P138N, P138R, P138T, P138V, F139A, F139D, F139G, F139H, F139M, F139S, F139W, F140C, F140G, F140M, F140N, F140P, F140S, M141A, M141C, M141D, M141E, M141F, M141G, M141K, M141L, M141P, M141Q, M141R, M141T, M141V, M141W, M141Y, E142A, E142C, E142P, E142Q, E142W, E142Y, V144D, V144E, V144G, V144H, V144N, V144P, V144Q, V144R, V144S, V144W, V144Y, Y145A, Y145C, Y145D, Y145E, Y145G, Y145I, Y145L, Y145M, Y145N, Y145Q, Y145T, Y145W, L146A, L146C, L146D, L146E, L146G, L146H, L146S, L146W, K147G, K147P, K147W, P148D, P148E, W150C, W150D, W150E, W150G, W150L, W150Q, W150T, M151A, M151C, M151E, M151F, M151G, M151I, M151Q, M151S, M151T, M151V, M151W, Q152D, Q152F, Q152I, Q152L, Q152T, I156L, P158A, P158F, P158G, P158H, P158I, P158L, P158Q, P158T, P158V, N159C, N159E, N159G, N159I, N159K, N159L, N159M, N159R, N159T, N159V, R160A, R160C, R160D, R160E, R160G, R160H, R160N, R160Q, R160S, R160W, D161E, D161G, D161I, D161K, D161L, D161M, D161Q, D161R, D161W, A162I, A162L, A162T, A162V, A162Y, Q163G, Q163L, Q163M, Q163S, P164A, P164C, P164D, P164M, P164N, P164R, P164V, P164W, F165D, F165E, F165G, F165H, F165I, F165K, F165L, F165M, F165R, F165S, F165T, F165V, F165Y, I166F, I166L, I166M, I166V, A167C, A167M, A167R, A167T, D168G, D168P, D168R, W169E, W169K, W169Q, M170F, M170H, M170L, M170T, M170V, M170Y, A171E, A171F, A171V, A171W, K172A, K172M, Q173N, Q175I, P176H, P176K, P176N, P176W, L177M, L177T, V178T, V178W, N179G, N179H, N179R, N179T, N179V, N179Y, H180E, H180G, H180R, H180V, H180W, D181A, D181H, D181I, D181L, D181P, D181R, D181W, S182A, S182G, S182K, S182L, S182P y/o S182R, en donde, en las designaciones de mutacion por sustitucion, los numeros se refieren a posiciones de aminoacidos de la SEC ID NO: 31. El aumento de la especificidad por el sustrato y/o de la actividad del sustrato con respecto a sustratos de C10 puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or a natural equivalent) is disclosed, which has a specificity for the substrate, and / or an activity of the substrate, increased (eg, catalytic velocity), with respect to C10 substrates and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated with one or more substitutions selected from A1L, A1S, T3K, L4A, L7M, L7V, D9N, S12A, A13D, G14A, G14E, G14P, G14Q, G14R, G14S, G14V, R16G, R16L, R16M, R16N, R16P, R16Q, R16T, M17C, M17L, M17T, M17V, S20A, S20C, S20D, S20G, S20L, S20T, S20W, A22C, A22D, A22E, A22G, A22H, A22I, A22K, A22K, A22K, A22K, A22K, A22K P24A, P24C, P24D, P24F, P24I, P24S, P24T, P24V, P24W, A25E, A25L, A25N, A25Q, A25V, N28A, N28R, Q32V, Q32Y, V38E, V38K, V38R, N39A, N39TH, A40D, A40D, A40D I42A, I42E, I42L, I42S, I42T, I42W, I42Y, S43A, S43C, S43D, S43E, S43L, S43N, S43P, T46E, T46F, T46I, T46L, T46V, S47A, S47C, S47F, S47G, S47L, S47L, S47L, S47L, S47L, S47L S47T, S47V, Q48D, Q48E, Q48G, Q48S, Q48T, Q48V, Q48W, Q49A, Q49C, Q49D, Q49G, Q49H, Q49L, Q49M, Q49S, G50A, G50Q, L51A, L51F, L51H, L51YM, A52D, A52D, A52D L54T, A56P, K59R, Q60M, R64D, R64E, R64Q, V68L, G72A, G72C, G72P, G72S, G75A, G75C, G75D, G75E, G75F, G75I, G75K, G75L, G75M, G75N, G75P, G75T, G75T, G75T, G75T, G75T G75W, G75Y, L76A, L76D, L76G, L76I, L76K, L76M, L76N, L76P, L7 6Q, L76R, L76W, R77G, R77L, R77Q, G78A, G78C, G78E, G78F, G78M, G78N, G78Q, G78R, G78S, G78T, G78V, G78Y, F79A, F79D, F79E, F79G, F79Q, F79N, F79N, F79N, F79N F79W, F79Y, Q80E, P81N, P81T, P81Y, Q82R, Q82S, Q82T, Q83A, Q83C, Q83F, Q83G, Q83K, Q83L, Q83M, Q83N, Q83R, Q83S, Q83T, Q83V, Q83W, Q83Y, T84A, T84F, T84L, T84M, T84N, T84Q, T84V, T84Y, E85A, E85C, E85L, E85Q, E85R, E85S, E85T, E85W, E85Y, Q86A, Q86G, Q86K, Q86T, T87D, T87P, R89A, R89G, Q90E, R89G, Q90 Q90Y, I91V, L92V, Q93A, Q93E, Q93G, Q93H, Q93I, Q93L, Q93S, Q93W, Q93Y, D94E, D94F, D94G, D94H, D94K, D94N, D94Q, D94R, D94S, D94V, V95L, V95T, K95 K96Y, A98W, N99G, N99L, N99P, N99Q, N99R, N99Y, A100G, A100V, E101A, E101D, E101G, E101L, E101M, E101S, E101T, E101V, P102S, L103G, M10510, M10510, M105D, M10510, M105D, M10510 Q106H, Q106W, I107Y, R108A, R108D, R108E, R108F, R108G, R108H, R108I, R108L, R108M, R108S, R108W, R108Y, L109A, L109D, L109E, L109F, L109G, L109K, L109P, L109R, L109S, L109Y, P110C, P110D, P110E, P110F, P110G, P110H, P110K, P110L, P110m, P110N, P110R, P110S, P110V, P110W, A111C, A111E, A111L, A111M, A111P, A111Q, A111R, A111V, A111W, A111Y, N112A, N112F, N112G, N112K, N112R, N112W, Y113A, Y113C, Y113G, Y113I, Y113M, G114K, G114L, G114P, R115A, R115C, R115E, R115G, R115N, R115S, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E, R115E6 Y117C, Y 117E, Y117I, Y117L, Y117N, Y117Q, Y117R, Y117S, Y117T, Y117V, N118C, N118G, N118I, N118K, N118S, N118T, N118V, N118W, E119C, E119F, E119G, E119G, E119G, E119G A120D, A120E, A120G, A120W, F121A, F121D, F121E, F121M, F121P, F121Q, F121R, F121S, F121Y, S122D, S122E, S122F, S122I, S122L, S122M, S12WV, S122V, S122V, S122V, S122V, S122V, S122V123 I124T, Y125C, Y125F, Y125G, Y125P, Y125S, Y125V, Y125W, P126R, P126T, P126V, P126Y, K127S, L128C, L128T, L128V, K130E, K130I, K130V, F132D, F132E, F132N, F132T, D133K, D133R, D133S, D133T, D133V, D133Y, V134I, V134M, V134S, P138E, P138N, P138R, P138T, P138V, F139A, F139D, F139G, F139H, F139M, F139S, F13940, F140, F140, F140, F140, F140, F140 M141A, M141C, M141D, M141E, M141F, M141G, M141K, M141L, M141P, M141Q, M141R, M141T, M141V, M141W, M141Y, E142A, E142C, E142P, E142Q, E142Q, V142, V142, V142, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V14, V142 V144N, V144P, V144Q, V144R, V144S, V144W, V144Y, Y145A, Y145C, Y145D, Y145E, Y145G, Y145I, Y145L, Y145M, Y145N, Y145Q, Y145T, Y145W, L146A, L146C, L146D, L146E, L146G, L146H, L146S, L146W, K147G, K147P, K147W, P148D, P148E, W150C, W150D, W150E, W150G, W150L, W150Q, W150T, M151A, M151C, M151E, M151F, M151G, M151I, M151Q, M151S, M151T, M151V, M151W, Q152D, Q152F, Q152I, Q152L, Q152T, I156L, P158A, P158F, P158G, P158H, P158I, P158L, P15815, P15815, P15815, P15815, N159 N159I, N159K, N159L, N159M, N159R, N159T, N159V, R160A, R 160c, R 160d, R 160e, R160G, R160H, R160N, R160Q, R160S, R160W, D161E, D161G, D161I, D161K, D161L, D161M, D161Q, D161R, D161W, A162I, A162L, A162T, A162V, A162Y, Q163G, Q163L, Q163M, Q163S, P164A, P164C, P164D, P164M, P164N, P164R, P164V, P164W, F165D, F165E, F165G, F165H, F165I, F165K, F165L, F165M, F165R, F165S, F165T, F165V, F165Y, I166F, I166L, I166M, I166V, A167C, A167M, A167R, A167T, D168G, D168P, D168R, W169E, W169K, W169Q, M170F, M170H, M170L, M170T, M170V, M170Y, A171E, A171F, A171V, A171W, K172A, K172M, Q173N, Q175I, P176H, P176K, P176N, P176W, L177M, L177T, V178T, V178W, N179G, N179H, N179R, N179T, N179V, N179Y, H180E, H180G, H180R, H180V, H180W, D181A, D181H, D181I, D181L, D181P, D181R, D181W, S182A, S182 G18 , in substitution mutation designations, the numbers refer to amino acid positions of SEQ ID NO: 31. The increase in substrate specificity and / or substrate activity with respect to C10 substrates can be measured in In vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que tiene una especificidad por el sustrato, y/o una actividad de sustrato, aumentada (por ejemplo, velocidad catalitica), con respecto a sustratos de C¹² (es decir, sustratos, cuyas cadenas de carbono tienen una longitud de 12 carbonos), y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a los restos 10-25, 35-85, 90-103, 110-143, 146-180 de la SEC ID NO: 31. El aumento de la especificidad por el sustrato y/o de la actividad del sustrato con respecto a sustratos de C¹² puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which has a specificity for the substrate, and / or an increased substrate activity (eg, catalytic velocity), with respect to C ¹² substrates (i.e. substrates, whose carbon chains have a length of 12 carbons), and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated in one or more amino acid positions corresponding to residues 10-25, 35-85, 90-103, 110-143, 146-180 of SEQ ID NO: 31. The increase in the specificity for the substrate and / or the activity of the substrate with respect to C ¹² substrates can be measured in vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o un equivalente natural), que tiene una especificidad por el sustrato, y/o una actividad del sustrato, aumentada (por ejemplo, velocidad catalitica), con respecto a sustratos de C12 y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a uno o mas restos de la SEC ID NO: 31 seleccionados de 1, 2, 3, 4, 5, 6, 7, 9, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 24, 25, 26, 27, 28, 29, 30, 31, 35, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 57, 58, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 76, 77, 78, 79, 80, 81, 82, 83, 84, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 111, 112, 113, 114, 115, 116, 117, 119, 120, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 133, 134, 136, 137, 140, 141, 142, 145, 149, 152, 153, 155, 156, 158, 159, 160, 161, 162, 163, 164, 166, 167, 168, 169, 170, 172, 173, 174, 175, 176, 177, 179, 180, 181 y/o 182. El aumento de la especificidad por el sustrato y/o de la actividad del sustrato con respecto a sustratos de C12 puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or a natural equivalent) is disclosed, which has a specificity for the substrate, and / or an activity of the substrate, increased (eg, catalytic velocity), with respect to C12 substrates and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated in one or more amino acid positions corresponding to one or more residues of SEQ ID NO: 31 selected from 1, 2, 3, 4, 5, 6, 7, 9, 11, 12 , 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 24, 25, 26, 27, 28, 29, 30, 31, 35, 37, 38, 39, 40, 41, 42 , 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 57, 58, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69 , 70, 71, 72, 73, 74, 76, 77, 78, 79, 80, 81, 82, 83, 84, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96 , 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 111, 112, 113, 114, 115, 116, 117, 119, 120, 122, 123, 124 , 125, 126, 127, 128, 129, 130, 131, 133, 134, 136, 137, 140, 141, 142, 145, 149, 152, 153, 155, 156, 158, 159, 160, 161, 162 , 163, 164, 166, 167, 168, 169, 170, 172, 173, 174, 175, 176, 177, 179, 180, 181 and / or 182. The increase in specificity for the substrate and / or the Substrate activity with respect to C12 substrates can be measured in vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que tiene una especificidad por el sustrato, y/o una actividad del sustrato, aumentada (por ejemplo, velocidad catalitica), con respecto a sustratos de C12 y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta con una o mas sustituciones seleccionadas de A1Q, A1S, A1V, D2E, D2K, D2P, D2W, T3R, T3W, L4A, L4Y, L5F, L5G, L5S, L5Y, I6T, I6V, L7A, L7C, L7M, L7N, L7S, L7T, L7V, L7Y, D9N, L11M, S12A, S12I, S12V, A13C, A13G, A13H, A13I, A13L, A13N, A13T, A13W, G14F, G14I, G14K, G14M, G14V, Y15A, Y15C, Y15D, Y15E, Y15G, Y15I, Y15L, Y15M, Y15N, Y15Q, Y15R, Y15S, Y15V, R16D, R16E, R16G, R16H, R16I, R16L, R16N, R16P, R16S, R16T, R16V, R16W, M17A, M17C, M17G, M17K, M17N, M17P, M17Q, M17R, M17S, M17T, S18M, S18N, A19L, S20A, S20C, S20D, S20G, S20L, S20T, S20W, A21I, A21L, A21P, A21Y, A22F, A22L, A22M, A22N, A22R, A22Y, P24G, P24V, A25D, A25E, A25L, A25N, A25Q, A25R, A25V, L26D, L26E, L26F, L26G, L26H, L26I, L26K, L26N, L26R, L26S, L26W, L26Y, L27A, L27C, L27F, L27M, L27W, L27Y, N28R, N28W, D29P, K30P, W31E, W31N, T35L, T35Y, V37F, V37S, V37W, V38D, V38F, V38G, V38P, N39A, N39C, N39E, N39G, N39Q, N39W, A40D, A40L, A40M, A40P, A40V, A40Y, S41C, S41T, I42A, I42C, I42D, I42E, I42G, I42K, I42L, I42M, I42P, I42S, I42T, I42W, I42Y, S43A, S43D, S43E, S43F, S43G, S43H, S43L, S43M, S43N, S43R, S43T, S43V, G44C, G44E, G44H, G44K, G44L, G44N, G44Q, G44R, G44S, D45A, D45C, D45E, D45F, D45H, D45I, D45K, D45L, D45M, D45P, D45Q, D45S, D45T, D45V, D45W, T46A, T46C, T46D, T46G, T46K, T46N, T46R, T46S, S47P, S47Q, Q48E, Q48V, Q48W, Q48Y, Q49A, Q49C, Q49D, Q49E, Q49G, Q49H, Q49I, Q49K, Q49L, Q49M, Q49P, Q49R, Q49S, Q49V, Q49W, Q49Y, G50A, G50C, G50F, G50I, G50K, G50L, G50M, G50N, G50P, G50Q, G50R, G50S, G50T, G50Y, L51A, L51D, L51N, L51T, L51V, L51W, A52C, A52M, A52P, A52W, R53A, R53C, R53D, R53E, R53F, R53G, R53I, R53K, R53L, R53N, R53S, R53T, R53V, R53W, R53Y, L54A, L54C, L54E, L54F, L54G, L54M, L54N, L54S, L54W, L54Y, P55Y, L57A, L57C, L57F, L57K, L57P, L57Q, L57R, L57Y, L58A, L58D, L58E, L58G, L58H, L58N, L58R, L58S, L58W, L58Y, Q60P, H61D, H61G, H61P, Q62P, Q62W, P63I, P63L, P63N, P63S, P63T, P63V, P63W, R64F, R64P, R64W, R64Y, W65A, W65E, W65G, W65K, W65M, W65N, W65V, V66M, V66S, L67A, L67T, V68A, V68L, V68M, V68S, V68T, E69A, E69C, E69D, E69G, E69H, E69K, E69L, E69M, E69N, E69P, E69V, E69Y, L70A, L70C, L70E, L70F, L70H, L70I, L70K, L70Q, L70S, L70T, L70V, G71A, G72A, N73G, N73H, N73L, N73R, N73S, N73T, D74E, D74G, L76I, L76M, L76W, R77C, R77D, R77E, R77G, R77K, R77L, R77Q, R77S, R77V, R77W, G78D, F79P, Q80G, Q80M, Q80S, Q80Y, P81A, P81E, P81K, P81L, P81M, P81W, P81Y, Q82F, Q82V, Q82W, Q82Y, Q83A, T84E, T84R, T84W, Q86A, Q86T, T87E, T87G, T87L, L88C, R89L, R89P, Q90N, Q90P, Q90W, I91G, I91M, I91S, I91V, I91Y, L92A, L92C, L92G, L92H, L92N, L92S, L92T, L92V, L92Y, Q93A, Q93G, Q93H, Q93I, Q93P, Q93Y, D94P, V95F, V95G, V95L, V95N, V95Q, V95T, V95W, K96A, K96L, K96P, K96Y, A97K, A97P, A98L, A98P, A98V, A98W, A98Y, N99C, N99D, N99G, N99L, N99M, N99P, N99Q, N99R, N99W, N99Y, A100D, A100E, A100G, A100H, A1001, A100K, A100L, A100Q, A100R, A100V, A100W, A100Y, E101G, E101L, E101M, E101P, E101S, E101T, E101V, P102E, P102F, P102H, P102L, P102Q, P102R, P102S, P102W, P102Y, L103E, L103K, L103N, L103Q, L103R, L104C, L104P, L104S, L104W, M105C, M105E, M105G, M105V, Q106A, Q106C, Q106G, Q106K, Q106R, Q106S, Q106T, I107C, I107E, I107K, I107L, I107M, I107S, I107V, R108F, R108W, L109M, A111C, A111Q, A111W, N112A, N112G, N112W, Y113A, Y113D, Y113G, Y113I, G114K, G114L, G114M, G114Y, R115A, R115C, R115E, R115G, R115N, R115S, R115Y, R116H, R116W, Y117C, Y117H, Y117I, Y117L, Y117M, Y117N, Y117S, Y117T, Y117V, E119C, E119F, E119K, E119M, E119R, E119W, E119Y, A120D, A120G, A120I, A120T, A120W, S122F, S122I, S122L, S122M, S122V, S122W, S122Y, A123C, A123F, A123H, A123L, A123R, A123T, A123V, A123W, A123Y, I124G, I124H, I124K, I124L, I124R, I124S, I124Y, Y125F, Y125R, P126C, P126F, P126H, P126Y, K127I, K127P, L128A, L128S, L128T, A129H, A129I, A129K, A129N, A129W, A129Y, K130P, E131A, E131C, E131F, E131G, E131K, E131L, E131N, E131V, E131W, D133K, V134D, V134E, V134K, V134N, V134Q, V134R, V134W, V134Y, L136A, L136D, L136E, L136F, L136G, L136H, L136K, L136N, L136P, L136Q, L136R, L136S, L136T, L137E, L137G, L137H, L137P, L137Q, L137S, L137Y, F140M, M141A, M141C, M141L, M141P, E142C, Y145E, Q149L, Q152A, Q152D, Q152E, Q152H, Q152K, Q152R, Q152Y, D153K, G155F, G155W, G155Y, I156C, I156F, I156M, I156V, P158A, P158G, N159G, N159Q, N159T, N159V, R160A, R160D, R160E, R160G, R160H, R160N, R160Q, R160S, R160W, D161I, D161K, D161L, D161M, D161N, D161Q, D161W, A162G, Q163A, Q163C, Q163G, Q163L, Q163M, Q163S, Q163T, P164C, P164M, I166L, I166V, A167C, A167E, A167F, A167G, A167K, A167L, A167N, A167Q, A167R, A167T, A167V, A167Y, D168G, D168H, D168L, D168R, D168V, D168W, W169A, W169D, W169E, W169G, W169K, W169Q, W169S, M170F, M170G, M170N, M170Q, M170S, M170V, M170W, K172M, K172P, Q173N, L174A, L174F, L174G, L174T, L174W, Q175I, P176H, P176K, P176L, P176N, P176W, L177D, L177G, N179H, N179R, N179Y, H180A, H180G, D181H, D181I, D181L, D181R, D181W, S182K, S182L, S182P y/o S182R, en donde, en las designaciones de mutacion por sustitucion, los numeros se refieren a posiciones de aminoacidos de la SEC ID NO: 31. El aumento de la especificidad por el sustrato y/o de la actividad del sustrato con respecto a sustratos de C12 puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which has a specificity for the substrate, and / or an activity of the substrate, increased (eg, catalytic velocity), with respect to C12 substrates and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated with one or more substitutions selected from A1Q, A1S, A1V, D2E, D2K, D2P, D2W, T3R, T3W, L4A, L4Y, L5F, L5G, L5S, L5Y, I6T, I6V, L7A, L7C, L7M, L7N, L7S, L7T, L7V, L7Y, D9N, L11M, S12A, S12I, S12V, A13C, A13G, A13H, A13I, A13L, A13N, A13T, A13W, G14F, G14I, G14K, G14K G14V, Y15A, Y15C, Y15D, Y15E, Y15G, Y15I, Y15L, Y15M, Y15N, Y15Q, Y15R, Y15S, Y15V, R16D, R16E, R16G, R16H, R16I, R16L, R16N, R16P, R16S, R16T, R16T, R16T, R16T, R16T R16W, M17A, M17C, M17G, M17K, M17N, M17P, M17Q, M17R, M17S, M17T, S18M, S18N, A19L, S20A, S20C, S20D, S20G, S20L, S20T, S20W, A21I, A21L, A21P, A21Y, A22F, A22F, A22F, A22F, A22F, A22F, A22F, A22F A22M, A22N, A22R, A22Y, P24G, P24V, A25D, A25E, A25L, A25N, A25Q, A25R, A25V, L26D, L26E, L26F, L26G, L26H, L26I, L26K, L26N, L26R, L26S, L26W, L26W, L26W L27A, L27C, L27F, L27M, L27W, L27Y, N28R, N28W, D29P, K30P, W31E, W31N, T35L, T35Y, V37F, V37S, V37W, V38D, V38F, V38G, V38P, N39A, N39C, N39E, N39E, N39E, N39E, N39Q, N39W, A40D, A40L, A40M, A40P, A40V, A40Y, S41C, S41T, I42A, I42C, I42D, I42E, I42G, I42K, I42L, I42M, I42P, I42S, I42T, I42W, I42Y, S43A, S43A, S43A, S43A, S43A, S43A, S43A S43E, S43F, S43G, S43H, S43L, S43M, S43N, S43R, S43T, S43V, G44C, G44E, G44H, G44K, G44L, G44N, G44Q, G44R, G44S, D45A, D45C, D45E, D45F, D45H, D45H D45K, D45L, D45M, D45P, D45Q, D45S, D45T, D45V, D45W, T46A, T46C, T46D, T46G, T46K, T46N, T46R, T46S, S47P, S47Q, Q48E, Q48V, Q48W, Q48Y, Q49A, Q49A, Q49A, Q49A, Q49A, Q49A, Q49A Q49D, Q49E, Q49G, Q49H, Q49I, Q49K, Q49L, Q49M, Q49P, Q49R, Q49S, Q49V, Q49W, Q49Y, G50A, G50C, G50F , G50I, G50K, G50L, G50M, G50N, G50P, G50Q, G50R, G50S, G50T, G50Y, L51A, L51D, L51N, L51T, L51V, L51W, A52C, A52M, A52P, A52W, R53A, R53C, R53D, R53D, R53D , R53F, R53G, R53I, R53K, R53L, R53N, R53S, R53T, R53V, R53W, R53Y, L54A, L54C, L54E, L54F, L54G, L54M, L54N, L54S, L54W, L54Y, P55Y, L57A, L57C, L57C, L57C, L57C , L57K, L57P, L57Q, L57R, L57Y, L58A, L58D, L58E, L58G, L58H, L58N, L58R, L58S, L58W, L58Y, Q60P, H61D, H61G, H61P, Q62P, Q62W, P63I, P63L, P63N, P63N, P63N, P63N , P63T, P63V, P63W, R64F, R64P, R64W, R64Y, W65A, W65E, W65G, W65K, W65M, W65N, W65V, V66M, V66S, L67A, L67T, V68A, V68L, V68M, V68S, V68T, E69A, E69A, E69A , E69D, E69G, E69H, E69K, E69L, E69M, E69N, E69P, E69V, E69Y, L70A, L70C, L70E, L70F, L70H, L70I, L70K, L70Q, L70S, L70T, L70V, G71A, G72A, N73G, N73G, N73G , N73L, N73R, N73S, N73T, D74E, D74G, L76I, L76M, L76W, R77C, R77D, R77E, R77G, R77K, R77L, R77Q, R77S, R77V, R77W, G78D, F79P, Q80G, Q80M, Q80S, Q80S, Q80S, Q80S , P81A, P81E, P81K, P81L, P81M, P81W, P81Y, Q82F, Q82V, Q82W, Q82Y, Q83A, T84E, T84R, T84W, Q86A, Q8 6T, T87E, T87G, T87L, L88C, R89L, R89P, Q90N, Q90P, Q90W, I91G, I91M, I91S, I91V, I91Y, L92A, L92C, L92G, L92H, L92N, L92S, L92T, L92V, L92Y, Q93 Q93G, Q93H, Q93I, Q93P, Q93Y, D94P, V95F, V95G, V95L, V95N, V95Q, V95T, V95W, K96A, K96L, K96P, K96Y, A97K, A97P, A98L, A98P, A98V, A98W, A98Y, N99 N99D, N99G, N99L, N99M, N99P, N99Q, N99R, N99W, N99Y, A100D, A100E, A100G, A100H, A1001, A100K, A100L, A100Q, A100R, A100V, A100W, A100Y, E101G, E101L, E101M, E101M, E101M, E101M, E101M E101S, E101T, E101V, P102E, P102F, P102H, P102L, P102Q, P102R, P102S, P102W, P102Y, L103E, L103K, L103N, L103Q, L103R, L104C, L104P, L1045, L104P, M104P, M104P Q106A, Q106C, Q106G, Q106K, Q106R, Q106S, Q106T, I107C, I107E, I107K, I107L, I107M, I107S, I107V, R108F, R108W, L109M, A111C, A111Q, A111W, N112A, N112G, N112W, Y113A, Y113D, Y113G, Y113I, G114K, G114L, G114M, G114Y, R115A, R115C, R115E, R115G, R115N, R115S, R115Y, R116H, R116W, Y117C, Y117H, Y117I, Y117L, Y117M, Y117N, Y117S, Y117T, Y117V, E119C, E119F, E119 K, E119M, E119R, E119W, E119Y, A120D, A120G, A120I, A120T, A120W, S122F, S122I, S122L, S122M, S122V, S122W, S122Y, A123C, A123F, A123H, A123L, A123R, A123R, A123R, A123R, A123R, A123R, A123R, A123R, A123R, A123R, A123R, A123 R123 A123Y, I124G, I124H, I124K, I124L, I124R, I124S, I124Y, Y125F, Y125R, P126C, P126F, P126H, P126Y, K127I, K127P, L128A, L128S, L128T, A129, 12, 12, 12, 12, 12, 12, 912 K130P, E131A, E131C, E131F, E131G, E131K, E131L, E131N, E131V, E131W, D133K, V134D, V134E, V134K, V134N, V134Q, V134R, V134W, V134Y, L136A, L136D, L136E, L136F, L136G, L136H, L136K, L136N, L136P, L136Q, L136R, L136S, L136T, L137E, L137G, L137H, L137P, L137Q, L137S, L137Y, F140M, M141A, M141C, M141L, M141P, E142C, Y145E, Q149L, Q152A, Q152D, Q152E, Q152H, Q152K, Q152R, Q152Y, D153K, G155F, G155W, G155Y, I156C, I156F, I156M, I156V, P158A, P158G, N159G, N159Q, N159T, N159V, R160A, R 160d, R 160e, R160G, R160H, R160N, R160Q, R160S, R160W, D161I, D161K, D161L, D161M, D161N, D161Q, D161W, A162G, Q163A, Q163C, Q163G, Q163L, Q163M, Q163S, Q163T, P164C, P16 4M, I166L, I166V, A167C, A167E, A167F, A167G, A167K, A167L, A167N, A167Q, A167R, A167T, A167V, A167Y, D168G, D168H, D168L, D168R, D168V, D168W, W169A, W169D, W169E, W169G, W169K, W169Q, W169S, M170F, M170G, M170N, M170Q, M170S, M170V, M170W, K172M, K172P, Q173N, L174A, L174F, L174G, L174T, L174W, Q17517, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617, P17617 L177G, N179H, N179R, N179Y, H180A, H180G, D181H, D181I, D181L, D181R, D181W, S182K, S182L, S182P and / or S182R, where, in the replacement mutation designations, the numbers refer to position positions amino acids of SEQ ID NO: 31. The increase in substrate specificity and / or substrate activity with respect to C12 substrates can be measured in vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que tiene una especificidad por el sustrato, y/o una actividad del sustrato, aumentada (por ejemplo, velocidad catalitica), con respecto a sustratos de C14 (es decir, sustratos, cuyas cadenas de carbono tienen una longitud de 14 carbonos), y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a los restos 5-20, 35-58, 65-80, 83-90, 110-130, 140-145, 155-160, 165-180 de la SEC ID NO: 31. El aumento de la especificidad por el sustrato y/o de la actividad del sustrato con respecto a sustratos de C14 puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which has a specificity for the substrate, and / or an activity of the substrate, increased (for example, catalytic velocity), with respect to C14 substrates ( that is, substrates, whose carbon chains have a length of 14 carbons), and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated in one or more amino acid positions corresponding to residues 5-20, 35-58, 65-80, 83-90, 110-130, 140-145, 155-160, 165- 180 of SEQ ID NO: 31. The increase in substrate specificity and / or substrate activity with respect to C14 substrates can be measured in vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que tiene una especificidad por el sustrato, y/o una actividad del sustrato, aumentada (por ejemplo, velocidad catalitica), con respecto a sustratos de C14 y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a uno o mas restos de la SEC ID NO: 31 seleccionados de 1,4, 5, 7, 8, 9, 11, 12, 13, 14, 15, 16, 17, 20, 21, 22, 23, 25, 26, 28, 29, 33, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 56, 57, 58, 66, 68, 69, 70, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 89, 91, 92, 93, 95, 96, 97, 98, 99, 100, 101, 102, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 131, 133, 134, 136, 137, 138, 139, 140, 141, 142, 143, 147, 148, 151, 152, 153, 155, 156, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 173, 174, 175, 176, 178, 179, 180, 181 y/o 182. El aumento de la especificidad por el sustrato y/o de la actividad del sustrato con respecto a sustratos de C14 puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which has a specificity for the substrate, and / or an activity of the substrate, increased (eg, catalytic velocity), with respect to C14 substrates and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated in one or more amino acid positions corresponding to one or more residues of SEQ ID NO: 31 selected from 1.4, 5, 7, 8, 9, 11, 12, 13, 14 , 15, 16, 17, 20, 21, 22, 23, 25, 26, 28, 29, 33, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 , 49, 50, 51, 52, 53, 54, 56, 57, 58, 66, 68, 69, 70, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83 , 84, 85, 86, 87, 89, 91, 92, 93, 95, 96, 97, 98, 99, 100, 101, 102, 105, 106, 107, 108, 109, 110, 111, 112, 113 , 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 131, 133, 134, 136, 137, 138, 139, 140, 141, 142, 143, 147, 148, 151, 152, 153, 155, 156, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 173, 174, 175, 176, 178, 179, 180, 181 and / or 182. The increase The specificity of the substrate and / or the activity of the substrate with respect to C14 substrates can be measured in vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que tiene una especificidad por el sustrato, y/o una actividad del sustrato, aumentada (por ejemplo, velocidad catalitica), con respecto a sustratos de C14 y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta con una o mas sustituciones seleccionadas de A1S, L4S, L4Y, L5H, L5Y, L7C, L7M, L7N, L7S, L7T, L7Y, G8S, D9N, D9T, L11C, L11I, L11M, L11Q, L11V, S12I, S12L, S12M, S12T, S12V, A13H, A13I, A13L, A13T, A13V, G14F, G14I, G14R, G14T, G14V, Y15A, Y15C, Y15D, Y15E, Y15G, Y15I, Y15L, Y15M, Y15N, Y15Q, Y15R, Y15S, Y15V, R16G, R16N, R16P, R16W, M17C, M17D, M17G, M17K, M17N, M17P, M17R, M17S, M17T, S20A, S20D, S20G, S20L, S20T, S20W, A21G, A22L, A22N, A22Y, W23Y, A25E, A25N, A25Q, A25V, L26C, L26F, L26H, L26Q, L26V, L26Y, N28K, N28P, D29V, S33F, S36H, V37H, V37Q, V38F, N39F, N39M, N39Q, N39V, N39W, N39Y, A40G, A40P, A40T, A40V, S41P, S41T, I42A, I42D, I42E, I42G, I42L, I42M, I42P, I42S, I42T, I42W, I42Y, S43A, S43D, S43E, S43F, S43G, S43H, S43L, S43M, S43N, S43T, S43V, S43W, G44A, G44C, G44E, G44F, G44H, G44K, G44L, G44M, G44N, G44Q, G44R, G44S, G44W, G44Y, D45A, D45C, D45E, D45F, D45G, D45H, D45M, D45P, D45Q, D45S, D45T, D45V, D45W, T46A, T46C, T46D, T46G, T46K, T46N, T46S, T46W, S47E, S47P, S47Q, S47W, S47Y, Q48C, Q48F, Q48I, Q48M, Q48V, Q48W, Q48Y, Q49A, Q49C, Q49D, Q49E, Q49G, Q49H, Q49I, Q49K, Q49L, Q49M, Q49P, Q49R, Q49S, Q49V, Q49W, Q49Y, G50A, G50C, G50E, G50F, G50I, G50K, G50L, G50M, G50N, G50P, G50Q, G50R, G50S, G50T, G50W, G50Y, L51A, L51C, L51D, L51S, L51V, A52H, A52I, A52L, A52M, A52P, A52R, A52V, A52W, A52Y, R53A, R53C, R53D, R53E, R53F, R53G, R53I, R53K, R53L, R53N, R53S, R53T, R53V, R53W, R53Y, L54W, L54Y, A56R, A56W, A56Y, L57F, L58F, L58I, L58Y, V66I, V68L, E69A, E69C, E69D, E69F, E69G, E69H, E69K, E69L, E69M, E69N, E69Q, E69S, E69V, E69Y, L70A, L70C, L70E, L70F, L70H, L70Q, L70S, L70T, L70V, L70W, G72A, G72C, G72P, G72S, N73A, N73C, N73G, N73H, N73I, N73L, N73P, N73R, N73S, N73T, N73V, N73W, D74E, D74G, G75A, G75C, G75D, G75E, G75F, G75I, G75K, G75L, G75M, G75N, G75P, G75T, G75W, G75Y, L76A, L76C, L76D, L76E, L76F, L76G, L76I, L76K, L76M, L76N, L76P, L76Q, L76R, L76T, L76V, L76W, R77A, R77C, R77D, R77E, R77F, R77G, R77H, R77K, R77L, R77N, R77Q, R77S, R77V, R77W, G78P, F79M, F79P, F79V, Q80A, Q80G, Q80L, Q80M, Q80S, Q80W, Q80Y, P81A, P81E, P81K, P81L, P81M, P81W, P81Y, Q82F, Q82I, Q82N, Q82P, Q82V, Q82W, Q82Y, Q83A, T84S, E85D, Q86A, Q86T, Q86V, Q86W, T87A, T87C, T87E, T87F, T87G, T87H, T87L, T87M, T87S, T87V, T87W, R89H, R89T, R89V, R89W, I91L, I91V, I91Y, L92V, Q93A, Q93G, Q93H, Q93I, Q93P, Q93Y, V95L, V95M, V95T, V95W, K96A, K96L, K96P, K96Y, A97W, A98K, A98L, A98W, N99G, N99L, N99P, N99Q, N99R, N99Y, A100G, A100H, A100I, A100K, A100L, A100M, A100R, A100T, A100V, A100Y, E101G, E101L, E101M, E101S, E101T, E101V, P102S, M105A, M105C, M105E, M105G, M105I, M105L, MI05V, Q106A, Q106C, Q106D, Q106G, Q106H, Q106K, Q106L, Q106M, Q106R, Q106S, Q106T, Q106V, Q106W, Q106Y, I107C, I107E, I107G, I107L, I107M, I107Q, I107V, R108A, R108C, R108D, R108F, R108I, R108L, R108S, R108V, R108W, R108Y, L109C, L109M, L109Q, L109T, L109V, L109Y, P110A, P110E, P110H, P110N, P110R, P110V, A111C, A111L, A111Q, A111R, A111V, A111W, N112A, N112F, N112G, N112I, N112L, N112P, N112V, N112W, N112Y, Y113A, Y113D, Y113G, Y113I, Y113M, Y113W, G114F, G114K, G114L, G114M, G114W, G114Y, R115A, R115C, R115E, R115G, R115I, R115N, R115P, R115Q, R115S, R115V, R115W, R115Y, R116C, R116H, R116T, R116V, R116W, Y117C, Y117H, Y117I, Y117L, Y117M, Y117N, YI17S, Y117W, N118A, N118C, N118E, N118G, N118H, N118I, N118L, N118M, N118P, N118Q, N118T, N118V, N118W, E119C, E119D, E119F, E119K, E119M, E119P, E119R, E119T, E119W, E119Y, A120D, A120G, A120I, A120L, A120T, A120W, F121A, F121C, F121D, F121E, F121K, F121L, F121M, F121P, F121Q, F121R, F121S, F121V, F121Y, S122A, S122C, S122D, S122E, S122F, S122G, S122I, S122L, S122M, S122P, S122V, S122W, S122Y, A123C, A123E, A123F, A123H, A123L, A123T, A123V, A123W, A123Y, I124A, I124C, I124G, I124L, I124Y, Y125C, Y125F, Y125G, Y125I, Y125L, Y125P, Y125Q, Y125R, Y125S, Y125T, Y125V, P126C, P126H, P126Y, E131I, E131L, D133K, D133Y, V134S, L136C, L136M, L136Q, L136S, L137P, P138E, P138R, P138T, F139M, F140M, M141A, M141C, M141L, M141P, E142A, E142C, E142L, E142M, E142N, E142P, E142Q, E142S, E142Y, E143I, E143P, K147R, P148W, M151I, M151Q, M151V, Q152A, Q152K, Q152S, D153I, D153K, D153M, D153W, G155F, G155H, G155W, G155Y, I156C, I156F, I156M, I156Q, I156R, I156S, I156V, P158A, P158G, P158S, N159G, N159T, R160A, R160G, R160H, R160N, R160W, D161G, D161I, D161K, D161L, D161M, D161N, D161Q, D161R, D161S, D161V, D161W, A162G, Q163G, Q163L, Q163M, Q163S, P164A, P164C, P164K, P164L, P164M, P164N, P164R, P164T, P164W, F165G, F165H, F165S, F165W, F165Y, I166L, I166V, A167T, D168A, D168G, D168H, D168P, D168R, D168T, W169A, W169E, W169K, W169M, W169Q, W169R, W169S, W169T, W169V, M170A, M170F, M170V, A171I, Q173N, Q173W, Q173Y, L174Q, L174W, Q175I, Q175Y, P176H, P176K, P176L, P176R, P176W, P176Y, V178A, V178T, V178W, N179H, N179R, N179T, N179V, N179Y, H180G, H180R, H180S, H180W, D181A, D181H, D181I, D181L, D181Q, D181R, D181S, D181W, S182A, S182E, S182G, S182I, S182K, S182L, S182P, S182Q, S182R y/o S182T, en donde, en las designaciones de mutacion por sustitucion, los numeros se refieren a posiciones de aminoacidos de la SEC ID NO: 31. El aumento de la especificidad por el sustrato y/o de la actividad del sustrato con respecto a sustratos de C14 puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which has a specificity for the substrate, and / or an activity of the substrate, increased (eg, catalytic velocity), with respect to C14 substrates and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated with one or more substitutions selected from A1S, L4S, L4Y, L5H, L5Y, L7C, L7M, L7N, L7S, L7T, L7Y, G8S, D9N, D9T, L11C, L11I, L11M, L11Q, L11V, S12I, S12L, S12M, S12T, S12V, A13H, A13I, A13L, A13T, A13V, G14F, G14I, G14R, G14T, G14V, Y15A, Y15C, Y15D, Y15E, Y15G, Y15I, Y15L, Y15L, Y15L, Y15L Y15N, Y15Q, Y15R, Y15S, Y15V, R16G, R16N, R16P, R16W, M17C, M17D, M17G, M17K, M17N, M17P, M17R, M17S, M17T, S20A, S20D, S20G, S20L, S20T, S20W, A20W, A20W A22L, A22N, A22Y, W23Y, A25E, A25N, A25Q, A25V, L26C, L26F, L26H, L26Q, L26V, L26Y, N28K, N28P, D29V, S33F, S36H, V37H, V37Q, V38F, N39F, N39M, N39M, N39M, N39M, N39M, N39M N39V, N39W, N39Y, A40G, A40P, A40T, A40V, S41P, S41T, I42A, I42D, I42E, I42G, I42L, I42M, I42P, I42S, I42T, I42W, I42Y, S43A, S43D, S43E, S43F, S43F, S43F, S43F S43H, S43L, S43M, S43N, S43T, S43V, S43W, G44A, G44C, G44E, G44F, G44H, G44K, G44L, G44M, G44N, G44Q, G44R, G44S, G44W, G44Y, D45A, D45C, D45E, D45E D45G, D45H, D45M, D45P, D45Q, D45S, D45T, D45V, D45W, T46A, T46C, T46 D, T46G, T46K, T46N, T46S, T46W, S47E, S47P, S47Q, S47W, S47Y, Q48C, Q48F, Q48I, Q48M, Q48V, Q48W, Q48Y, Q49A, Q49C, Q49D, Q49E, Q49G, Q49H, Q49I, Q49I Q49K, Q49L, Q49M, Q49P, Q49R, Q49S, Q49V, Q49W, Q49Y, G50A, G50C, G50E, G50F, G50I, G50K, G50L, G50M, G50N, G50P, G50Q, G50R, G50S, G50T, G50W, G50W, G50W L51A, L51C, L51D, L51S, L51V, A52H, A52I, A52L, A52M, A52P, A52R, A52V, A52W, A52Y, R53A, R53C, R53D, R53E, R53F, R53G, R53I, R53K, R53L, R53N, R53N, R53N R53T, R53V, R53W, R53Y, L54W, L54Y, A56R, A56W, A56Y, L57F, L58F, L58I, L58Y, V66I, V68L, E69A, E69C, E69D, E69F, E69G, E69H, E69K, E69L, E69M, E69M, E69M, E69M E69Q, E69S, E69V, E69Y, L70A, L70C, L70E, L70F, L70H, L70Q, L70S, L70T, L70V, L70W, G72A, G72C, G72P, G72S, N73A, N73C, N73G, N73H, N73I, N73L, N73L, N73L, N73L, N73L N73R, N73S, N73T, N73V, N73W, D74E, D74G, G75A, G75C, G75D, G75E, G75F, G75I, G75K, G75L, G75M, G75N, G75P, G75T, G75W, G75Y, L76A, L76C, L76D, L76D, L76D, L76D, L76D, L76D L76F, L76G, L76I, L76K, L76M, L76N, L76P, L76Q, L76R, L76T, L76V, L76W, R77A, R77C, R77D, R77E, R77F, R 77G, R77H, R77K, R77L, R77N, R77Q, R77S, R77V, R77W, G78P, F79M, F79P, F79V, Q80A, Q80G, Q80L, Q80M, Q80S, Q80W, Q80Y, P81A, P81E, P81K, P81L, P81M P81W, P81Y, Q82F, Q82I, Q82N, Q82P, Q82V, Q82W, Q82Y, Q83A, T84S, E85D, Q86A, Q86T, Q86V, Q86W, T87A, T87C, T87E, T87F, T87G, T87H, T87L, T87M, T87L, T87M, T87M T87V, T87W, R89H, R89T, R89V, R89W, I91L, I91V, I91Y, L92V, Q93A, Q93G, Q93H, Q93I, Q93P, Q93Y, V95L, V95M, V95T, V95W, K96A, K96L, K96P, K96Y, A97 A98K, A98L, A98W, N99G, N99L, N99P, N99Q, N99R, N99Y, A100G, A100H, A100I, A100K, A100L, A100M, A100R, A100T, A100V, A100Y, E101G, E101L, E101M, E101S, E101S, E101S, E10110 P102S, M105A, M105C, M105E, M105G, M105I, M105L, MI05V, Q106A, Q106C, Q106D, Q106G, Q106H, Q106K, Q106L, Q106M, Q106R, Q106S, Q106T, I106, Q106, I107 I107L, I107M, I107Q, I107V, R108A, R108C, R108D, R108F, R108I, R108L, R108S, R108V, R108W, R108Y, L109C, L109M, L109Q, L109T, L109V, L109Y, P110A, P110E, P110H, P110N, P110R, P110V, A111C, A111L, A111Q, A111R, A111V, A111W, N112A, N112F, N112G, N112I, N112L, N112P, N112V, N112W, N112Y, Y113A, Y113D, Y113G, Y113I, Y113M, Y113W, G114F, G114K, R114, R114, R115, G114, R115, G114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R11, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R114, R115 R115E, R115G, R115I, R115N, R115P, R115Q, R115S, R115V, R115W, R115Y, R116C, R116H, R116T, R116V, R116W, Y117C, Y117H, Y117I, Y117L, Y117M, Y117N, YI17S, Y117W, N118A, N118C, N118E, N118G, N118H, N118I, N118L, N118M, N118P, N118Q, N118T, N118V, N118W, E119C, E119D, E119F, E119K, E119M, E119P, E119R, E119T, E11920, A120, A1201 A120T, A120W, F121A, F121C, F121D, F121E, F121K, F121L, F121M, F121P, F121Q, F121R, F121S, F121V, F121Y, S122A, S122C, S122D, S122E, S122F, S122F, S122F, S122F, S122M, S122M, S122M, S122M, S122M, S122M, S122M, S122M, S122M, S122M, S122M, S122M, S122M, S122M, S122M2 S122V, S122W, S122Y, A123C, A123E, A123F, A123H, A123L, A123T, A123V, A123W, A123Y, I124A, I124C, I124G, I124L, I124Y, Y125C, Y125F, Y125G, Y12512, Y12512, Y12512 Y125S, Y125T, Y125V, P126C, P126H, P126Y, E131I, E131L, D133K, D133Y, V134S, L136C, L136M, L136Q, L136S, L137P, P138E, P138R, P138T, F139M, F140M, M141A, M141C, M141L, M141P, E142A, E142C, E142L, E142M, E142N, E142P, E142Q, E142S, E142Y, E143I, E143P, K147R, Q158, Q158, Q158, Q15, Q15, Q15, Q15, Q15, Q15, Q15 Q152S, D153I, D153K, D153M, D153W, G155F, G155H, G155W, G155Y, I156C, I156F, I156M, I156Q, I156R, I156S, I156V, P158A, P158G, P15816, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R160, R15 R160W, D161G, D161I, D161K, D161L, D161M, D161N, D161Q, D161R, D161S, D161V, D161W, A162G, Q163G, Q163L, Q163M, Q163S, P164A, P164C, P164C, P164C, P164M, P164M, P164M, P164M, P164M P164W, F165G, F165H, F165S, F165W, F165Y, I166L, I166V, A167T, D168A, D168G, D168H, D168P, D168R, D168T, W169A, W169E, W169K, W169M, W169Q, W169R, W169S, W169T, W169V, M170A, M170F, M170V, A171I, Q173N, Q173W, Q173Y, L174Q, L174W, Q175I, Q175Y, P176H, P176K, P176L, P176R, P176W, P176Y, V178A, V178T, V178W, N179, N179, N179, N179, N179 H180R, H180S, H180W, D181A, D181H, D181I, D181L, D181Q, D181R, D181S, D181W, S182A, S182E, S182G, S182I, S182K, S182L, S182P , S182Q, S182R and / or S182T, where, in the substitution mutation designations, the numbers refer to amino acid positions of SEQ ID NO: 31. The increase in specificity for the substrate and / or activity of the substrate with respect to C14 substrates can be measured in vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que tiene preferencia por sustratos de ester (por ejemplo, acil-PNP) sobre sustratos de tioester (por ejemplo, acil-CoA), y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a restos seleccionados de 95, 96, 97, 98, 99, 100, 101, 102, 104, 105, 106, 107, 108, 109 y/o 110 de la SEC ID NO: 31. La preferencia por sustratos de ester sobre sustratos de tioester puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which has preference for ester substrates (e.g., acyl-PNP) over thioester substrates (e.g., acyl-CoA), and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated in one or more amino acid positions corresponding to residues selected from 95, 96, 97, 98, 99, 100, 101, 102, 104, 105, 106, 107, 108, 109 and / or 110 of SEQ ID NO: 31. The preference for ester substrates over thioester substrates can be measured in vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que tiene preferencia por sustratos de ester (por ejemplo, acil-PNP) sobre sustratos de tioester (por ejemplo, acil-CoA), y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta con una o mas sustituciones seleccionadas de V95L, V95M, V95T, K96A, K96L, K96W, K96Y, A97F, A97K, A97S, A97T, A97W, A98E, A98F, A98K, A98L, A98Q, A98W, N99Y, A100K, A100V, E101L, P102S, L104C, M105F, Q106A, Q106C, Q106T, Q106Y, I107A, I107C, I107G, I107L, I107M, I107Q, I107V, R108A, R108C, R108D, R108F, R108I, R108L, R108S, R108V, R108W, R108Y, L109M, L109V, P110A, P110F, P110H, P110N, P110V y/o P110W, en donde, en las designaciones de mutacion por sustitucion, los numeros se refieren a posiciones de aminoacidos de la SEC ID NO: 31. La preferencia por sustratos de ester sobre sustratos de tioester puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which has preference for ester substrates (e.g., acyl-PNP) over thioester substrates (e.g., acyl-CoA), and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated with one or more substitutions selected from V95L, V95M, V95T, K96A, K96L, K96W, K96Y, A97F, A97K, A97S, A97T, A97W, A98E, A98F, A98K, A98L, A98Q, A98W, N99Y, A100K, A100V, E101L, P102S, L104C, M105F, Q106A, Q106C, Q106T, Q106Y, I107A, I107C, I107G, I107L, I107M, I107Q, I107V, R10810, R810, R810, R810, R810 R108S, R108V, R108W, R108Y, L109M, L109V, P110A, P110F, P110H, P110N, P110V and / or P110W, where, in the substitution mutation designations, the numbers refer to amino acid positions of SEQ ID NO : 31. The preference for ester substrates over thioester substrates can be measured in vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que tiene preferencia por sustratos de tioester (por ejemplo, acil-CoA) sobre sustratos de ester (por ejemplo, acil-PNP), y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a restos seleccionados de 95, 96, 97, 101, 102, 103, 104, 105, 107, 109 y/o 110 de la SEC ID NO: 31. La preferencia por los sustratos de tioester sobre los sustratos de ester puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which has a preference for thioester substrates (e.g., acyl-CoA) over ester substrates (e.g., acyl-PNP), and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated at one or more amino acid positions corresponding to moieties selected from 95, 96, 97, 101, 102, 103, 104, 105, 107, 109 and / or 110 of SEQ ID NO: 31. The preference for thioester substrates over ester substrates can be measured in vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que tiene preferencia por sustratos de tioester (por ejemplo, acil-CoA) sobre sustratos de ester (por ejemplo, acil-PNP), y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta con una o mas sustituciones seleccionadas de V95E, V95I, V95W, V95Y, K96P, A97E, A97M, E101P, P102D, P102K, P102Y, L103E, L103K, L103N, L104A, L104D, L104E, L104N, L104Q, L104W, L104Y, M105W, I107E, I107K, I107P, L109A, L109C, L109D, L109E, L109G, L109K, L109N, L109P, L109Q, L109S, L109T, L109Y y/o P110R, en donde, en las designaciones de mutacion por sustitucion, los numeros se refieren a posiciones de aminoacidos de la SEC ID NO: 31. La preferencia por los sustratos de tioester sobre los sustratos de ester puede medirse in vitro y/o in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which has a preference for thioester substrates (e.g., acyl-CoA) over ester substrates (e.g., acyl-PNP), and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated with one or more substitutions selected from V95E, V95I, V95W, V95Y, K96P, A97E, A97M, E101P, P102D, P102K, P102Y, L103E, L103K, L103N, L104A, L104D, L104D, L104D, L104D, L104D L104N, L104Q, L104W, L104Y, M105W, I107E, I107K, I107P, L109A, L109C, L109D, L109E, L109G, L109K, L109N, L109P, L109Q, L109S, L109T or L109T, L109T, L109T, L109T, L109T, L109T, L109T, L109T or L109T By substitution mutation, the numbers refer to amino acid positions of SEQ ID NO: 31. The preference for thioester substrates over ester substrates can be measured in vitro and / or in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que es capaz de producir un aumento del rendimiento proporcional o porcentual de esteres grasos sobre otros productos de esteres no grasos (por ejemplo, acidos grasos libres y/o derivados de acidos grasos distintos de los esteres grasos), y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEQ ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a los restos de la SEC ID NO: 31 seleccionados de 1-14, 22-29, 33-58, 65-100, 103-109, 114-117, 119-121, 127-136, 139-144, 150-151, 155-170 y / o 173-174. El aumento del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, derivados de acidos grasos distintos de esteres grasos) puede observarse o determinarse in vitro y/o in vivo. Preferentemente, el aumento del rendimiento proporcional o porcentual de esteres grasos sobre otros productos se determina in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which is capable of producing an increase in the proportional or percentage yield of fatty esters over other non-fatty ester products (for example, free fatty acids and / or derived from fatty acids other than fatty esters), and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated in one or more amino acid positions corresponding to the residues of SEQ ID NO: 31 selected from 1-14, 22-29, 33-58, 65-100, 103-109, 114 -117, 119-121, 127-136, 139-144, 150-151, 155-170 and / or 173-174. The increase in the proportional or percentage yield of fatty esters on other products (for example, derivatives of fatty acids other than fatty esters) can be observed or determined in vitro and / or in vivo. Preferably, the increase in the proportional or percentage yield of fatty esters on other products is determined in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que es capaz de producir un aumento del rendimiento proporcional o porcentual de esteres grasos sobre otros productos de esteres grasos (por ejemplo, acidos grasos libres y/o derivados de acidos grasos distintos de esteres grasos), y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEQ ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a los restos de la SEC ID NO: 31 seleccionados de 1,2, 4, 5, 6 , 7, 8, 12, 13, 14, 22, 23, 24, 25, 26, 28, 29, 33, 34, 35, 36, 37, 38, 39, 40, 41, 44, 45, 46, 47, 49, 50, 53, 58, 65, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 79, 81, 84, 86, 87, 88, 89, 90, 91, 92, 93, 95, 96, 99, 100, 103, 104, 105, 106, 107, 108, 109, 114, 115, 117, 119, 120, 121, 127, 128, 129, 131, 132, 134, 135, 136, 139, 141, 142, 143, 144, 150, 151, 155, 156, 158, 159, 160, 161, 162, 163, 164, 165, 166, 169, 170, 173 y / o 174. El aumento del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, derivados de acidos grasos distintos de esteres grasos) puede observarse o determinarse in vitro y/o in vivo. Preferentemente, el aumento del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, derivados de acidos grasos distintos de esteres grasos) se determina in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which is capable of producing an increase in the proportional or percentage yield of fatty esters over other fatty ester products (for example, free fatty acids and / or derivatives). of fatty acids other than fatty esters), and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated in one or more amino acid positions corresponding to the residues of SEQ ID NO: 31 selected from 1,2, 4, 5, 6, 7, 8, 12, 13, 14, 22 , 23, 24, 25, 26, 28, 29, 33, 34, 35, 36, 37, 38, 39, 40, 41, 44, 45, 46, 47, 49, 50, 53, 58, 65, 67 , 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 79, 81, 84, 86, 87, 88, 89, 90, 91, 92, 93, 95, 96, 99, 100 , 103, 104, 105, 106, 107, 108, 109, 114, 115, 117, 119, 120, 121, 127, 128, 129, 131, 132, 134, 135, 136, 139, 141, 142, 143 , 144, 150, 151, 155, 156, 158, 159, 160, 161, 162, 163, 164, 165, 166, 169, 170, 173 and / or 174. The increase in the proportional or percentage yield of fatty esters on Other products (for example, derivatives of fatty acids other than fatty esters) can be observed or determined in vitro and / or in vivo. Preferably, the increase in the proportional or percentage yield of fatty esters on other products (for example, derivatives of fatty acids other than fatty esters) is determined in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que es capaz de producir un aumento del rendimiento proporcional o porcentual de esteres grasos sobre otros productos de esteres grasos (por ejemplo, acidos grasos libres y/o derivados de acidos grasos distintos de esteres grasos), y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEQ ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta con una o mas sustituciones seleccionadas de A1R, D2H, D2R, L4G, L4M, L5Q, I6A, I6L, L7E, G8A, S12N, A13I, A13L, A13S, A13T, A13W, A13Y, G14K, G14R, G14S, G14T, A22D, A22E, A22H, A22Y, W23Y, P24C, P24G, P24T, A25P, L26C, L26D, L26E, L26G, L26N, N28A, N28M, D29V, S33G, S33M, K34A, K34H, K34M, T35G, T35M, S36A, V37A, V37G, V37H, V37S, V38D, V38G, V38P, N39E, N39Q, N39R, A40M, A40P, S41T, G44F, G44Y, D45P, D45Q, T46W, S47F, Q49I, G50A, G50K, G50M, G50S, R53S, L58D, L58M, L58R, W65L, L67G, V68G, V68M, V68N, E69P, E69Q, L70A, L70E, L70H, G71C, G72A, N73C, N73G, N73L, N73R, N73T, N73V, D74C, D74S, D74W, G75A, G75K, G75L, G75M, L76A, L76F, L76G, L76I, L76M, L76N, L76T, L76W, R77G, F79A, F79M, F79P, P81E, P81W, T84F, T84H, T84Y, Q86P, Q86W, T87M, T87S, T87W, L88C, L88F, L88G, L88H, L88Y, R89G, Q90P, Q90W, I91M, I91S, L92C, L92G, Q93F, Q93P, V95A, V95D, V95E, V95L, V95M, K96P, N99L, N99M, N99S, A100D, A100K, A100L, A100M, A100V, A100Y, L103A, L104A, L104C, L104P, L104Q, L104W, M105A, Q106A, Q106C, Q106T, Q106W, I107C, I107M, R108E, L109F, L109M, G114F, R115W, Y117P, E119D, E119P, A120P, F121A, F121C, F121W, K127P, L128F, A129L, A129Y, E131A, F132P, V134P, P135A, L136A, F139M, M141A, M141P, E142A, E143P, V144A, W150D, W150E, M151S, G155V, I156K, I156M, P158A, P158G, P158Q, P158S, N159E, N159I, R160H, R160I, R160K, D161G, A162T, A162Y, Q163A, Q163C, Q163E, Q163G, Q163I, Q163M, Q163S, Q163T, Q163V, P164C, F165D, F165S, I166A, I166L, W169M, M170E, M170G, M170N, M170S, Q173P y/o L174A, en donde, en las designaciones de mutacion por sustitucion, los numeros se refieren a posiciones de aminoacidos de la SEC ID NO: 31. El aumento del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, derivados de acidos grasos distintos de esteres grasos) puede observarse o determinarse in vitro y/o in vivo. Preferentemente, el aumento del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, derivados de acidos grasos distintos de esteres grasos) se determina in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which is capable of producing an increase in the proportional or percentage yield of fatty esters over other fatty ester products (for example, free fatty acids and / or derivatives). of fatty acids other than fatty esters), and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated with one or more substitutions selected from A1R, D2H, D2R, L4G, L4M, L5Q, I6A, I6L, L7E, G8A, S12N, A13I, A13L, A13S, A13T, A13W, A13Y, G14K, G14R, G14S, G14T, A22D, A22E, A22H, A22Y, W23Y, P24C, P24G, P24T, A25P, L26C, L26D, L26E, L26G, L26N, N28A, N28M, D29V, S33G, S33M, K34A, K34A, K34A, K34A K34M, T35G, T35M, S36A, V37A, V37G, V37H, V37S, V38D, V38G, V38P, N39E, N39Q, N39R, A40M, A40P, S41T, G44F, G44Y, D45P, D45Q, T46W, S47FA, Q49I, G49I G50K, G50M, G50S, R53S, L58D, L58M, L58R, W65L, L67G, V68G, V68M, V68N, E69P, E69Q, L70A, L70E, L70H, G71C, G72A, N73C, N73G, N73L, N73R, N73T, N73V, D74C, D74S, D74W, G75A, G75K, G75K G75M, L76A, L76F, L76G, L76I, L76M, L76N, L76T, L76W, R77G, F79A, F79M, F79P, P81E, P81W, T84F, T84H, T84Y, Q86P, Q86W, T87M, T87S, T87W, L88C, L88C, L88C, L88C L88G, L88H, L88Y, R89G, Q90P, Q90W, I91M, I91S, L92C, L92G, Q93F, Q93P, V95A, V95D, V95E, V95L, V95M, K96P, N99L, N99M, N99S, A100D, A100K, A100L, A100L, A100L, A100L, A100L, A100L A100V, A100Y, L103A, L104A, L104C, L104P, L104Q, L104W, M105A, Q106A, Q106C, Q106T, Q106W, I107C, I107M, R108E, L109F, L109M, G114F, R115, F115, F115, F115, F115, F115, F115, F115 F121C, F121W, K127P, L128F, A129L, A129Y, E131A, F132P, V134P, P135A, L136A, F139M, M141A, M141P, E142A, E143P, V144A, W150D, W150E, M151S, G155V, I156K, I156M, P158A, P158G, P158Q, P158S, N159E, N159I, R160H, R160I, R160K, D161G, A162T, A162Y, Q163A, Q163C, Q163E, Q163G, Q163I, Q163M, Q163S, Q163T, Q163V, P164C, F165D, F165S, I166A, I166L, W169M, M170E, M170G, M170N, M170S, Q173P and / or L174A, where, in the substitution mutation designations, the numbers refer to amino acid positions of SEQ ID NO: 31. The increase in the proportional or percentage yield of fatty esters on other products (by example, derivatives of fatty acids other than fatty esters) can be observed or determined in vitro and / or in vivo. Preferably, the increase in the proportional or percentage yield of fatty esters on other products (for example, derivatives of fatty acids other than fatty esters) is determined in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que es capaz de producir una disminucion del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, acidos grasos libres y/o derivados de acidos grasos distintos de esteres grasos) cuando la produccion de esteres grasos no es deseable, y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEQ ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a los restos de la SEC ID NO: 31 seleccionados de 3, 5, 15-18, 27-42, 46, 57-68, 77­ 78, 95-106, 121-123, 152-154, 167 y/o 175-182. La disminucion del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, derivados de acidos grasos distintos de esteres grasos) puede observarse o determinarse in vitro y/o in vivo. Preferentemente, la disminucion del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, derivados de acidos grasos distintos de esteres grasos) se determina in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which is capable of producing a decrease in the proportional or percentage yield of fatty esters on other products (for example, free fatty acids and / or fatty acid derivatives other than fatty esters) when the production of fatty esters is undesirable, and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated in one or more amino acid positions corresponding to the residues of SEQ ID NO: 31 selected from 3, 5, 15-18, 27-42, 46, 57-68, 77 78, 95-106, 121-123, 152-154, 167 and / or 175-182. The decrease in the proportional or percentage yield of fatty esters on other products (for example, derivatives of fatty acids other than fatty esters) can be observed or determined in vitro and / or in vivo. Preferably, the decrease in the proportional or percentage yield of fatty esters on other products (for example, derivatives of fatty acids other than fatty esters) is determined in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que es capaz de producir una disminucion del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, acidos grasos libres y/o derivados de acidos grasos distintos de esteres grasos) cuando la produccion de esteres grasos no es deseable, y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEQ ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a los restos de la SEC ID NO: 31 seleccionados de 3, 5, 15, 16, 18, 27, 28, 33, 34, 35, 36, 37, 38, 40, 42, 46, 57, 59, 60, 62, 65, 68, 77, 78, 95, 96, 97, 98, 99, 100, 102, 103, 105, 106, 121, 123, 152, 153, 154, 167, 175, 176, 178, 179, 180, 181 y/o 182. La disminucion del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, derivados de acidos grasos distintos de esteres grasos) puede observarse o determinarse in vitro y/o in vivo. Preferentemente, la disminucion del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, derivados de acidos grasos distintos de esteres grasos) se determina in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which is capable of producing a decrease in the proportional or percentage yield of fatty esters on other products (for example, free fatty acids and / or fatty acid derivatives other than fatty esters) when the production of fatty esters is undesirable, and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated in one or more amino acid positions corresponding to the residues of SEQ ID NO: 31 selected from 3, 5, 15, 16, 18, 27, 28, 33, 34, 35, 36 , 37, 38, 40, 42, 46, 57, 59, 60, 62, 65, 68, 77, 78, 95, 96, 97, 98, 99, 100, 102, 103, 105, 106, 121, 123 , 152, 153, 154, 167, 175, 176, 178, 179, 180, 181 and / or 182. The decrease in the proportional or percentage yield of fatty esters on other products (for example, derivatives of fatty acids other than fatty esters ) can be observed or determined in vitro and / or in vivo. Preferably, the decrease in the proportional or percentage yield of fatty esters on other products (for example, derivatives of fatty acids other than fatty esters) is determined in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que es capaz de producir una disminucion del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, acidos grasos libres y/o derivados de acidos grasos distintos de esteres grasos) cuando la produccion de esteres grasos no es deseable, y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEQ ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta con una o mas sustituciones seleccionadas de T3E, T3G, T3K, T3L, L5C, L5G, Y15A, Y15L, Y15Q, Y15R, Y15V, R16D, R16E, R16G, R16I, R16V, S18E, L27V, N28G, N28I, S33I, S33R, K34R, T35F, T35K, T35L, T35Q, T35V, S36F, S36I, S36L, S36W, V37L, V38E, V38F, V38K, V38L, A40D, A40G, I42T, T46L, L57A, L57F, L57G, L57H, L57K, L57N, L57P, L57R, L57S, L57T, L57V, L57W, L57Y, K59V, Q60E, Q60P, Q62G, W65V, V68L, R77L, G78M, V95F, V95N, K96C, K96L, K96N, K96Q, K96R, K96Y, A97E, A97F, A97R, A97W, A98E, N99A, N99D, A100S, P102I, L103Q, L103W, M105L, Q106G, Q106H, Q106K, Q106S, Q106V, F121P, A123E, Q152D, Q152E, Q152F, Q152H, Q152I, Q152K, Q152L, Q152S, Q152T, Q152Y, D153P, D153V, D154E, A167V, Q175L, P176D, V178K, N179H, N179W, H180E, H180L, H180P, H180R, D181C, D181E, S182K, S182L, S182N, S182R, S182T y/o S182V, en donde, en las designaciones de mutacion por sustitucion, los numeros se refieren a posiciones de aminoacidos de la SEC ID NO: 31. La disminucion del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, derivados de acidos grasos distintos de esteres grasos) puede observarse o determinarse in vitro y/o in vivo. Preferentemente, la disminucion del rendimiento proporcional o porcentual de esteres grasos sobre otros productos (por ejemplo, derivados de acidos grasos distintos de esteres grasos) se determina in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which is capable of producing a decrease in the proportional or percentage yield of fatty esters on other products (for example, free fatty acids and / or fatty acid derivatives other than fatty esters) when the production of fatty esters is undesirable, and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated with one or more substitutions selected from T3E, T3G, T3K, T3L, L5C, L5G, Y15A, Y15L, Y15Q, Y15R, Y15V, R16D, R16E, R16G, R16I, R16V, S18E, L27V, N28G, N28I, S33I, S33R, K34R, T35F, T35K, T35L, T35Q, T35V, S36F, S36I, S36L, S36W, V37L, V38E, V38F, V38K, V38L, A40D, A40G, I42T, T46L, L57 L57F, L57G, L57H, L57K, L57N, L57P, L57R, L57S, L57T, L57V, L57W, L57Y, K59V, Q60E, Q60P, Q62G, W65V, V68L, R77L, G78M, V95F, V95N, K96C, K96L, K96L, K96L, K96L K96Q, K96R, K96Y, A97E, A97F, A97R, A97W, A98E, N99A, N99D, A100S, P102I, L103Q, L103W, M105L, Q106G, Q106H, Q106K, Q106S, Q106V, F121P2, A123E, Q123E Q152H, Q152I, Q152K, Q152L, Q152S, Q152T, Q152Y, D153P, D153V, D154E, A167V, Q175L, P176D, V178K, N179H, N179W, H180E, H180L, H180P, H180R, D181C, D181E, S182K, S182L, S182N, S182R, S182T and / or S182V, where, in the substitution mutation designations, the numbers refer to amino acid positions of SEQ ID NO: 31. The decrease The proportional or percentage yield of fatty esters on other products (for example, derivatives of fatty acids other than fatty esters) can be observed or determined in vitro and / or in vivo. Preferably, the decrease in the proportional or percentage yield of fatty esters on other products (for example, derivatives of fatty acids other than fatty esters) is determined in vivo.

En un aspecto, se desvela una tioesterasa mutante (o uno de sus equivalentes naturales), que es capaz de aumentar y/o mejorar la produccion de uno o mas derivados de acidos grasos, y que es una variante de una tioesterasa precursora que comprende una secuencia analoga a la SEC ID NO: 31 expuesta en la FIG. 58, en donde la tioesterasa precursora se muta en una o mas posiciones de aminoacidos correspondientes a los restos de la SEC ID NO: 31 seleccionados de 2, 4, 11-22, 25-31, 37-45, 49-58, 63-80, 84- 130, 136-146 y/o 150-174. Un derivado de ac ido g ra so e je m p la r que se p roduce de a cu e rd o con esto es un ac ido g raso libre. El a u m en to y /o la m e jo ra de la p ro du cc ion de de riva dos de ac ido s g ra sos puede m ed irse in vitro y /o in vivo. P re fe re n tem e n te , el au m e n to y /o la m e jo ra de la p ro du cc ion de d e riva d o s de ac ido s g ra sos se m ide in vivo.

In one aspect, a mutant thioesterase (or one of its natural equivalents) is disclosed, which is capable of increasing and / or improving the production of one or more fatty acid derivatives, and which is a variant of a precursor thioesterase comprising a sequence analogous to SEQ ID NO: 31 set forth in FIG. 58, wherein the precursor thioesterase is mutated in one or more amino acid positions corresponding to the residues of SEQ ID NO: 31 selected from 2, 4, 11-22, 25-31, 37-45, 49-58, 63 -80, 84- 130, 136-146 and / or 150-174. A derivative of This is a great idea of how it is a free common acid. The increase in and / or the improvement of the de riva dos de riva dos de ac gone can also be in vitro and / or in vivo. P re fe re n tem in te, the increase and / or the improvement of the de riva dos de riva dos de ac gone s se se ra idem in vivo.

En un aspecto , se de sve la una tio e s te ra sa m u ta n te (o uno de sus eq u iva le n te s na tu ra les), que es cap az de a u m e n ta r y /o m e jo ra r la p ro du cc ion de uno o m as de riva dos de ac ido s g rasos, y que es una va ria n te de una t io e s te ra sa p re cu rso ra que com p ren de una secu en c ia a n a loga de la S E C ID NO: 31 exp ue s ta en la FIG. 58, en do nde la t io e s te ra sa p re cu rso ra se m uta en una o m as p o s ic io ne s de a m in o a c id o s co rre sp o n d ie n te s a los res tos de la S E C ID NO: 31 se le c c io n a d o s de 2, 4, 11, 12, 13, 14, 15, 16, 17, 19, 21, 22, 25, 26, 27, 28, 29, 30, 31, 37, 39, 41, 42, 43, 44, 45, 49, 50, 51, 53, 54, 58, 63, 65, 66, 67, 68, 69, 70, 71 ,73 , 74, 75, 76, 77, 78, 79, 80, 84, 87, 88, 90, 91 ,92 , 93, 94, 95, 96, 97, 98, 100, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 115, 117, 118, 119, 120, 121, 122, 124, 127, 128, 129, 130, 136, 137, 138, 139, 140, 141, 143, 144, 145, 146, 150, 151, 152, 154, 155, 156, 158, 162, 163, 166, 167, 169, 170, 173 y/o 174. Un de riva d o de ac ido g raso e je m p la r que se p roduce de a cu e rd o con esto es un ac ido g ra so libre. El a u m en to y/o la m e jo ra de la p ro du cc ion de un de riva d o de ac ido graso puede m ed irse in vitro y /o in vivo. P re fe re n tem e n te , el a u m en to y /o la m e jo ra de la p ro du cc ion de un de riva do de ac ido g ra so se m ide in vivo.

In one aspect, it is only one uncle is te ra sa mu ta n te (or one of his eq u iva le n te s na tu ra rales), which is the capital of the increase and omega ra la For one or more of the two years of succession, and that is a variant of an uncle, it is the reason for the purchase of a sector in one year. according to SEQ ID NO: 31 set forth in FIG. 58, where the uncle is the one who is killed in one or more positions in the United States or the rest of the SEQ ID NO: 31 were given 2, 4, 11, 12, 13, 14, 15, 16, 17, 19, 21, 22, 25, 26, 27, 28, 29, 30, 31, 37 , 39, 41, 42, 43, 44, 45, 49, 50, 51, 53, 54, 58, 63, 65, 66, 67, 68, 69, 70, 71, 73, 74, 75, 76, 77 , 78, 79, 80, 84, 87, 88, 90, 91, 92, 93, 94, 95, 96, 97, 98, 100, 102, 103, 104, 105, 106, 107, 108, 109, 110 , 111, 112, 113, 115, 117, 118, 119, 120, 121, 122, 124, 127, 128, 129, 130, 136, 137, 138, 139, 140, 141, 143, 144, 145, 146 , 150, 151, 152, 154, 155, 156, 158, 162, 163, 166, 167, 169, 170, 173 and / or 174. One of the largest private areas in the country He argues from this when he is a free agent. The increase in and / or the improvement of a fatty acid source can be in vitro and / or in vivo. P re fe re n tem in te, the increase in and / or the improvement of the protection of a source of income is only in vivo.

En un aspecto , se de sve la una tio e s te ra sa m u ta n te (o uno de sus eq u iva le n te s na tu ra les), que es cap az de a u m e n ta r y /o m e jo ra r la p ro du cc ion de uno o m as de riva dos de ac ido s g rasos, y que es una va ria n te de una t io e s te ra s a p re cu rso ra que co m p re n d e una se cu e n c ia a n a lo g a a la S E C ID NO: 31 e xp u e s ta en la FIG. 58, en dond e la t io e s te ra s a p re cu rso ra se m u ta con una o m as s u s titu c io n e s se le c c io n a d a s de: D 2L, D 2P , D2R, L5G , L11I, S 12N , S 12T , A 13N , G 14C , G 14P , G 14S , G 14T , G 14V , Y 15C , Y 15I, Y 15V , R16T, M 17D , M 17E, M 17N , M 17R , M 17S, M 17V , A 19C , A 21G , A 22 L , A 22R , A 22 T , A 25P , L26D , L26G , L26W , L27C , L27F, L27W , L27Y , N28I, N28P , D29P, K30P , W 31D , W 31G , W 31N , W 31P , W 31R , W 31S , W 31T , V 37Y , N 39P , S 41C , I42D, I42G , S43E , G 44K , G 44R , G 44W , D 45G , Q 49E , G 50A , G 50K , G 50M , G 50Q , L51D , L51T , R 53A , R 53G , R 53L, R 53N , R53S, R 53V , L54E, L54F , L54G , L54N , L54S , L54W , L58R , P63G , P 63M , P63N , P 63T, P 63W , W 65E , W 65G , V 66G , V 66S , L67T , V 68S , E 69F , E 69V , L70C , L70F , L70Q , L70S , L70T , L70V , G 71A , N 73G , N 73L, D 74A , D 74C , G 75A , G 75C , G 75F , G 75R , G 75W , L76I, R 77A , R 77C , R 77D , R77F, R 77G , R 77H , R 77K , R 77L, R 77N , R 77Q , R 77S , R 77W , G 78D , G 78E , F79K , Q 80G , T84H , T 84N , T84Q , T87A , T87F , T87H , T87W , L88A, L88C , L88H , Q 90N , Q 90W , I91G, I91L, I91M , I91S, L92G , L92N , L92Q , L92S, L92T , L92Y , Q 93P , D94P, V 95F , V 95N , V 95Q , K 96P , A 97 C , A 97P , A 98 P , A 98V , A 100D , A 100E , A 100Q , A 100Y , P 102L, P 102Q , P 102R , L103E , L103K , L104A , L104Q , L104W , L104Y , M 105C , M 105E , M 105F , M 105L , Q 106D , Q 106G , Q 106L , Q 106V , Q 106W , Q 106Y , I107A , I107C , I107E , I107G , I107K, I107L, I107Q , I107S , I107T , R 108G , L109F, L109V , L109Y , P 110A , P 110E , P 110F , P 110G , P 110H , P 110N , P 110S , P 110V , A 111Y , N 112F, N 112P , Y 113D , Y 113E , Y 113P , R 115W , Y 117A , Y 117D , Y 117E , Y 117G , Y 117P , Y 117Q , N 118F , E 119P , A 120P , F121C , F121L, F121M , F121N , F121Q , F121R , F121V , F121W , F121Y , S 122D , S 122F , S 122L , S 122P , S 122W , S 122Y , I124A , I124G , I124H , I124K , I124R , K 127P , L128S , A 129I, A 129W , A 129Y , K 130P, L136A , L136D , L136E , L136G , L136K , L136N , L136P , L136Q , L136S , L136T , L137A , L137C , L137H , L137K , L137Q , L137S , L137Y , P 138F , F139L, F139M , F140C , F140I, F140L, F140M , F140V , M 141T , E 143P , V 144H , Y 145I, L146G , L146P , W 150 G , W 150 I, W 150 V , M 151F , M 151L , M 151R , M 151S , M 151T , M 151W , Q 152N , Q 152V , Q 152Y , D 154C , D 154E , G 155 I, I156C , I156K , I156T , I156V , P 158G , P 158T , A 162T , Q 163A , Q 163C , Q 163E , Q 163 I, Q 163S , Q 163T , Q 163V , I166C , A 167E , A 167F , A 167L , A 167N , A 167R , A 167V , A 167Y , W 169 K , M 170N , M 170S , Q 173D , L174A , L174 T y /o L174W , en donde, en las d e s ig n a c io n e s de m u tac ion po r sus titu c ion , los nu m e ro s se re fie ren a p o s ic io ne s de a m in o a c id o s de la S E C ID NO: 31. Un de riva d o de a c id o g ra so e je m p la r que se p ro d u ce de a cu e rd o con e s to es un a c id o g ra so libre. El a u m e n to y /o la m e jo ra de la p ro du cc ion de un de riva d o de ac ido g ra so pu ede m ed irse in v itro y /o in v ivo . P re fe re n tem e n te , el au m e n to y /o la m e jo ra de la p ro du cc ion de un de riva d o de ac ido g ra so se m id e in vivo .

In one aspect, it is only one uncle is te ra sa mu ta n te (or one of his eq u iva le n te s na tu ra rales), which is the capital of the increase and omega ra la For one or more of the two years of succession, and that is a variant of an uncle, it is known that a co-re r This is done by SEQ ID NO: 31 and x in this in FIG. 58, where the uncle is the one who knows with one or more of his titles, he is given the following: D 2L, D 2P, D2R, L5G, L11I, S 12N, S 12T, A 13N, G 14C, G 14P, G 14S, G 14T, G 14V, Y 15C, Y 15I, Y 15V, R16T, M 17D, M 17E, M 17N, M 17R, M 17S, M 17V, A 19C, A 21G, A 22 L, A 22R, A 22 T, A 25P, L26D, L26G, L26W, L27C, L27F, L27W, L27Y, N28I, N28P, D29P, K30P, W 31D, W 31G, W 31N , W 31P, W 31R, W 31S, W 31T, V 37Y, N 39P, S 41C, I42D, I42G, S43E, G 44K, G 44R, G 44W, D 45G, Q 49E, G 50A, G 50K, G 50M, G 50Q, L51D, L51T, R 53A, R 53G, R 53L, R 53N, R53S, R 53V, L54E, L54F, L54G, L54N, L54S, L54W, L58R, P63G, P 63M, P63N, P 63T, P 63W, W 65E, W 65G, V 66G, V 66S, L67T, V 68S, E 69F, E 69V, L70C, L70F, L70Q, L70S, L70T, L70V, G 71A, N 73G, N 73L, D 74A, D 74C, G 75A, G 75C, G 75F, G 75R, G 75W, L76I, R 77A, R 77C, R 77D, R77F, R 77G, R 77H, R 77K, R 77L, R 77N, R 77Q, R 77S, R 77 W, G 78D, G 78E, F79K, Q 80G, T84H, T 84N, T84Q, T87A, T87F, T87H, T87W, L88A, L88C, L88H, Q 90N, Q 90W, I91G, I91L, I91M, I91S, L92G, L92N, L92Q, L92S, L92T, L92Y, Q 93P, D94P, V 95F, V 95N, V 95Q, K 96P, A 97 C, A 97P, A 98 P, A 98V, A 100D, A 100E, A 100Q, A 100Y, P 102L, P 102Q, P 102R, L103E, L103K, L104A, L104Q, L104W, L104Y, M 105C, M 105E, M 105F, M 105L, Q 106D, Q 106G, Q 106L, Q 106V, Q 106W , Q 106Y, I107A, I107C, I107E, I107G, I107K, I107L, I107Q, I107S, I107T, R 108G, L109F, L109V, L109Y, P 110A, P 110E, P 110F, P 110G, P 110H, P 110N, P 110S, P 110V, A 111Y, N 112F, N 112P, Y 113D, Y 113E, Y 113P, R 115W, Y 117A, Y 117D, Y 117E, Y 117G, Y 117P, Y 117Q, N 118F, E 119P, A 120P, F121C, F121L, F121M, F121N, F121Q, F121R, F121V, F121W, F121Y, S 122D, S 122F, S 122L, S 122P, S 122W, S 122Y, I124A, I124G, I124H, I124K, I124R, I124R 127P, L128S, A 129I, A 129W, A 129Y, K 130P, L136A, L136D, L136E, L136G, L136K, L136N, L136P, L136Q, L136S, L136T, L137A, L137C, L137H, L137K, L137Q, L137S, L137Y, P 138F, F139L, F140, F1, F1, F1, F140, F1, F1 , M 141T, E 143P, V 144H, Y 145I, L146G, L146P, W 150 G, W 150 I, W 150 V, M 151F, M 151L, M 151R, M 151S, M 151T, M 151W, Q 152N, Q 152V, Q 152Y, D 154C, D 154E, G 155 I, I156C, I156K, I156T, I156V, P 158G, P 158T, A 162T, Q 163A, Q 163C, Q 163E, Q 163 I, Q 163S, Q 163T, Q 163V, I166C, A 167E, A 167F, A 167L, A 167N, A 167R, A 167V, A 167Y, W 169 K, M 170N, M 170S, Q 173D, L174A, L174 T and / or L174W, where, in the mutational decisions by their substitution, the numbers will be recognized as part of the provisions of SEQ ID NO: 31 One of riva do de c id og ra so e je mp la r p p du du de a cu e rd o with this is a free id. The increase and / or improvement of the protection of a major asset can only go live and / or live. P re fe re n tem in te, the increase and / or the improvement of the protection of a riva do de r ado g ra so se m id and in vivo.

En un asp ec to , se d e sve la una t io e s te ra s a m u ta n te (o uno de sus e q u iv a le n te s na tu ra les), que es ca p a z de p ro d u c ir un a u m e n to de l re n d im ie n to p ro p o rc io n a l o p o rce n tu a l de d e riv a d o s de a c id o s g ra so s de ca d e n a co rta (p o r e jem p lo , C e, C g, C 10, C 11, C 12, C 13, C 14) (p o r e jem p lo , ac ido s g ra sos de cad en a corta , es te res g ra sos de cad en a corta, a lco h o le s g ra so s de ca d e n a corta , e tc .) fre n te a o tros p ro du c to s (p o r e jem p lo , d e riva d o s de ac ido s g ra sos de cad en a no corta, inc luyendo , po r e jem p lo , ac ido s g ra sos de ca d e n a la rga (p o r e jem p lo , C 15, C 16, C 17, C 18, C 19, C 20), es te res g ra sos de cad en a la rga , a lcoh o le s g ra sos de cadena larga, e tc.), y que es una va ria n te de una t io e s te ra sa p re cu rso ra que co m p re n d e una se cu e n c ia a n a loga a la S E C ID NO: 31 exp u e s ta en la FIG. 58, en dond e la t io e s te ra s a p re cu rso ra se m u ta en una o m as p o s ic io n e s de a m in o a c id o s c o rre s p o n d ie n te s a uno o m as res tos de la S E C ID NO: 31 se le cc io n a d o s de 13, 16-17, 25 -38 , 55 -67 , 78 -98 , 105-119 , 122, 126, 132-145, 153 y/o 161-182. Un de riva d o de ac ido g ra so de ca d e n a co rta e je m p la r es un de riva d o de ac ido g ra so C 12. Un de riva d o de ac ido g ra so de cad en a co rta a lte rn a tivo es un de riva d o de ac ido g raso C 14. En d e te rm in a d a s c ircu ns tan c ias , el au m e n to del re n d im ie n to p ro p o rc io n a l o p o rcen tu a l de l de riva do de ac ido g ra so de cad en a co rta puede c o rre la c io n a rse con una d ism in u c io n de l re n d im ie n to p ro p o rc io n a l de de riva dos de ac ido g ra so de ca d e n a la rga . El au m en to de l re n d im ie n to p ro p o rc io n a l o p o rcen tu a l de d e riva d o s de ac ido s g ra sos de cad en a co rta y /o la co rre sp o n d ie n te d ism in u c io n del re n d im ie n to p ro p o rc io n a l o po rcen tu a l de d e riva d o s de ac ido s g ra sos de cad en a la rga puede m e d irse in vitro o in vivo. P re fe re n te m e n te , el a u m e n to de l re n d im ie n to p ro p o rc io n a l de d e riv a d o s de a c id o s g ra so s de ca d e n a co rta o la d ism in u c io n de l re n d im ie n to p ro p o rc io n a l o po rcen tu a l c o rre sp o n d ie n te de d e riva d o s de ac ido s g ra sos de cad en a la rga se m ide in vivo.

In an asp e ct, it is only one uncle that is te sam sam ta n te (or one of its equals to your nation), which is peace of mind to increase From the re nd im ie n to p ro po rc io nalopo rce n tu al de riv ados de c c osg ra so s de ca dena co rta (pore jem p lo, C e, C g, C 10, C 11, C 12, C 13, C 14) (by jem p lo, acted as a matter of cad in a short term, this is a matter of cad in a short term, which is followed by Cana de Corta, e tc.) fre n te ao tros p ro du c to s (pore jem p lo, de riva dos de ac gone s s se ra ra es de cad in a not cut, including, po re jem p lo, sg ra are from ca dena la rga (pore jem p lo, C 15, C 16, C 17, C 18, C 19, C 20), this is the case of cad in a la rga, a lcoh or you are long chain, e tc.), and that it is a variant of an uncle that is why you know that one is similar to the SEQ ID NO: 31 shown in FIG. 58, where the uncle is the one who knows in one or more positions of am sponsorship or sponsorship in one of the others of the SEQ ID NO: 31 were given swims of 13, 16-17, 25 -38, 55 -67, 78 -98, 105-119, 122, 126, 132-145, 153 and / or 161-182. A riva do de ac gone g ra so de ca dena co rta e je mp la r is a riva do de ac a g gone so C 12. A riva do de ac gone g ra so de cad in a corta A rive a tivo is a private riva do c C 14. In te rm in adasc ircu ns tan c ias, the increase of the re nd im ie n to p ro po rc io nalopo rcen tu al From the riva do de ac gone g ra so de cad in a corta can the c rio be born with a d ism in uction of the re r nd im ie n to p ro po r rio io nal de de riva Two of the people from Cana de la Rga. The au m en to de l re nd im ie n to p ro po rc io nalopo rcen tu de de riva dos de ac gone along with a cad in a corta and / or the cor rre sp ond ie n te d Ism in u n io n of the re nd im ie n to p ro po rc io nalo p r r e r r e rce two of ac c a s g ra es de s in ra rga can be measured in vitro or in vivo. P re fe re n te ment, the increase of the re nd im ie n to p ro p rial of the riv ados de c id osg ra so s de ca dena co rta or the d ism in uc io n of l re nd im ie n to p ro po rc io nalo po rc tu alco rre sp ond ie n te de riva two of ac c a se se ra es de cad in a la rga se m ide in vivo.

En un asp ec to , se d e sve la una t io e s te ra s a m u ta n te (o uno de sus e q u iv a le n te s na tu ra les), que es ca p a z de p ro d u c ir un a u m e n to de l re n d im ie n to p ro p o rc io n a l o p o rce n tu a l de d e riv a d o s de a c id o s g ra so s de ca d e n a co rta (p o r e jem p lo , C 8, C g, C 10, C 11, C 12, C 13, C 14) (p o r e jem p lo , ac ido s g ra sos de cad en a corta , es te res g ra sos de cad en a corta, a lco h o le s g ra sos de cad en a corta, e tc .) fre n te a o tros p ro d u c to s (p o r e jem p lo , d e riva d o s de ac ido s g ra sos de cad en a no corta , in c luye nd o , po r e jem p lo , a c id o s g ra so s de ca d e n a la rga , e s te re s g ra so s de ca d e n a la rga , a lco h o le s g ra sos de ca d e n a larga, etc.), y que es una va ria n te de una t io e s te ra sa p re cu rso ra que c o m p re n d e una se cu e n c ia a n a lo g a a la S E C ID NO: 31 exp ue s ta en la FIG. 58, en dond e la t io e s te ra s a p re cu rso ra se m u ta en una o m as po s ic io ne s de a m in o a c id o s co rre s p o n d ie n te s a uno o m as res tos de la S E C ID NO: 31 s e le c c io n a d o s de 13, 16, 17, 25, 29, 31, 35, 36, 38, 55, 57, 58, 59, 61, 62, 63, 64, 65, 66, 67, 78, 79, 82, 83, 84, 85, 86, 87, 89, 90, 93, 94, 95, 96, 97, 98, 105, 106, 108, 111, 113, 114, 117, 119, 122, 126, 132, 135, 136, 139, 142, 144, 145, 153, 161, 162, 165, 168, 173, 175, 176, 178, 179, 180, 181 y/o 182. Un de riva do de ac ido g raso de cad en a co rta e je m p la r es un de riva do de ac ido g ra so C 12. Un de riva d o de ac ido g ra so de cad en a corta a lte rn a tivo es un de riva d o de ac ido g ra so C 14. En d e te rm in a d a s c ircu ns tan c ias , el a u m en to del re n d im ie n to p ro po rc ion a l o po rcen tu a l de de riva dos de ac idos g rasos de cad en a co rta puede c o rre la c io n a rse con una d ism inu c io n del ren d im ien to p ro po rc ion a l de de riva dos de ac idos g ra sos de cad en a larga. El a u m en to del ren d im ien to p ro po rc ion a l o po rcen tu a l de de riva dos de ac ido s g ra sos de cad en a co rta y /o la c o rre sp o n d ie n te d ism in u c io n de l ren d im ien to p ro p o rc io n a l o po rcen tu a l de d e riva d o s de ac idos g ra sos de cad en a la rga puede m ed irse in vitro o in vivo. P re fe re n te m e n te , el au m e n to de l re n d im ie n to p ro po rc ion a l de d e riva d o s de ac ido s g ra sos de cad en a co rta o la d ism in u c io n de l re n d im ie n to p ro po rc ion a l c o rre sp o n d ie n te de d e riva d o s de ac ido s g ra sos de ca d e n a la rga se m ide in vivo.

In an asp e ct, it is only one uncle that is te sam sam ta n te (or one of its equals to your nation), which is peace of mind to increase From the re nd im ie n to p ro po rc io nalopo rce n tu de riv ados de a c id osg ra so s de ca dena co rta (pore jem p lo, C 8, C g, C 10, C 11, C 12, C 13, C 14) (by jem p lo, ac se sg ra es de cad in a short, this res te rars de cad in a short, lco ho sg ra sos de cad in a short, e tc.) fre n te ao tros p ro duc to s (pore jem p lo, de riva dos de acc sg ra are of cad in a not cut, in c luye nd o, po re jem p So, what are you doing at Cana de la Rga, is what you are doing at Cana de la Rga, what are you doing for a long time, etc.), and what is a va An uncle's son is the one who understands that one is still in SEQ ID NO: 31 set forth in FIG. 58, where the uncle is what you know in one of the most important cases or sponsors of one of the other SEQ ID NO: 31 was given 13, 16, 17, 25, 29, 31, 35, 36, 38, 55, 57, 58, 59, 61, 62, 63, 64, 65, 66, 67, 78, 79, 82, 83, 84, 85, 86, 87, 89, 90, 93, 94, 95, 96, 97, 98, 105, 106, 108, 111, 113, 114, 117, 119, 122, 126, 132, 135, 136, 139, 142, 144, 145, 153, 161, 162, 165, 168, 173, 175, 176, 178, 179, 180, 181 and / or 182. Un de riva do of the private sector of cad in a co rta e je mp la r is a riva do de ac gone g ra so C 12. A of riva do de ac gone g ra so de cad in a short to lte rn a tivo it is a riva do de ac ido g ra so C 14. In te rm in adasc ircu ns tan c ias, the increase in the income of the country is possible in the area of riva Two of the acidic acids of cad in a corta can lead to the birth of a division of the national income tax of two of acidic acids. from ca d in long. The increase in the income tax rate is that you can continue to be responsible for each other in a corta and / or the credit card where you are in. You can use two of acidic acids that are present in the body and can be in vitro or in vivo. P re fe re n te ment, the increase of the re nd im ie n to p ro po rc ion to that of de riva two of ac c a s se ra ra es de cad in a corta or the d ism in uc io n of l re nd im ie n to p ro po rc ion alco rre sp ond ie n te de de riva two de ac ag se s ra ra s de ca dena rga se m ide in vivo.

En un asp ec to , se d e sve la una t io e s te ra s a m u ta n te (o uno de sus e q u iv a le n te s na tu ra les), que es ca p a z de p ro d u c ir un a u m e n to de l re n d im ie n to p ro p o rc io n a l o p o rce n tu a l de d e riv a d o s de ac ido s g ra so s de ca d e n a co rta (p o r e jem p lo , C e, C 9, C 10, C 11, C 12, C 13, C 14) (p o r e jem p lo , ac ido s g ra sos de cad en a corta , es te res g ra sos de cad en a corta, a lco h o le s g ra sos de cad en a corta , e tc .) fre n te a o tros p ro d u c to s (p o r e jem p lo , d e riva d o s de ac ido s g ra sos de cad en a no corta inc luyendo , po r e jem p lo , ac ido s g rasos de cad en a la rga , es te re s g rasos de cad en a larga, a lcoh o le s grasos de ca d e n a larga, etc.), y que es una va ria n te de una tio e s te ra s a p re cu rso ra que c o m p re n d e una se cu e n c ia a n a lo g a a la S E C ID NO: 31 exp ue s ta en la FIG. 58, en do n d e la t io e s te ra sa p re cu rso ra se m uta con una o m as sus titu c ion es s e le cc io n a d a s de: A 13V , R 16A , M 17T, A 25S , D 29M , W 31L , T35Y , S 36W , V 38S , P55A , P55G , L57I, L58M , L58V, K59E , H 61W , Q 62M , P63V, R 64M , W 65L , V 66C , L67C , L67M , G 78F , G 78M , G 78R , G 78T , G 78V , F79K , F79Y, Q 82A , Q 82M , Q 82R , Q 83G , Q 83K , T 84M , T 84V , E85A , E85C , E85G , E85Q , E85S , E85T, E85V, E 85W , E85Y, Q 86H , Q 86Y , T 87R , R 89V , Q 90L , Q 93M , Q 93N , Q 93V , D 94C , D 94L, V 95G , K 96C , A 97N , A 97 V , A 98 G , A98Y , M 105I, Q 106K , Q 106R , R 108W , A 111E , A 111N , A 111S , A 111W , A 111Y , Y 113A , Y 113S , Y 113V , G 114K , G 114Y , Y 117R , E 119M , E 119Q , E 119R , S 122F , S 122I, S 122M , S 122R , P 126K , F132C , F132D , F132K , F132L, F132N , F132V , P 135A , P 135E , P 135K , P 135Q , L136H , F139L, E 142W , V 144Y , Y 145A , Y 145C , Y 145D , Y 145E , Y 145G , Y 145 I, Y 145L , Y 145M , Y 145N , Y 145R , Y 145S , Y 145T , D 153K , D 153Q , D 161K , A 162 I, F165K , D 168W , Q 173I, Q 175M , P 176Q , P 176R , P 176V , V 178F , V 178G , V 178L , V 178R , V 178S , V 178T , N 179H , H 180E , H 180P , H 180R , H 180S , H 180V , H 180W , D 181R , D 181T, S 182C , S 182D , S 182 G y/o S 182R , en donde, en las d e s ig n a c io n e s de m u tac ion po r sus titu c ion , los nu m e ro s se re fie ren a p o s ic io n e s de a m in o a c id o s de la S E C ID NO: 31. Un d e riva d o de ac ido g ra so de cad en a co rta e je m p la r es un de riva do de ac ido g ra so C 12. Un de riva do de ac ido g raso de cadena co rta a lte rn a tivo es un de riva d o de ac ido g ra so C 14. En d e te rm in a d a s c ircu n s ta n c ia s , el a u m en to de l ren d im ien to p ro p o rc io n a l o po rcen tu a l de d e riva d o s de ac ido s g ra sos de cad en a co rta puede c o rre la c io n a rse con una d ism in u c io n de l ren d im ie n to p ro po rc ion a l de d e riva d o s de ac ido s g ra sos de cad en a larga. El au m en to del re n d im ie n to p ro p o rc io n a l o po rcen tu a l de d e riva d o s de ac ido s g ra sos de cad en a co rta y /o la co rre sp o n d ie n te d ism in u c io n de l re n d im ie n to p ro p o rc io n a l de d e riva d o s de a c id o s g ra so s de ca d e n a la rga pued e m ed irse in vitro o in vivo. P re fe re n te m e n te , el a u m e n to de l re n d im ie n to p ro p o rc io n a l de d e riv a d o s de a c id o s g ra so s de ca d e n a co rta o la d ism in u c io n de l re n d im ie n to p ro p o rc io n a l co rre sp o n d ie n te de d e riva d o s de ac ido s g ra sos de ca d e n a la rga se m ide in vivo.

In an asp e ct, it is only one uncle that is te sam sam ta n te (or one of its equals to your nation), which is peace of mind to increase From the re nd im ie n to p ro po rc io nalopo rce n tu al de riv ados de ac ido sg ra so s de ca dena co rta (pore jem p lo, C e, C 9, C 10, C 11, C 12, C 13, C 14) (pore jem p lo, ac sg ra es de cad in a short, this re g ra es de cad in a short, lco ho sg ra sos de cad en a short, e tc.) fre n te ao tros p ro duc to s (pore jem p lo, de riva dos de acido sg ra sos de cad en a not cut inc, po re jem p lo, ac gone sg In the long term, this is a long term, long-term, long-term, etc., and that it is a variety of an uncle is sap re sap re WHEREAS A SEQUENCE INCLUDES SEQ ID NO: 31 EXPOSED IN FIG. 58, where the uncle is the one who is killed with one or more of his substitutions is: A 13V, R 16A, M 17T, A 25S, D 29M , W 31L, T35Y, S 36W, V 38S, P55A, P55G, L57I, L58M, L58V, K59E, H 61W, Q 62M, P63V, R 64M, W 65L, V 66C, L67C, L67M, G 78F, G 78M , G 78R, G 78T, G 78V, F79K, F79Y, Q 82A, Q 82M, Q 82R, Q 83G, Q 83K, T 84M, T 84V, E85A, E85C, E85G, E85Q, E85S, E85T, E85V, E 85W, E85Y, Q 86H, Q 86Y, T 87R, R 89V, Q 90L, Q 93M, Q 93N, Q 93V, D 94C, D 94L, V 95G, K 96C, A 97N, A 97 V, A 98 G , A98Y, M 105I, Q 106K, Q 106R, R 108W, A 111E, A 111N, A 111S, A 111W, A 111Y, Y 113A, Y 113S, Y 113V, G 114K, G 114Y, Y 117R, E 119M , E 119Q, E 119R, S 122F, S 122I, S 122M, S 122R, P 126K, F132C, F132D, F132K, F132L, F132N, F132V, P 135A, P 135E, P 135K, P 135Q, L136H, F139L, E 142W, V 144Y, Y 145A, Y 145C, Y 145D, Y 145E, Y 145G, Y 145 I, Y 145L, Y 145M, Y 145N, Y 145R, Y 145S, Y 145T, D 153K, D 153Q, D 161K, A 162 I, F165K, D 168W, Q 173I, Q 175M, P 176Q, P 176R, P 176V, V 178F, V 178G, V 178L, V 178R, V 178S , V 178T, N 179H, H 180E, H 180P, H 180R, H 180S, H 180V, H 180W, D 181R, D 181T, S 182C, S 182D, S 182 G and / or S 182R, where, in With regard to their mutational positions, their numbers are recognized as part of the statements of SEQ ID NO: 31. One of riva do from ac g o g ra so from cad in a co rta e je mp la r is a riva do de ac gone g ra so C 12. A de riva do de ac gone chain satin corta a lte rn a tivo es A riva do de ac gone g ra so C 14. In de te rm in adasc ircu ns ta nc ia s, the increase in the number of people to do it is only possible Two of the acquaintances of each one of them can be made with a term of reference to the income of the de riva two of the accrual. sg ra sos de cad in long. The au m en to of the re nd im ie n to p ro po rc io nalo po r tu tu de riva dos de ac a se g ra are from cad in a corta and / or the cor rre sp ond ie n te d Ism in uc io n of l re nd im ie n to p ro p rio nal de de riva dos de a c id osg ra so s de ca de la rga can go in vitro or in vivo. P re fe re n te ment, the increase of the re nd im ie n to p ro p rial of the riv ados de c id osg ra so s de ca dena co rta or the d ism in uc io n of l re nd im ie n to p ro po rc io nal co rre sp ond ie n te de riva two of ac cg sg ra are from cana de la rga se m ide in vivo.

En un aspec to , se d e sve la una t io e s te ra s a m u ta n te (o uno de sus e q u iva le n te s na tu ra les), que es ca p a z de p ro d u c ir una d ism inu c io n del ren d im ien to p ro p o rc io n a l o po rcen tu a l de de riva dos de ac ido s g ra sos de cad en a co rta (po r e jem p lo , C 8, C 9, C 10, C 11, C 12, C 13, C 14) (p o r e jem p lo , ac ido s g ra sos de cadena corta , e s te re s g ra sos de cadena corta , a lcoh o le s g ra sos de cad en a corta, e tc .) fre n te a o tro s p ro du c to s (p o r e jem p lo , d e riva d o s de ac ido s g ra sos de cad en a no co rta in c luye nd o , po r e jem p lo , ac ido s g rasos de cad en a la rga (p o r e jem p lo , C 15, C 16, C 17, C 18, C 19, C 20), e s te re s g ra sos de cadena larga, a lcoh o le s g rasos de cadena larga, e tc.), y que es una va ria n te de una t io e s te ra sa p re cu rso ra que co m p re n d e una se cu e n c ia a n a loga a la S E C ID NO: 31 exp u e s ta en la FIG. 58, en dond e la t io e s te ra s a p re cu rso ra se m u ta en una o m as p o s ic io n e s de a m in o a c id o s c o rre s p o n d ie n te s a uno o m as res tos de la S E C ID NO: 31 s e le cc io n a d o s de 1-31, 36 -81 , 84 -159 , 162 -177 y /o 181. Un de riva d o de ac ido g ra so de cad en a corta e je m p la r es un de riva d o de ac ido g raso C 12. Un de riva do de ac ido g ra so de cad en a corta a lte rna tivo es un de riva do de a c id o g raso C 14. En d e te rm in a d a s c ircu n s ta n c ia s , la d ism in u c io n de l re n d im ie n to p ro po rc ion a l o po rcen tu a l de d e riva d o s de a c id o s g ra so s de ca d e n a co rta pu ed e co rre la c io n a rs e con un a u m e n to de l re n d im ie n to p ro p o rc io n a l de d e riva d o s de a c id o s g ra so s de ca d e n a larga. La d ism in u c io n de l re n d im ie n to p ro p o rc io n a l o p o rce n tu a l de d e riva d o s de ac ido s g ra sos de cad en a co rta y /o el c o rre sp o n d ie n te a u m e n to de l re n d im ie n to p ro po rc ion a l de de riva dos de ac ido s g ra sos de cad en a la rga puede m ed irse in vitro o in vivo. P re fe re n te m e n te , la d ism in u c io n de l ren d im ie n to p ro p o rc io n a l de d e riva d o s de ac ido s g ra sos de cad en a co rta o el c o rre sp o n d ie n te a u m e n to de l ren d im ien to p ro p o rc io n a l de de riva dos de ac ido s g ra sos de ca d e n a co rta se m ide in vivo.

In one respect, it is only one of these is te sam sam ta n te (or one of its equals le na na ra rales), which is peace of mind to go one day. I n of the ren d im ien to p ro po rc io n r rio io n n r r e r r e nce de riva two of a c a c a c rta (po re jem p lo, C 8, C 9, C 10, C 11, C 12, C 13, C 14) (by jem p lo, ac se sg ra are short chain, s re sg ra are short chain, lcoh or le s ra ra de cad in short , e tc.) fre n te ao tro sp ro du c to s (pore jem p lo, de riva dos de ac a sg ra sos de cad en a no co rta in c luye nd o, po re jem p lo, Here are satos de cad en a la rga (pore jem p lo, C 15, C 16, C 17, C 18, C 19, C 20), this is long chain, to lcoh or le long chain satin, e tc.), and that it is a variant of an uncle that is why he knows what a sequence is similar to SEQ ID NO : 31 shown in FIG. 58, where the uncle is the one who knows in one or more positions of am sponsorship or sponsorship in one of the others of the SEQ ID NO: 31 you will be sworn from 1-31, 36 -81, 84 -159, 162 -177 and / or 181. A riva do de ac gone g ra so de cad in a short e je mp la r is one of riva do de ac a g sato C 12. A of riva do de ac gone g ra so de cad in a short a l r rive tive is a riva do de c a o id sato c 14. In de te rm in Adasc ircu ns ta nc ia s, the d ism in uc io n of l re nd im ie n to p ro po rc ion alo r rr tu tu de riva dos de a c id osg ra so s de ca dena co rta It is possible to run the nation with an increase in the national income of two of the long-term channels. The direction of the re nd im ie n to p ro po rc io nalopo rce n tu de de riva dos de ac gone s es ra ra de cad in a corta and / or the cor spre sp ond ie If you want to increase your income, you can leave it in vitro or in vivo. P re fe re n te ment, the r ism in uc io n of l ren d im ie n to p ro po rc io nal de de riva two of ac c sg ra are of cad in a corta or the cor rre In addition, I am increasing your income from the Federal Republic of the United States, as well as from Canada .

En un aspec to , se d e sve la una t io e s te ra s a m u ta n te (o uno de sus e q u iv a le n te s n a tu ra les ), q u e es ca p a z de p ro d u c ir una d ism inu c io n del ren d im ien to p ro p o rc io n a l o po rcen tu a l de de riva dos de ac ido s g ra sos de cad en a co rta (po r e jem p lo , C 8, C g, C 10, C 11, C 12, C 13, C 14) (p o r e jem p lo , ac ido s g ra sos de cadena corta , e s te re s g ra sos de cadena corta , a lcoh o le s g ra sos de cad en a corta, e tc .) fre n te a o tros p ro d u c to s (p o r e jem p lo , d e riva d o s de ac ido s g ra sos de ca d e n a no co rta in c luye nd o , po r e jem p lo , a c id o s g ra so s de ca d e n a larga, es te re s g ra so s de ca d e n a larga, a lco h o le s g ra sos de cad en a larga, etc.), y que es una va ria n te de una t io e s te ra sa p re cu rso ra que com p ren de una secu en c ia an a lo g a a la S E C ID NO: 31 exp ue s ta en la FIG. 58, en donde la tio e s te ra sa p re cu rso ra se m u ta en una o m as p o s ic io n e s de a m in o a c id o s co rre sp o n d ie n te s a uno o m as res tos de la S E C ID NO: 31 s e le cc io n a d o s de 1, 2, 3, 4, 5, 6 , 7, 8, 9, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21, 22, 23, 24 , 26, 27, 30, 31, 36, 37, 38, 42, 44 , 45, 46, 47, 48, 50, 51, 52, 53, 54, 55, 57, 61, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 81, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 99, 100, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 117, 118, 119, 120, 121, 122, 124, 125, 127, 128, 129, 130, 131, 132, 133, 134, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 162, 163, 165, 166, 167, 168, 170, 171, 173, 174, 175, 176, 177 y /o 181. Un de riva d o de ac ido g ra so de cad en a co rta e je m p la r es un de riva d o de ac ido g ra so C 12. Un de riva d o de ac ido g ra so de cad en a co rta a lte rna tivo es un de riva do de ac ido g ra so C 14. En de te rm in ada s c ircu n s ta n c ia s , la d ism in u c io n de l re n d im ie n to p ro p o rc io n a l o p o rce n tu a l de d e riva d o s de ac ido s g ra sos de cadena co rta pued e co rre la c io n a rs e con un au m e n to de l ren d im ie n to p ro p o rc io n a l de d e riva d o s de a c id o s g ra so s de cad en a la rga . La d ism inu c io n de l re n d im ie n to p ro po rc ion a l o po rcen tu a l de de riva dos de a c id o s g ra so s de cad en a co rta y/o el co rre sp o n d ie n te a u m e n to de l re n d im ie n to p ro po rc ion a l de d e riva d o s de ac ido s g ra sos de cad en a la rga puede m ed irse in vitro o in vivo. P re fe re n te m e n te , la d ism inu c io n de l re n d im ie n to p ro po rc ion a l de d e riva d o s de ac idos g ra sos de cad en a co rta o el c o rre sp o n d ie n te au m e n to de l ren d im ie n to p ro po rc ion a l de d e riva d o s de ac ido s grasos de ca d e n a co rta se m ide in vivo.

In one respect, it is only one of these is tera samu ta n te (or one of his teams), which is the peace of mind to go to a city I n of the ren d im ien to p ro po rc io n r rio io n r r e rce two of ac c a s se ra ra de cad in a corta (po re jem p lo, C 8, C g, C 10, C 11, C 12, C 13, C 14) (by jem p lo, ac se sg ra are short chain, s re sg ra are short chain, lcoh or le s ra ra de cad in short , e tc.) fre n te ao tros p ro duc to s (pore jem p lo, de riva dos de ac gone s s ra ra de ca dena no co rta in c luye nd o, po re jem p lo, a c id osg ra so s of long channel, this re sg ra so s of long channel, lco ho sg ra are of long term, etc.), and that is a variant of a t This is the reason why they buy from a section in addition to SEQ ID NO: 31 set forth in FIG. 58, where the uncle is the one who is in one or more positions in the United States or one of the SEC's other authorities. ID NO: 31 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21, 22, 23, 24, 26, 27, 30, 31, 36, 37, 38, 42, 44, 45, 46, 47, 48, 50, 51, 52, 53, 54, 55, 57, 61, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 81, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 99, 100, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 117, 118, 119, 120, 121, 122, 124, 125, 127, 128, 129, 130, 131, 132, 133, 134, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 162, 163, 165, 166, 167, 168, 170, 171, 173, 174, 175, 176, 177 and / or 181. Un de riva do of ac g o ra so de cad in a co rta e je mp la r is a riva do de ac gone g ra so C 12. A de riva do de ac gone g ra so de cad in a corta a lte rna tivo is un of riva do de ac gone g ra so C 14. In de te rm in ada sc ircu ns ta nc ia s, the d ism in uc io n of l re nd im ie n to p ro po rc io nalopo rce n Your de riva dos de ac gone sg ra are from the co rrta cord e la cor io na rs e with an increase of the ren d im ie n to p ro po rc io nal de de riva dos de Here are the cadres of the country. The direction of the re nd im ie n to p ro po rc ion alo r r r e r th e r th e r th of two in a co rt and c o sp sp c ond ie n t l Aumen to re nd im e n t p ro rc po ion to the riva of two ac been sg sos ra ch in the RGA can leave m ed in vitro or in vivo. P re fe re n te ment, the d ism inu tio n of the re nd im ie n to p ro po rc ion to the de riva two of acidic acids of cad in a corta or the cor rre In addition to the development of the riva two of the fatty acids in Canada, the same is in vivo.

En un asp ec to , se d e sve la una t io e s te ra s a m u ta n te (o uno de sus e q u iv a le n te s na tu ra les ), que es ca p a z de p ro d u c ir una d ism inu c io n del ren d im ien to p ro p o rc io n a l o po rcen tu a l de de riva dos de ac ido s g ra sos de cad en a co rta (po r e jem p lo , C e, C 9, C 10, C 11, C 12, C 13, C 14) (p o r e jem p lo , ac ido s g ra sos de cadena corta , e s te re s g ra sos de cadena corta , a lcoh o le s g ra sos de cad en a corta, e tc .) fre n te a o tros p ro d u c to s (p o r e jem p lo , d e riva d o s de ac ido s g ra sos de ca d e n a no co rta in c luye nd o , po r e jem p lo , ac ido s g ra sos de ca d e n a larga, es te re s g ra so s de ca d e n a larga, a lco h o le s g ra sos de cad en a larga, e tc.), y que es una va ria n te de una t io e s te ra sa p re cu rso ra que com p ren de una secu en c ia an a lo g a a la S E C ID NO: 31 exp ue s ta en la FIG. 58, en donde la tio e s te ra s a p re cu rso ra se m u ta con una o m as su s titu c io n e s se le cc io n a d a s de: A 1C , A 1F , A 1L, A 1Y , D2L, D 2M , D2P, D 2W , T3R , L4A, L4M, L4N , L4S, L4V, L4Y, L5E, L5F, L5G, L5K, L5N, L5S, L5W , I6T, L7A, L7E, L7K, L7M , L7W , G 8K, D9N, D9T, L11A , L11C , L11I, L11M , L11Q , L11V, S 12I, S12L, S 12M , S 12N , S 12T , S 12V , S 12Y , A 13 C , G 14 C , G 14E , G 14 I, G 14M , G 14N , G 14P , G 14S , G 14T , G 14V , Y 15C , Y 15E , Y 15G , Y 15I, Y 15N , Y 15V , R 16T, M 17D , M 17E , M 17G , M 17L, M 17N , M 17P , M 17R , M 17S , M 17V , S18M , S 18N , S 18T , A 19E , A 19L , A 19 V , A 21 P , A 22 D , A 22E , A 22 F , A 22H , A 22 I, A 22 K , A 22L, A 22P , A 22 R , A 22 S , A 22T , A 22 Y , W 23A , W 23H , W 23N , W 23P , P24A , P24C , P24D , P24E, P 24F, P 24G , P24I, P 24M , P24N , P24S , P 24T, P24V, P 24W , L26P , L27A, L27C , L27F, L27H , L27R , L27S , L27T , L27W , L27Y, K 30P , W 31 D , W 31P , W 31R , S 36F, S 36L, V 37G , V 37H , V 37N , V 37Q , V 37 W , V 37Y , V 38P , N39E, N 39G , N39K, N 39M , N39P, N39Q , N 39Y , I42D , I42G , I42P, G 44A , G 44E , G 44K , G 44M , G 44N , G 44R , G 44S , G 44W , G 44Y , D 45G , D 45M , T 46D , S 47E , S 47P , S 47Q , S47R , S 47Y , Q 48Y , G 50C , G 50E , G 50F , G 50I, G 50K , G 50L, G 50M , G 50N , G 50P , G 50Q , G 50R , G 50S , G 50T , G 50W , G 50Y , L51D , L51P, L51T, A 52 P , R53A, R53C , R53D , R 53E , R53F, R53G , R53I, R 53K , R53L, R 53N , R53S, R53T, R 53V , R 53W , R53Y, L54C , L54E , L54G , L54N , L54Y, P 55Y , L57P, H61A, H 61D , H61E, P63D , P63E , P63G , P63K, P 63M , P63N , P63Q , P63R , R 64L, W 65 G , W 65P , W 65R , V 66N , V 66Q , V 66S , V 66W , V66Y , L67E , L67G , L67Q , L67R , L67S, L67W , V 68E , V 68G , V 68N , V 68P , V 68Q , E69A, E 69C , E69D, E 69F, E69G , E69H , E 69K , E69L, E69M , E 69N , E69P , E69Q , E69S , E 69V , E 69W , E69Y , L70A , L70C , L70E , L70F, L70G , L70H , L70K, L70Q , L70S , L70T , L70W , G 71C , G 71S , G 72A , G 72M , G 72P , N 73A , N 73G , N 73H , N73I, N 73L , N 73P , N 73R , N 73S , N 73T , N 73W , D74A, D74C , D74F, D 74G , D74Q , D74S, D 74W , D74Y, G 75A , G 75C , G 75D , G 75E , G 75F , G 75I, G 75K , G 75L, G 75M , G 75N , G 75P , G 75R , G 75T , G 75V , G 75W , G 75Y , L76A, L76C , L76D , L76E, L76F, L76G , L76I, L76K , L76M , L76N , L76P, L76Q , L76R , L76T , L76V , L76W , R 77A , R 77C , R77D , R 77E , R 77F, R 77G , R 77H , R77N , R 77S , R 77V , R 77W , G 78A , G 78C , G 78D , G 78E , G 78N , G 78P , G 78Q , G 78Y , F79P , F79Q , F79S , F79V , P81E, P 81W , T 84D , T84E , T84G , T84H , T84K , T84L , T84N , T84Q , T 84R , T84W , T84Y , E85F, E85P , Q 86A , T87F , L88A, L88E , L88G , L88H , L88Q , L88S , L88W , L88Y , R 89P , Q 90P , Q 90W , I91E, I91L, I91M , I91N , I91Q , I91S, I91Y, L92C , L92E , L92G , L92H , L92N , L92Q , L92R , L92S , L92Y , Q 93P , D 94P , D 94V , V 95A , V 95C , V 95D , V 95E , V 95F , V95I, V 95P , V 95Q , V 95W , V 95Y , K96P , A 97C , A 97 P , N99D , A 100Q , A 100Y , P 102E , P 102G , P 102H , P102L, P 102R , P 102V , P 102W , L103C , L103E , L103I, L103K , L103N , L103R , L103S , L103T , L103V , L104A , L104C , L104E , L104G , L104I, L104N , L104P , L104Q , L104S , L104W , L104Y , M 105A , M 105C , M 105E , M 105F , M 105G , M 105K , M 105L, M 105P , M 105T , M 105W , Q 106D , Q 106G , Q 106H , Q 106L , Q 106W , I107A , I107E , I107F, I107G , I107K , I107L, I107Q , I107S , I107T , I107Y , R 108A , R 108C , R 108D , R 108E , R 108F, R 108G , R 108H , R 108I, R 108L, R 108M , R 108S , R 108V , R 108Y , L109C , L109F, L109G , L109K , L109Q , L109R , L109T, L109V , L109Y , P 110A , P 110C , P 110D , P 110E , P 110F , P 110G , P 110H , P 110K , P110L, P 110M , P 110N , P 110R , P 110S , P 110V , P 110W , A 111C , A 111L , A 111P , A 111Q , A 111R , A 111V , N 112I, N 112L, N 112P , N 112Y , Y 113D , Y 113E , Y 113Q , G 114A , R 115W , Y 117D , Y 117G , Y 117P , N 118F, E 119C , E119L, A 120P , F121A , F121C , F121D , F121E , F121G , F121K , F121L, F 121N , F121P , F121Q , F121R , F121S , F121V , F121W , F121Y , S 122D , S 122E , S 122L , S 122P , I124D , I124E, I124G , I124H , I124K , I124R , I124W , I124Y , Y 125C , Y 125G , Y 125H , Y 125 I, Y 125L , Y 125P , Y 125Q , Y 125R , Y 125S , Y 125T , Y 125V , K 127A , L128E , L128F, L128G , L128K , L128Q , L128R , L128S , L128W , A 129D , A 129F , A 129L , A 129 W , A 129Y , K 130P , K 130V , E 131A , E 131C , E 131D , E 131P , E 131V , F132P , D 133C , V 134C , V 134D , V 134N , V134P, V134W, L136A, L136D, L136E, L136G, L136N, L136P, L136T, L137D, L137E, L137G, L137H, L137K, L137P, L137Q, L137R, L137S, P138G, P138N, P138V, F139A, F139C, F139D, F139E, F139G, F139H, F139M, F139N, F139S, F139T, F139V, F139W, F140A, F140C, F140G, F140I, F140L, F140M, F140N, F140P, F140S, F140T, F140V, F140W, M141C, M141D, M141E, M141F, M141G, M141K, M141L, M141P, M141Q, M141R, M141T, M141W, M141Y, E142A, E142C, E142G, E142I, E142L, E142M, E142P, E142Q, E142R, E142T, E142V, E143A, E143D, E143F, E143G, E143I, E143M, E143P, E143W, V144A, V144D, V144E, V144G, V144H, V144N, V144P, V144Q, V144R, V144S, Y145Q, Y145W, L146C, L146P, W150P, W150R, M151A, M151C, M151D, M151E, M151F, M151G, M151I, M151L, M151Q, M151R, M151S, M151T, M151V, M151W, Q152P, D153A, D153E, D153F, D154A, D154C, D154E, D154F, D154G, D154H, D154I, D154K, D154L, D154M, D154N, D154P, D154R, D154S, D154T, D154V, D154W, G155A, G155P, G155V, I156A, I156C, I156E, I156F, I156G, I156K, I156M, I156Q, I156R, I156S, I156T, I156Y, H157C, H157E, P158F, P158H, P158I, P158L, P158Q, P158V, P158W, N159P, N159W, A162K, A162L, A162N, A162R, A162Y, Q163A, Q163D, Q163E, Q163F, Q163I, Q163V, Q163W, Q163Y, F165L, I166A, I166F, I166M, I166S, I166Y, A167C, A167D, A167E, A167F, A167L, A167N, A167R, A167V, A167W, A167Y, D168M, D168R, M170E, M170F, M170G, M170N, M170S, M170T, A171S, Q173D, Q173P, L174A, L174G, L174S, L174T, L174W, L174Y, Q175F, P176L, P176Y, L177F, L177M, L177S, D181C, D181E y/o D181G, en las que, en las designaciones de mutacion por sustitucion, los numeros se refieren a posiciones de aminoacidos de la s Ec ID NO: 31. Un derivado de acido graso de cadena corta ejemplar es un derivado de acido graso C12. Un derivado de acido graso de cadena corta alternativo es un derivado de acido graso C14. En determinadas circunstancias, la disminucion del rendimiento proporcional o porcentual de derivados de acidos grasos de cadena corta puede correlacionarse con un aumento del rendimiento proporcional de derivados de acidos grasos de cadena larga. La disminucion del rendimiento proporcional o porcentual de derivados de acidos grasos de cadena corta y/o el correspondiente aumento del rendimiento proporcional de derivados de acidos grasos de cadena larga puede medirse in vitro o in vivo. Preferentemente, la disminucion del rendimiento proporcional de derivados de acidos grasos de cadena corta o el correspondiente aumento del rendimiento proporcional de derivados de acidos grasos de cadena corta se mide in vivo.

In an asp e ct, it is only one uncle that is te sam sam ta n te (or one of its equals in your country), which is peace of mind to go to a city Inu c io n of the ren d im ien to p ro po rc io nalo po r te tu de de riva two of ac c a se g ra es de cad in a co rta (po re jem p lo, C e, C 9, C 10, C 11, C 12, C 13, C 14) (by jem p lo, ac se sg ra are short chain, s re sg ra are short chain, lcoh or le s ra ra de cad in a short, e tc.) fre n te ao tros p ro duc to s (pore jem p lo, de riva dos de ac ido sg ra are de ca dena no corta in c luye nd o, po re jem p lo, Here you are from long-term, this is from long-term, and from now on from long-term, etc.), and that is a variant of a This is the reason why they buy from a section in addition to SEQ ID NO: 31 set out in FIG. 58, where the uncle is te sap re when he is changed with one or more of his substitutions: A 1C, A 1F, A 1L, A 1Y, D2L, D 2M , D2P, D 2W, T3R, L4A, L4M, L4N, L4S, L4V, L4Y, L5E, L5F, L5G, L5K, L5N, L5S, L5W, I6T, L7A, L7E, L7K, L7M, L7W, G 8K, D9N , D9T, L11A, L11C, L11I, L11M, L11Q, L11V, S 12I, S12L, S 12M, S 12N, S 12T, S 12V, S 12Y, A 13 C, G 14 C, G 14E, G 14 I, G 14M, G 14N, G 14P, G 14S, G 14T, G 14V, Y 15C, Y 15E, Y 15G, Y 15I, Y 15N, Y 15V, R 16T, M 17D, M 17E, M 17G, M 17L , M 17N, M 17P, M 17R, M 17S, M 17V, S18M, S 18N, S 18T, A 19E, A 19L, A 19 V, A 21 P, A 22 D, A 22E, A 22 F, A 22H, A 22 I, A 22 K, A 22L, A 22P, A 22 R, A 22 S, A 22T, A 22 Y, W 23A, W 23H, W 23N, W 23P, P24A, P24C, P24D, P24E , P 24F, P 24G, P24I, P 24M, P24N, P24S, P 24T, P24V, P 24W, L26P, L27A, L27C, L27F, L27H, L27R, L27S, L27T, L27W, L27Y, K 30P, W 31 D , W 31P, W 31R, S 36F, S 36L, V 37G, V 37H, V 37N , V 37Q, V 37 W, V 37Y, V 38P, N39E, N 39G, N39K, N 39M, N39P, N39Q, N 39Y, I42D, I42G, I42P, G 44A, G 44E, G 44K, G 44M, G 44N, G 44R, G 44S, G 44W, G 44Y, D 45G, D 45M, T 46D, S 47E, S 47P, S 47Q, S47R, S 47Y, Q 48Y, G 50C, G 50E, G 50F, G 50I, G 50K, G 50L, G 50M, G 50N, G 50P, G 50Q, G 50R, G 50S, G 50T, G 50W, G 50Y, L51D, L51P, L51T, A 52 P, R53A, R53C, R53D , R 53E, R53F, R53G, R53I, R 53K, R53L, R 53N, R53S, R53T, R 53V, R 53W, R53Y, L54C, L54E, L54G, L54N, L54Y, P 55Y, L57P, H61A, H 61D, H61E, P63D, P63E, P63G, P63K, P 63M, P63N, P63Q, P63R, R 64L, W 65 G, W 65P, W 65R, V 66N, V 66Q, V 66S, V 66W, V66Y, L67E, L67G, L67Q, L67R, L67S, L67W, V 68E, V 68G, V 68N, V 68P, V 68Q, E69A, E 69C, E69D, E 69F, E69G, E69H, E 69K, E69L, E69M, E 69N, E69P, E69Q , E69S, E 69V, E 69W, E69Y, L70A, L70C, L70E, L70F, L70G, L70H, L70K, L70Q, L70S, L70T, L70W, G 71C, G 71S, G 72A, G 72M, G 72P, N 73A, N 73G, N 73H, N73I, N 73L, N 73P, N 73R, N 73S, N 73T, N 73W, D74A, D74C, D74F, D 74G, D74Q, D74S, D 74W, D74Y, G 75A, G 75C, G 75D, G 75E, G 75F, G 75I, G 75K, G 75L, G 75M, G 75N, G 75P, G 75R, G 75T, G 75V, G 75W, G 75Y, L76A, L76C, L76D , L76E, L76F, L76G, L76I, L76K, L76M, L76N, L76P, L76Q, L76R, L76T, L76V, L76W, R 77A, R 77C, R77D, R 77E, R 77F, R 77G, R 77H, R77N, R 77S, R 77V, R 77W, G 78A, G 78C, G 78D, G 78E, G 78N, G 78P, G 78Q, G 78Y, F79P, F79Q, F79S, F79V, P81E, P 81W, T 84D, T84E, T84G, T84H, T84K, T84L, T84N, T84Q, T 84R, T84W, T84Y, E85F, E85P, Q 86A, T87F, L88A, L88E, L88G, L88H, L88Q, L88S, L88W, L88Y, R 89P, Q 90P, Q 90W, I91E, I91L, I91M, I91N, I91Q, I91S, I91Y, L92C, L92E, L92G, L92H, L92N, L92Q, L92R, L92S, L92Y, Q 93P, D 94P, D 94V, V 95A, V 95C, V 95D, V 95E, V 95F, V95I, V 95P, V 95Q, V 95W, V 95Y, K96P, A 97C, A 97 P, N99D, A 100Q, A 100Y , P 102E, P 102G, P 102H, P102L, P 102R, P 102V, P 102W, L103C, L103E, L103I, L103K, L103N, L103R, L103S, L103T, L103V, L104A, L104C, L104E, L104G, L104G, L104G, L104G , L104P, L104Q, L104S, L104W, L104Y, M 105A, M 105C, M 105E, M 105F, M 105G, M 105K, M 105L, M 105P, M 105T, M 105W, Q 106D, Q 106G, Q 106H, Q 106L, Q 106W, I107A, I107E, I107F, I107G, I107K, I107L, I107Q, I107S, I107T, I107Y, R 108A, R 108C, R 108D, R 108E, R 108F, R 108G, R 108H, R 108I, R 108L, R 108M, R 108S, R 108V, R 108Y, L109C, L109F, L109G, L109K, L109Q, L109R, L109T, L109V, L109Y, P 110A, P 110C, P 110D, P 110E, P 110F, P 110G , P 110H, P 110K, P110L, P 110M, P 110N, P 110R, P 110S, P 110V, P 110W, A 111C, A 111L, A 111P, A 111Q, A 111R, A 111V, N 112I, N 112L , N 112P, N 112Y, Y 113D, Y 113E, Y 113Q, G 114A, R 115W, Y 117D, Y 117G, Y 117P, N 118F, E 119C, E119L, A 120P, F121A, F121C, F121D, F121E, F121G, F121K, F121L, F 121N, F121P, F121Q, F121R, F121S, F121V, F121W, F121Y, S 122D, S 122E, S 122L, S 122P, I124D, I124E, I124G, I124H, I124K, I124R, I124W, I124Y, Y 125C, 125C, Y 125C Y 125H, Y 125 I, Y 125L, Y 125P, Y 125Q, Y 125R, Y 125S, Y 125T, Y 125V, K 127A, L128E, L128F, L128G, L128K, L128Q, L128R, L128S, L128W, A 129D, A 129F, A 129L, A 129 W, A 129Y, K 130P, K 130V, E 131A, E 131C, E 131D, E 131P, E 131V, F132P, D 133C, V 134C, V 134D, V 134N, V134P, V134W, L136A, L136D, L136E, L136G, L136N, L136P, L136T, L137D, L137E, L137G, L137H, L137K, L137P, L137Q, L137R, L137S, P138G, P138N, P138V, F139A, F139C, F139D, F139E, F139G, F139H, F139M, F139N, F139S, F139T, F139V, F139W, F140A, F140C, F140G, F140I, F140L, F140M, F140N, F140P, F140S, F140T, F140V, F140W, M141C, M141D1, M141D1, F149D M141K, M141L, M141P, M141Q, M141R, M141T, M141W, M141Y, E142A, E142C, E142G, E142I, E142L, E142M, E142P, E142Q, E142R, E142T, E142V, E143D3, E143D3, E143D, E143D3, E143D, E143D, E143D3 E143P, E143W, V144A, V144D, V144E, V144G, V144H, V144N, V144P, V144Q, V144R, V144S, Y145Q, Y145W, L146C, L146P, W150P, W150R, M151A, M151C, M151D, M151E, M151F, M151G, M151i, M151L, M151Q, M151R, M151S, M151T, M151V, M151W, Q152P, D153A, D153E, D153F, D154A, D154C, D154E, D154F, D154G, D154H, D154I, D154K, D154L, D154M, D154N, D154P, D154R, D154S, D154T, D154V, D154W, G155A, G155P, G155V, I156A, I156C, I156E, I156F, I156G, I156K, I156M, I156Q, I156R, I156S, I156T, I156Y, H157C, H157E, P158F, P158H, P158I, P158L, P158Q, P158V, P158W, N159P, N159W, A162K, A162L, A162N, A162R, A162Y, Q163A, Q163D, Q163E, Q163F, Q163I, Q163V, Q163W, Q163Y, F165L, I166A, I166F, I166M, I166S, I166Y, A167C, A167D, A167E, A167F, A167L, A167N, A167R, A167V, A167W, A167Y, D168M, D168R, M170E, M170F, M170G, M170N, M170S, M170T, A171S, Q173D, Q173P, L174A, L174G, L174S, L174T, L174W, L174Y, Q175F, P176L, P176Y, L177F, L177M, L177S, D181C, D181E and / or D181G, in which, in the designations of mutation by substitution, at amino acid positions of s Ec ID NO: 31. An exemplary short chain fatty acid derivative is a C12 fatty acid derivative. An alternative short chain fatty acid derivative is a C14 fatty acid derivative. In certain circumstances, the decrease in the proportional or percentage yield of short chain fatty acid derivatives may be correlated with an increase in the proportional yield of long chain fatty acid derivatives. The decrease in the proportional or percentage yield of short chain fatty acid derivatives and / or the corresponding increase in the proportional yield of long chain fatty acid derivatives can be measured in vitro or in vivo. Preferably, the decrease in the proportional yield of short chain fatty acid derivatives or the corresponding increase in the proportional yield of short chain fatty acid derivatives is measured in vivo.

En un aspecto, se proporciona un polinucleotido (o un gen) que codifica una tioesterasa mutante (o uno de sus equivalentes naturales) de la divulgacion. En otro aspecto, se desvela un vector que comprende el polinucleotido (o el gen) segun la presente divulgacion.

In one aspect, a polynucleotide (or a gene) encoding a mutant thioesterase (or one of its natural equivalents) of the disclosure is provided. In another aspect, a vector comprising the polynucleotide (or the gene) according to the present disclosure is disclosed.

En un aspecto, la tioesterasa precursora esta codificada por un gen que puede hibridarse selectivamente con la secuencia de polinucleotidos de ’tesA, o con un ortologo, paralogo u homologo de la misma. En la FIG. 56 se ofrece un listado de Numeros de Registro de GenBank de homologos de proteinas de 'TesA que tienen una identidad de secuencia de aminoacidos de al menos 40% con 'TesA. La tioesterasa precursora puede codificarse mediante un polinucleotido que puede hibridarse selectivamente en condiciones de rigurosidad intermedia, alta o maxima.

In one aspect, the precursor thioesterase is encoded by a gene that can selectively hybridize with the polynucleotide sequence of 'tesA, or with an ortholog, paralogue or homologue thereof. In FIG. 56 A list of GenBank Registration Numbers of 'TesA protein counterparts having an amino acid sequence identity of at least 40% with' TesA is offered. The precursor thioesterase can be encoded by a polynucleotide that can selectively hybridize under conditions of intermediate, high or maximum stringency.

En un aspecto, se desvela un polinucleotido que codifica una tioesterasa precursora en donde la tioesterasa precursora comprende la secuencia de aminoacidos de 'TesA, un ortologo de la misma, un paralogo de la misma o un homologo de la misma. Por ejemplo, la tioesterasa precursora comprende la secuencia de aminoacidos de una 'TesA obtenida de una bacteria E. coli, tal como una bacteria E.coli K12. En un aspecto particular, se desvela un polinucleotido que codifica la tioesterasa precursora, en donde la tioesterasa precursora comprende la secuencia de aminoacidos, una variante, o un fragmento de la SEC ID NO: 31 de la FIG. 58. En un aspecto particular, el gen que codifica la tioesterasa precursora comprende la secuencia de polinucleotidos de la SEC ID NO: 32 expuesta en la FIG. 59, o un fragmento de la misma.

In one aspect, a polynucleotide that encodes a precursor thioesterase is disclosed in which the precursor thioesterase comprises the amino acid sequence of 'TesA, an ortholog thereof, a paralog of the same or a homologue thereof. For example, the precursor thioesterase comprises the amino acid sequence of a 'TesA obtained from an E. coli bacterium , such as an E.coli K12 bacterium . In a particular aspect, a polynucleotide encoding the precursor thioesterase is disclosed, wherein the precursor thioesterase comprises the amino acid sequence, a variant, or a fragment of SEQ ID NO: 31 of FIG. 58. In a particular aspect, the gene encoding the precursor thioesterase comprises the polynucleotide sequence of SEQ ID NO: 32 set forth in FIG. 59, or a fragment thereof.

En un aspecto, se desvela un polinucleotido que codifica una tioesterasa precursora en donde la tioesterasa precursora comprende una proteina que tiene al menos aproximadamente 20 %, por ejemplo, al menos aproximadamente 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 96 %, 97 %, 98 %, 99% o 100% de identidad con la secuencia SEQ ID NO: 31 de la FIG.

In one aspect, a polynucleotide encoding a precursor thioesterase is disclosed wherein the precursor thioesterase comprises a protein having at least about 20%, for example, at least about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% , 99% or 100% identity with the sequence SEQ ID NO: 31 of FIG.

58. En un aspecto, se desvela un polinucleotido que codifica una tioesterasa precursora en donde la tioesterasa precursora comprende una proteina que tiene al menos aproximadamente 20 %, por ejemplo, al menos aproximadamente 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 96 %, 97 %, 98 %, 99 % o 100 % de identidad con la secuencia de una 'TesA de la bacteria E.coli K12. En un aspecto, se desvela un polinucleotido, que comprende una secuencia que tiene al menos aproximadamente 20 %, por ejemplo, al menos aproximadamente 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 96 %, 97 %, 98 %, 99 % o 100 % de identidad con la SEQ ID NO:32 expuesta en la FIG. 59.

58. In one aspect, a polynucleotide encoding a precursor thioesterase is disclosed in which the precursor thioesterase comprises a protein having at least about 20%, for example, at least about 25%, 30%, 35%, 40%, 45 %, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with the sequence of a 'TesA of E.coli K12 bacteria . In one aspect, a polynucleotide is disclosed, which comprises a sequence having at least about 20%, for example, at least about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% , 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with SEQ ID NO: 32 set forth in FIG. 59.

En un aspecto, se desvela un vector que comprende un gen (o un polinucleotido) que codifica una tioesterasa mutante o uno de sus equivalentes naturales. Los vectores de acuerdo con la divulgacion pueden transformarse en celulas hospedadoras adecuadas para producir celulas hospedadoras recombinantes.

In one aspect, a vector is disclosed which comprises a gene (or a polynucleotide) that encodes a mutant thioesterase or one of its natural equivalents. The vectors according to the disclosure can be transformed into suitable host cells to produce recombinant host cells.

En un aspecto, se desvela una sonda que comprende un polinucleotido que tiene una longitud de aproximadamente 4 a aproximadamente 150 nucleotidos, que es sustancialmente identico a un fragmento correspondiente de la SEQ ID NO: 32 expuesta en la FIG. 59, en donde la sonda es util para detectar y/o identificar secuencias de polinucleotidos que codifican enzimas que tienen actividad tioesterasa. Segun la divulgacion, puede utilizarse una sonda para detectar y aislar posibles tioesterasas precursoras de fuentes desconocidas, para producir dichas tioesterasas precursoras o para las cuales no se conoce la secuencia de aminoacidos o de acidos nucleicos.

In one aspect, a probe comprising a polynucleotide having a length of about 4 to about 150 nucleotides is disclosed, which is substantially identical to a corresponding fragment of SEQ ID NO: 32 set forth in FIG. 59, wherein the probe is useful for detecting and / or identifying polynucleotide sequences encoding enzymes that have thioesterase activity. According to the disclosure, a probe to detect and isolate possible thioesterase precursors from unknown sources, to produce said thioesterase precursors or for which the sequence of amino acids or nucleic acids is not known.

En determinados aspectos, se desvela una celula hospedadora recombinante que comprende un polinucleotido que codifica una tioesterasa mutante o uno de sus equivalentes naturales. En un aspecto, para alterar o modificar las tioesterasas endogenas de la celula hospedadora pueden emplearse tecnicas conocidas de modificacion o alteracion genomica, efectuando una o mas de las mutaciones mencionadas anteriormente, de modo que al menos una de las tioesterasas endogenas mutantes tenga al menos una propiedad alterada. En otro aspecto, la celula hospedadora recombinante se genomanipula para incluir un plasmido que comprende un polinucleotido que codifica una tioesterasa mutante o uno de sus equivalentes naturales. En otro aspecto mas, la celula hospedadora recombinante expresa la tioesterasa despues de que el polinucleotido que codifica la tioesterasa se integre en el cromosoma de la celula hospedadora.

In certain aspects, a recombinant host cell is disclosed comprising a polynucleotide encoding a mutant thioesterase or one of its natural equivalents. In one aspect, known techniques of genomic modification or alteration can be used to alter or modify the endogenous thiosterases of the host cell, by performing one or more of the aforementioned mutations, so that at least one of the mutant endogenous thioesrases have at least one altered property. In another aspect, the recombinant host cell is genomanipulated to include a plasmid comprising a polynucleotide encoding a mutant thioesterase or one of its natural equivalents. In yet another aspect, the recombinant host cell expresses the thioesterase after the polynucleotide encoding the thioesterase is integrated into the chromosome of the host cell.

En un aspecto, la celula hospedadora recombinante de la divulgacion puede seleccionarse de cualquier celula que sea capaz de expresar una construccion genica recombinante, y puede seleccionarse de una celula microbiana, vegetal o animal. En un aspecto particular, el origen de la celula hospedadora es bacteriano, cianobacteriano, fungico, de levaduras, de algas, de ser humano o de mamifero. En un aspecto particular, la celula hospedadora se selecciona de cualquiera de las especies de bacterias grampositivas tales como Actinomycetes; Bacillaceae, incluyendo Bacillus alkalophilus, Bacillus subtilis, Bacillus licheniformis, Bacillus lentus, Bacillus brevis, Bacillus stearothermophilus, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus coagulans, Bacillus circulans, Bacillus lautus, Bacillus megaterium, B. thuringiensis; Brevibacteria sp., incluyendo Brevibacterium flavum, Brevibacterium lactofermentum, Brevibacterium ammoniagenes, Brevibacterium butanicum, Brevibacterium divaricatum, Brevibacterium healii, Brevibacterium ketoglutamicum, Brevibacterium ketosoreductum, Brevibacterium lactofermentum, Brevibacterium linens, Brevibacterium paraffinolyticum; Corynebacterium spp. tales como C. glutamicum y C. melassecola, Corynebacterium herculis, Corynebacterium lilium, Corynebactertium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium acetophilum, Corynebacterium ammoniagenes, Corynebacterium fujiokense, Corynebacterium nitrilophilus; o especies de bacterias acidolacticas incluyendo Lactococcus spp. tales como Lactococcus lactis; Lactobacillus spp. incluyendo Lactobacillus reuteri; Leuconostoc spp.; Pediococcus spp.; Serratia spp. tal como Serratia marcescens; especies del genero Streptomyces, tales como Streptomyces lividans, Streptomyces murinus, S. coelicolor y Streptococcus spp.. Como alternativa, cepas de una especie bacteriana gramnegativa perteneciente a la familia Enterobacteriaceae incluyendo E. coli, Cellulomonas spp.; o a la familia Pseudomonadaceae incluyendo Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas syringae y Burkholderia cepacia, Salmonella sp., Stenotrophomonas spp. y Stenotrophomonas maltophilia. tambien son utiles los microorganismos oleaginosos tales como Rhodococcus spp, Rhodococcus opacus, Ralstonia spp. y Acetinobacter spp.. Asimismo, las cepas de levaduras y de hongos filamentosos pueden ser celulas hospedadoras utiles, incluyendo Absidia spp.; Acremonium spp.; Agaricus spp.; Anaeromyces spp.; Aspergillus spp., incluyendo A. aculeatus, A. awamori, A. flavus, A. foetidus, A. fumaricus, A. fumigatus, A. nidulans, A. niger, A. oryzae, A. terreus; A. tubingensis y A. versicolor; Aeurobasidium spp.; Cephalosporum spp.; Chaetomium spp.; Coprinus spp.; Dactyllum spp.; Fusarium spp., incluyendo F. conglomerans, F. decemcellulare, F. javanicum, F. lini, F. oxysporum y F. solani; Gliocladium spp.; Kluyveromyces sp.; Hansenula sp.; Humicola spp., incluyendo H. insolens y H. lanuginosa; Hypocrea spp.; Mucor spp.; Neurospora spp., incluyendo N. crassa y N. sitophila; Neocallimastix spp.; Orpinomyces spp.; Penicillium spp.; Phanerochaete spp.; Phlebia spp.; Pichia sp.; Piromyces spp.; Rhizopus spp.; especies de Rizomucor tales como Rhizomucor miehei; Schizophyllum spp.; Schizosaccharomyces tal como, por ejemplo, la especie S. pombe; chytalidium sp., Sulpholobus sp., Thermoplasma sp., Thermomyces sp.; Trametes spp.; Trichoderma spp., incluyendo T. reesei, T. reesei (longibrachiatum) y T. viride; Yarrowinia sp.; y Zygorhynchus spp y en particular incluyendo precisamente las levaduras oleaginosas Phafia spp., Rhorosporidium toruloides Y4, Rhodotorula Glutinis y Candida 107.

In one aspect, the recombinant host cell of the disclosure can be selected from any cell that is capable of expressing a recombinant gene construct, and can be selected from a microbial, plant or animal cell. In a particular aspect, the origin of the host cell is bacterial, cyanobacterial, fungal, yeast, algae, human or mammalian. In a particular aspect, the host cell is selected from any of the gram-positive bacteria species such as Actinomycetes; Bacillaceae, including Bacillus alkalophilus, Bacillus subtilis, Bacillus licheniformis, Bacillus lentus, Bacillus brevis, Bacillus stearothermophilus, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus coagulans, Bacillus thursis, Bacillus circulants, Bacillus circulants Brevibacteria sp., Including Brevibacterium flavum, Brevibacterium lactofermentum, Brevibacterium ammoniagenes, Brevibacterium butanicum, Brevibacterium divaricatum, Brevibacterium healii, Brevibacterium ketoglutamicum, Brevibacterium ketosoreductum, Brevibacterium, Brevibacterium, Brevibacterium, Brevibacterium Corynebacterium spp. such as C. glutamicum and C. melassecola, Corynebacterium herculis, Corynebacterium lilium, Corynebactertium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium acetophilum, Corynebacterium ammoniagenes, Corynebacterium fujiokeium, Corynebacterium nitophnecterium; or species of acid lactic bacteria including Lactococcus spp. such as Lactococcus lactis; Lactobacillus spp. including Lactobacillus reuteri; Leuconostoc spp .; Pediococcus spp .; Serratia spp. just like Serratia marcescens; species of the genus Streptomyces, such as Streptomyces lividans, Streptomyces murinus, S. coelicolor and Streptococcus spp .. Alternatively, strains of a gram-negative bacterial species belonging to the Enterobacteriaceae family including E. coli, Cellulomonas spp .; or to the family Pseudomonadaceae including Pseudomonas aeruginosa, Pseudomonas alkalgenes, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas syringae and Burkholderia cepacia, Salmonella sp., Stenotrophomonas spp. and Stenotrophomonas maltophilia. Also useful are oleaginous microorganisms such as Rhodococcus spp, Rhodococcus opacus, Ralstonia spp. and Acetinobacter spp .. Also, strains of yeast and filamentous fungi may be useful host cells, including Absidia spp .; Acremonium spp .; Agaricus spp .; Anaeromyces spp .; Aspergillus spp., Including A. aculeatus, A. awamori, A. flavus, A. foetidus, A. fumaricus, A. fumigatus, A. nidulans, A. niger, A. oryzae, A. terreus; A. tubingensis and A. versicolor; Aeurobasidium spp .; Cephalosporum spp .; Chaetomium spp .; Coprinus spp .; Dactyllum spp .; Fusarium spp., Including F. conglomerans, F. decemcellulare, F. javanicum, F. lini, F. oxysporum and F. solani; Gliocladium spp .; Kluyveromyces sp .; Hansenula sp .; Humicola spp., Including H. insolens and H. lanuginosa; Hypocrea spp .; Mucor spp .; Neurospora spp., Including N. crassa and N. sitophila; Neocallimastix spp .; Orpinomyces spp .; Penicillium spp .; Phanerochaete spp .; Phlebia spp .; Pichia sp .; Piromyces spp .; Rhizopus spp .; Rizomucor species such as Rhizomucor miehei; Schizophyllum spp .; Schizosaccharomyces such as, for example, the S. pombe species ; chytalidium sp., Sulpholobus sp., Thermoplasma sp., Thermomyces sp .; Trametes spp .; Trichoderma spp., Including T. reesei, T. reesei ( longibrachiatum) and T. viride; Yarrowinia sp .; and Zygorhynchus spp and in particular including precisely the oil yeasts Phafia spp., Rhorosporidium toruloides Y4, Rhodotorula Glutinis and Candida 107.

En un aspecto, se desvela una celula hospedadora recombinante, que expresa o sobreexpresa un gen que codifica la tioesterasa mutante (o uno de sus equivalentes naturales), y que tambien expresa (o sobreexpresa) uno o mas genes que codifican una o mas enzimas que utilizan, como sustratos, productos de reaccion de la tioesterasa mutante (por ejemplo, acidos grasos, acil-CoA grasas, esteres grasos de acil-fosfato, aldehidos grasos, esteres grasos, o alcoholes grasos) o productos de reaccion de una o mas enzimas distintas que forman parte de una ruta metabolica, incluyendo productos de reaccion de la tioesterasa mutante (por ejemplo, acidos grasos) como precursores y/o sustratos.

In one aspect, a recombinant host cell is revealed, which expresses or overexpresses a gene that encodes the mutant thioesterase (or one of its natural equivalents), and that also expresses (or overexpresses) one or more genes encoding one or more enzymes that use, as substrates, reaction products of the mutant thioesterase (for example, fatty acids, fatty acyl-CoA, fatty esters of acyl phosphate, fatty aldehydes, fatty esters, or fatty alcohols) or reaction products of one or more enzymes distinct that are part of a metabolic pathway, including reaction products of the mutant thioesterase (eg, fatty acids) as precursors and / or substrates.

En un aspecto, se desvela una celula hospedadora recombinante, que expresa o sobreexpresa un gen que codifica una tioesterasa mutante (o uno de sus equivalentes naturales) y que tambien expresa (o sobreexpresa) uno o mas genes que codifican una o mas enzimas que reaccionan con un sustrato que es necesario como un precursor para una reaccion en una ruta biosintetica de acidos grasos. En un aspecto particular, la celula hospedadora recombinante incluye un gen que codifica la tioesterasa y un gen que codifica una enzima que reacciona con un sustrato que es necesario como un precursor para una reaccion en una ruta sintetica de acidos grasos, que comprende la sobreexpresion o modificacion de un gen seleccionado de pdh, pank, acef, fabH, fabD, fabG, acpP y/o fabF.

In one aspect, a recombinant host cell is revealed, which expresses or overexpresses a gene that encodes a mutant thioesterase (or one of its natural equivalents) and that also expresses (or overexpresses) one or more genes encoding one or more enzymes that react with a substrate that is necessary as a precursor for a reaction in a biosynthetic route of fatty acids. In a particular aspect, the recombinant host cell includes a gene that encodes thioesterase and a gene that encodes an enzyme that reacts with a substrate that is necessary as a precursor for a reaction in a synthetic fatty acid pathway, which comprises overexpression or modification of a gene selected from pdh, pank, acef, fabH, fabD, fabG, acpP and / or fabF.

En un aspecto , la ce lu la h o spe da do ra re co m b in a n te com p ren de un gen (o un p o lin u c le o tid o ) que cod ifica una t io e s te ra s a m u ta n te (o uno de sus e q u iva le n te s n a tu ra le s ) y ta m b ie n co m p re n d e la a te n u a c io n o de le c io n de un gen que reduce el flu jo de ca rbono , o un gen que com p ite po r sus tra to s , co fa c to res , o req u is itos de ene rg ia de n tro de una ru ta b ios in te tica de ac ido s g rasos. En un asp e c to pa rticu la r, el gen a te nua do com p ren de al m en os uno de los s ig u ie n te s ge ne s fadE, gpsA, IdhA, pflB, adhE, pta, poxB, ackA, ackB, pIsB y/o sfa.

In one aspect, the story of the gen- eration of a gene (or a small group) that encodes a guy is sam sam ta n te ( or one of its equations will show you) and it will also be the reason for the generation of a gene that reduces the flow of carbon, or a gene That it includes its traits, co-factors, or requirements of ene rg ia within one of the routes of a prior art of basic events. In an asp e r pa rrticu r, the gene has not yet com pled in at least one of the s f ss, gpsA, IdhA, pflB, adhE, pta, poxB , ackA, ackB, pIsB and / or sfa.

En un aspec to , una ce lu la h o sp e d a d o ra re co m b in a n te co m p re n d e un gen (o un p o lin u c le o tid o ) que cod ifica una t io e s te ra s a m u tan te (o uno de sus e q u iva le n te s n a tu ra les ) y un gen exogeno in tro d u c id o de m an e ra he te ro loga que cod ifica al m en os una en z im a de riva da de ac idos grasos. En d e te rm in a d o s aspectos, el gen exogeno o p o lin u c le o tid o cod ifica , po r e jem p lo , una a c il-C o A s in tasa , una e s te r s in tasa , una a lcoh o l a c iltra n s fe ra sa , una a lcoho l de sh id ro g e n a sa , una a c il-C o A reduc tasa , una a c il-C o A red uc tasa fo rm a d o ra de a lcoh o l graso, una ac ido ca rb o x ilico red uc tasa , una d e sca rb o x ila sa , una a ld e h id o red uc tasa , una a ce til tra n s fe ra s a de a lcoh o l graso , una en z im a co n d e n sa d o ra de ac ilo , una a m in o tra n s fe ra sa o una de sca rb on ila sa .

In one respect, a company with a gene (or a few languages) that encodes an uncle is a samou so te (o one of its equations will be your own) and an exogenous gene in tro duc id o de m an e ra has been a logo that codifies at least one of the fatty acids in the area of fatty acids . In all aspects, the exogenous gene opo lin uc le or tid or codifies, po re jem p lo, an ac il-C or A s in rate, one e s te rs in rate, one a lcoh olac iltra ns fe ra sa, a lcoho l of sh id ro gena sa, an ac il-C or A reduced rate, an ac il-C or A red uc fo rm for a lcoh ol fatty, an acid ca rb ox illico red uc rate, one of sca rb ox ila sa, one a ld eh id or red uc rate, one of the tradable services of a fatty alcohol, one in z co a co nden sa do ra of acyl, an am in or tra ns faith ra sa or one of sca rb on ila sa.

En un aspec to , la ce lu la h o sp e d a d o ra re c o m b in a n te c o m p re n d e un gen que co d ifica una t io e s te ra s a m u tan te (o uno de sus eq u iva le n te s n a tu ra les ) y al m en os dos genes e xo ge no s in tro d u c id o s de m an era he te ro loga que cod ifican en z im a s d e riva d a s de ac ido s g rasos. En d e te rm in a d o s aspec tos , los ge ne s o p o lin u c le o tid o s e xo g e n o s cod ifican , po r e jem p lo , una a c il-C o A s in tasa , una e s te r s in tasa , una a lcoh o l ac iltran s fe rasa , una a lcoh o l de sh id ro g e n a sa , una ac il-C o A reductasa , una a c il-C o A red uc tasa fo rm a d o ra de a lcoh o l graso, una ac ido ca rb ox ilico reduc tasa , una d e sca rb o x ila sa , una a ld e h id o reduc tasa , una a ce til tra n s fe ra s a de a lcoh o l graso , una e n z im a co n d e n sa d o ra de ac ilo , una a m in o tra n s fe ra sa o una de sca rb on ila sa .

In one respect, the city has been combined with a gene that encodes a guy who is so sam (or one of his eq u iva le n sna sna ra ra les) and at least in two genes that are not in tro ducts of m an era he r lo lo lo codify in z im asde riva das acidos sg rasos. In terms of specific aspects, the genes on the other side of the world codify, at least, an ac-C or A s in rate, one of them in rate, one to lcoh ol ac iltran s fe rasa, one to lcoh ol from sh id ro gena sa, an ac il-C or A reductase, an ac il-C or A red uc rate fo rm adora of a lcoh ol fatty, a Acid ca rb ox llico reduced rate, one of sca rb ox ila sa, one to ld eh id or reduced rate, one to other tradable company of a fatty alcohol, an enzyme known as of acyl, an am in or tra ns faith ra sa or one of sca rb on ila sa.

En un a sp ec to p re fe rido , un gen que cod ifica la t io e s te ra sa m u tan te (o uno de sus e q u iva le n te s n a tu ra le s ) y /o una e n z im a d e riva d a de a c id o s g ra sos , po r e jem p lo , una a c il-C o A s in tasa , una e s te r s in tasa , una a lcoh o l a c iltra n s fe ra sa , una a lcoh o l d e sh id ro g e n a sa , una a c il-C o A reduc tasa , una a c il-C o A red u c ta sa fo rm a d o ra de a lcoh o l graso , una ac ido c a rb o x ilico reduc tasa , una de sca rb ox ilasa , una a ld e h id o reduc tasa , una ace til tra n s fe ra s a de a lcoh o l graso, una e n z im a c o n d e n sa d o ra de ac ilo , una a lcoh o l ace tiltra n s fe ra sa , una a m in o tra n s fe ra sa , una tio e s te ra sa o una d e sca rb o x ila sa ad ic io n a l que se sob ree xp resa .

In an ecological context, a gene that encodes the uncle is te ra sa mu so te (or one of its equivalents) and / or an immersive enzyme It gives you a good idea, an ac c-C or A s in rate, one is in charge, one a lcoh olac iltra ns fe ra sa, one a lcoh olde sh id Genoa, an ac-C or A reduction rate, an ac-C or A red network for the purpose of a fatty alcohol, an acidic acid reduction rate, a tax rate ilasa, one year ago, a reduced rate, an acetyl transfer of a fatty alcohol, an enzyme of acyl acid, one year of an acetyl faith, one am or through faith, an uncle is tera sa or one of sca rb ox ila sa ad ic io nal that is under review.

En un aspecto , genes que cod ifican t io e s te ra sa s m u ta n te s (o sus eq u iva le n te s na tu ra les), enz im as de riva das de ac ido s g ra sos y/u o tros genes e xp re sa d o s de fo rm a reco m b in an te en una ce lu la h o sp e d a d o ra re co m b in an te , se m od ifica n para o p tim iz a r al m en os un codon para la exp res ion en la ce lu la h o sp e d a d o ra reco m b in an te .

In one aspect, genes that code for you are your son (or your eq u iva le n te s na tu ra les), enzymes of ac i riva das sg ra sos y / uo Three genes have been reported for a period of time in a city where they have been recorded in order to determine a codon for the Exp eration in the city of the city in Mexico.

En un aspec to , la ce lu la ho spe d a d o ra re co m b in a n te com p ren de al m en os un gen que cod ifica una t io e s te ra sa m u tan te (o uno de sus e q u iva le n tes n a tu ra les ) y un gen que cod ifica una a c il-C o A s in tasa . El gen de la ac il-C o A s in ta sa pued e s e r c u a lq u ie ra de los s ig u ie n te s genes, fadD, fadK, BH3103, yhfL, Pfl-4354, EAV15023, fadDI, fadD2, RPC_4074, fadDD35, fadDD22, faa3p, o el gen que cod ifica la p ro te ina Z P _ 01644857. O tro s e jem p los de genes de a c il-C o A s in ta sa inc luyen los genes fadDD35 de M. tuberculosis HR7Rv [N P _217021 ], yhfL de B. subtilis [N P _ 388908 ], fadD1 de P. aeruginosa PAO1 [N P _251989 ], el gen que cod ifica la p ro te ina Z P _ 01644857 de Stenotrophomonas maltophilia R551-3, o el gen faa3p de Saccharomyces cerevisiae [N P _012257 ].

In one respect, the company has given it a common idea that it is a gene that encodes an uncle that is very common (or one of its equations). n tes na tu ra les) and a gene that encodes an ac-C or A s in rate. The ac-C or A s in ta sa gene can be used in the same genes, fDD, fadK, BH3103, yhfL, Pfl-4354, EAV15023, fadDI, fadD2, RPC_4074 , fadDD35, fadDD22, faa3p, or the gene coding for the ZP _ 01644857 gene. Other genes of ac-C or A s in ta sa genes show the M. tuberculosis fadDD35 genes . HR7Rv [NP _217021], yhfL of B. subtilis [NP _ 388908], fadD1 of P. aeruginosa PAO1 [NP _251989], the gene that encodes the protein ZP _ 01644857 of Stenotrophomonas maltophilia R551-3, or faa3p gene of Saccharomyces cerevisiae [NP _012257].

En un aspecto , se de sve la una ce lu la h o sp e d a d o ra re co m b in a n te que com p ren de al m en os un gen o po lin uc leo tido que cod ifica una t io e s te ra s a m u tan te (o uno de sus e q u iva le n tes n a tu ra le s ) y un gen o p o lin uc leo tido que cod ifica una e s te r s in tasa , ta l com o un gen de e s te r s in ta sa ob te n id o de Acinetobacter spp., Alcanivorax borkumensis, Arabidopsis thaliana, Saccharomyces cerevisiae, Homo sapiens, Simmondsia chinensis, Mortierella alpina, Cryptococcus curvatus, Alcanivorax jadensis, Alcanivorax borkumensis, Acinetobacter sp. HO1-N, o Rhodococcus opacus. C om o e jem p los de genes de e s te r s in ta sa se inc luyen los genes wax/dgat, que cod ifican una es te r s in ta s a /a c il-C o A b ifu nc iona l: d ia c ilg lice ro l a c iltra n s fe ra sa de Simmondsia chinensis, Acinetobacter sp. cepa ADP1, Alcanivorax borkumensis, Pseudomonas aeruginosa, Fundibacter jadensis, Arabidopsis thaliana o Alkaligenes eutrophus. En un a sp e c to p re fe rido , el gen que cod ifica la e s te r s in ta sa es ta so b ree xp resa do .

In one aspect, it is the only one that has been accepted by a gene or group that codifies a guy who codifies a guy who is sam sam so te (or one of its equations le na tes na tu ra le s) and an opo lin uc le tido gene that encodes a te srs in rate, ta l com o a gene de e s te rs in ta sa ob te n id o of Acinetobacter spp., Alcanivorax borkumensis, Arabidopsis thaliana, Saccharomyces cerevisiae, Homo sapiens, Simmondsia chinensis, Mortierella alpina, Cryptococcus curvatus, Alcanivorax jadensis, Alcanivorax borkumebacter, Acinetorax borkumebacter. HO1-N, or Rhodococcus opacus. C om oe jem p those of the genes of this group include the wax / dgat genes , which encode a group in which the ac-C or A b ifu nc iona l: d ia c ilg lice ro lac iltra ns fe ra sa of Simmondsia chinensis, Acinetobacter sp. strain ADP1, Alcanivorax borkumensis, Pseudomonas aeruginosa, Fundibacter jadensis, Arabidopsis thaliana or Alkaligenes eutrophus. In an extended context, the gene coding for the te srs in ta sa is ta so b ree xp resa do.

En un aspecto , la ce lu la h o spe da do ra re co m b in a n te co m p re n d e al m en os un gen que cod ifica una en z im a b io s in te tica de a ld e h id o graso . Un gen que co d ifica una e n z im a b io s in te tica de a ld e h id o g ra so puede ser, po r e jem p lo , un gen de ac ido ca rb o x ilico red uc tasa (p o r e jem p lo , un gen car), que tie n e una se cu e n c ia de p o lin u c le o tid o s y/o un m o tivo de po lip ep tido s p ro p o rc io n a d o en los lis ta do s de las figu ras 32 y 33, o una v a ria n te de los m ism os. En a lg u n o s casos, el gen qu e c o d ifica una e n z im a b io s in te tica de a ld e h id o g ra so co d ifica uno o m as de los m o tivo s de a m in o a c id o s re p re se n ta d o s en la FIG . 33.

In one aspect, the house is known as a gene that encodes one of the most important techniques in the world. A gene that encodes an im ictive technique of a ld eh id d og ra so can be, po re jem p lo, a gene of acidic acid network uc rate (pore jem p lo, a car gene ), which has a signal of potential interest and / or a motive of positive interest in the lists of figures 32 and 33 , or a variety of the same. In some cases, the gene that echoes a non-technical enzyme in the United States is one or more of the reasons for am in or in all cases. It is noted in FIG. 33.

En un aspecto , la ce lu la h o spe da do ra reco m b in an te com p ren de al m en os un gen de p roducc ion de a lcoh o l graso. Los g e n e s de p ro du cc ion de a lco h o l g ra so inc luyen , po r e jem p lo , acr1. Los ge ne s de p ro du cc ion de a lcoh o l g ra so se de scrib en , po r e jem p lo , en las p u b lica c io n e s P C T n .°W O 2008 /119082 y W O 2007 /136762.

In one aspect, the house is known as a fatty gene for the production of at least one year. The genes of a lco holg ra so inc lu, p re jem p lo, acr1. The management of a lcoh olg ra so s is described in, po re jem p lo, in the PCT public publications No. WO 2008/119082 and WO 2007/136762.

En un aspec to , la ce lu la h o sp e d a d o ra re co m b in a n te co m p re n d e un gen q u e co d ifica una t io e s te ra s a m u ta n te (o uno de sus e q u iv a le n te s n a tu ra le s ) y un gen que co d ifica al m en os un gen p ro d u c to r de o le fina . El gen pued e s e r un gen p ro d u c to r de o le fin a te rm in a l o un gen p ro d u c to r de o le fina in te rna . C o m o e je m p lo s de ge ne s p ro d u c to re s de o le finas te rm in a le s , los de sc ritos en la P ub licac io n P C T No. 2009 /085278 , in c luye nd o el gen orj880, son ap rop iad os . C om o e je m p lo s de genes p ro d u c to re s de o le finas in te rnas, los de sc rito s en la pub licac ion P C T No. 2008 /147781 A 2 son ap rop iad os .

In one respect, the letter has been given to a gene that encodes an uncle (or one of its equities). sna tu ra le s) and a gene that codifies to me a gene that is fine. The gene can be a gene for the purpose of the end of a gene for the purpose of the end. How do I know how to get to know how to do it, those of the rites in the PCT P icacation No. 2009/085278, in c luye nd o el gene orj880, are ap rop iad os. As In the case of genes that are not only fine in the world, those of science in PCT publication No. 2008/147781 A 2 are supported.

En un aspec to , se p ro p o rc io n a una ce lu la h o spe da do ra re co m b in a n te que c o m p re n d e al m en os un gen o p o lin u c le o tid o q u e c o d ifica una t io e s te ra s a m u ta n te (o uno de sus e q u iv a le n te s na tu ra les ), y a l m en os uno de (a ) un gen o p o lin u c le o tid o que cod ifica una e n z im a de riva d a de ac ido s g ra sos y (b) un gen o po lin u c le o tid o que cod ifica una e n z im a a c il-C o A d e sh id ro g e n a sa que esta a tenuado . P re fe re n te m e n te , ese gen que c o d ifica una en z im a d e riva d a de ac ido g ra so que es ta a te n u a d o o d e le c io n a d o y que es e n d o g e n o a la ce lu la ho sp e d a d o ra , cod ifica , po r e jem p lo , una a c il-C o A s in tasa , una e s te r s in tasa , una a lcoh o l ac iltran s fe rasa , una a lcoh o l de sh id ro g e n a sa , una ac il-C o A reduc tasa , una ac ido ca rb o x ilico reduc tasa , una de sca rb on ila sa , una ace til tra n s fe ra s a de a lcoh o l graso , una d e sca rb o x ila sa de ac ido graso , o una a c il-C o A red uc tasa fo rm a d o ra de a lco h o l graso . En un aspecto , el gen a te n u a d o cod ifica una a c il-C o A s in ta sa o una e s te r s in tasa .

In one respect, it was possible to give a letter to the host that includes at least one gene opo lin uc le o tid oquecod ifica un un un un te es un te ra samu ta n te (or one of its teams in your country), and in one of (a) an opo lin uc le or tid gene that encodes a riva da enzyme In this way, you are and (b) a gene or potential that encodes an enzyme that is identical to the identity of a person who has been tended. P re fe re n tente, that gene that codifies one in z im rivative ac tive of g a so that it is not possible for you that is end or genoa ce lu la ho sp edado ra It codifies, po re jem p lo, an ac il-C or A s in rate, one e s te rs in rate, one a lcoh ol ac iltran s fe rasa, one a lcoh ol de sh id ro gena sa, an ac-C or A reduced rate, an acidic reduced rate, a scaled rate, an effective oil from a fatty company, a scaled rate of interest fatty acid, or an ac-C or A red uc fo rm adora rate of a lco hol fatty. In one aspect, the gene a te n u a d o encodes one a c il-C or A s in ta sa or an e s te r s in rate.

En un aspecto , se de sve la una ce lu la ho spe d a d o ra re co m b in a n te que exp resa , o p re fe re n te m e n te sob ree xp resa , una en z im a t io e s te ra s a en co n d ic io n e s que dan com o resu ltad o la s in tes is d irec ta de e s te re s g ra sos de ac il-A C P o ac il-C o A , ta le s com o es te re s m e tilico s de a c id o s g ra so s (F A M E , s ig las de l in g le s fatty acid methyl esters) y es te res e tilico s de ac ido s g ra sos (F A E E , fatty acid ethyl esters), m ed ia n te d icha tio e s te ra sa . En este aspec to , la tio e s te ra sa tra n s fo rm a d ire c ta m e n te la a c il-A C P o a c il-C o A en e s te r g ra so sin e x p re s a r n e ce sa ria m e n te n ing una e n z im a que sea una ac il C o A s in ta sa o una e s te r s in ta sa g rasa para p ro d u c ir es te res g rasos. No obs tan te , au nq ue la exp res ion o so b re e xp re s io n de una a c il-C o A s in ta sa o e s te r s in ta sa g rasa sea inne cesa ria , d ichas e n z im a s pueden se r d e sea b le s pa ra a u m e n ta r los re n d im ie n to s del p roducto . En este aspecto , la en z im a tio e s te ra s a puede se r cu a lq u ie ra de una t io e s te ra s a e n do ge na , una t io e s te ra s a e xp re sa d a de m an e ra he te ro loga , una t io e s te ra sa m utan te , o uno de sus e q u iva le n te s na tu ra les .

In one aspect, it is only one week after the expiration of the situation, op re faith re n te men te sob ree xp resa, one in z im at io es te ra sa in Co nd ic io nes that give com o resu ltad or the sis tes is the director of this re sg ra es de ac il-A CP or ac il-C or A, ta s s com or this re This is the case of fatty acid methyl esters (FAME, which is fatty acid methyl esters) and this is the case (FAEE, fatty acid ethyl esters), m ed ia n te d icha uncle es te ra sa. In this respect, the uncle is a member of the ad rire ad c ta men te la ac il il-A CP oac il-C or A en es s rg ra so without exp re sarne ce sa ria men te n ing an enzyme that is an ac C o A s in ta sa or an e s te rs in ta sa g rasa for this purpose. Not obs so te, even if the expiration or so b re e xp re s io n of an ac il-C or A s in ta sa or e s te in rs sa sa rasa be inde cesa ria, d These enzymes can be carried out in order to increase the revenue of the product. In this respect, the uncle is the only one who can know what an uncle is, a man is the best of all. ra lo lo ro loga, an uncle is te ra sa m utan te, or one of his team is his na s ra.

En un aspecto , la ce lu la h o sp e d a d o ra reco m b in an te tie n e un gen e n d o ge no que co d ifica una e n z im a ac il-C o A d e sh id ro g e n a sa que esta d e le c io n a d o o a tenuado .

In one aspect, the company has received an end-to-end gene that does not encode an enzyme of C-A or C of a type that is of le c io nadooa tenuado.

En un asp ec to , se d e sve la un m e to d o en d o n d e la ce lu la h o sp e d a d o ra re co m b in a n te se g u n la d ivu lg a c io n se cu ltiva en co n d ic io n e s que pe rm iten la exp res ion o so b re e xp re s io n de una o m as e n z im a s t io e s te ra sa s , que pueden se le cc io n a rse de tio e s te ra s a s endo ge nas , t io e s te ra sa s exp re sa d a s de m an e ra he te ro loga , tio e s te ra s a s m u tan tes (o sus eq u iva le n te s na tu ra les), o una com b in ac io n de estas t io e s te ra sa s . En un asp ec to pa rticu la r, la en z im a tio e s te ra s a que se e xp re sa o so b re e x p re s a pu e d e recu pe ra rse , y m as p re fe re n te m e n te , se pu rifica su s ta n c ia lm e n te , d e sp u e s de re co g e r y /o e fe c tu a r la lis is de la ce lu la ho spe da dora .

In an asp e ct, it is the one method in which the city has received a co-ordination in which the quantitative agreement is agreed upon For example, the reasons for the expiration of one or more enzymes are possible, which can be used as an uncle. , this is the reason for expiration of m an e ra he lo lo r loga, uncle es te ra sasmu so tees (or his eq u iva le n te s na tu ra les), or a com b in Here are these guys. In an ec e pa rrticu la r, the en z im a un un es te sa sa that e xp re sa o so b re exp re sa can be recovered, and more p re faith re n te ment te , its s ta nc ia lm en te, from sp u e s de re co gery / oe fe c tu ar la lis is de ce lu la ho spe da dora.

En un asp ec to , se d e sve la un m e to d o en d o n d e la ce lu la h o sp e d a d o ra re c o m b in a n te se g u n la d ivu lg a c io n se cu ltiva en co n d ic io n e s que pe rm iten la p ro du cc ion de d e riva d o s de ac ido s g rasos. En un a sp e c to p re fe rido , el de riva d o de ac ido g ra so puede recu pe ra rse , y m as p re fe ren te m en te , el d e riva d o de ac ido g ra so se pu rifica su s ta n c ia lm e n te . En un asp ec to pa rticu la rm e n te p re fe rido , la com p os ic ion de de riva dos de ac idos g ra sos se pu rifica s u s ta n c ia lm e n te de o tros co m p o n e n te s p ro d u c id o s d u ran te el cu ltivo m ed ia n te cen trifu ga c ion .

In an asp e ct, it is the one method in which the city has been re fl ered in the direction of the decision, which is quantitative in terms of conditions. That is the reason for the de riva dos de ac gone sgs. In a sp e c t e r r r fe r r, the riva do de ac gone ra ra so can be recovered, and more p r fe fe te te r in te, the riva do de ac a g gone so can be rifica su s ta nc ia lm in te. In an asp e r pa rrticu rm in te p re fe rido, the de riva two of acidic acids compares their rates in other countries as they constitute the sp duc id osdu ran the quantum m ed ia n te cen trifu ga tion.

En un aspecto , se de sve la un m e tod o para p ro d u c ir un de riva do de ac ido graso , que co m p re n d e c u ltiv a r una ce lu la h o sp e d a d o ra re co m b in a n te de la d ivu lg ac ion en co n d ic io n e s ad e cu a d a s para g a ra n tiz a r la exp res ion o s o b re e xp re s io n de una t io e s te ra s a m u ta n te (o uno de sus e q u iva le n te s na tu ra les), y re c u p e ra r el de riva d o de ac ido g ra so que se produce .

In one aspect, only one method of fatty acid is to be used, which includes a centenary of the cemetery in which d ivu lg ac ation in co adi cio ns ad e cu adas to guarantee the expiration osob re e xp re s io n of an un io es te ra samu ta n te (or one of its equ vat le n te s na tu rales), and recover that of riva do de ac ido ra so produced.

En un aspecto , se de sve la un m e tod o para p ro d u c ir e x tra ce lu la rm e n te un de riva do de ac ido g ra so in vitro, que co m p re n d e c u ltiv a r una ce lu la h o spe da do ra reco m b in an te en con d ic io n e s ad e cu a d a s para la exp res ion o so b re e xp re s io n de una en z im a tio e s te ra sa (in c luye nd o , po r e jem p lo , una t io e s te ra sa endo ge na , una t io e s te ra sa e xp re sa d a de m an e ra he te ro loga , una tio e s te ra s a m u tan te , o uno de sus e q u iva le n te s na tu ra les), re co g ie n d o y e fe c tu a n d o la lis is de las ce lu las , de ta l m an era que la en z im a tio e s te ra sa que se p roduce pueda recu pe ra rse y u tiliza rse para p ro d u c ir d e riva d o s de a c id o s g ra sos in vitro. En un asp ec to e jem p la r, la e n z im a tio e s te ra s a se pu rifica sus ta n c ia lm e n te . En o tro aspec to e jem p la r, la e n z im a tio e s te ra sa no se pu rifica de l lisado ce lu la r. La en z im a tio e s te ra s a pu rifica d a o el lisado c e lu la r que co m p re n d e d ich a enz im a , puede so m e te rse de sp u e s a su s tra to s de t io e s te ra s a a d e cu a d o s en c o n d ic io n e s que pe rm ita n la p ro d u cc io n e x tra c e lu la r de d e riva d o s de ac ido s g rasos. En la m a te ria se con oce n b ien te c n ic a s pa ra in tro d u c ir su s tra to s a enz im as. Un e jem p lo no lim itan te es a n a d ir el sus tra to (o sus tra to s ) en fo rm a de so luc ion a la so luc ion e n z im a tica o al lisado ce lu la r, y pe rm itir que la m ezc la se incube. O tro e jem p lo no lim itan te con s is te en in cu b a r el sus tra to (o sus tra to s ) y la so luc io n e n z im a tica o el lisado c e lu la r fija nd o el sus tra to (o su s tra to s ) o la enz im a a un m ed io so lido (p o r e jem p lo , perlas, res inas, p lacas, e tc .) y pa sa r la so luc io n e n z im a tica /lisa d o o el sus tra to (o sus tra to s), re sp ec tiva m en te , a tra v e s de l m ed io so lido a una ve lo c id a d que pe rm ita que se rea lice su fic ie n te co n ta c to e n tre el sus tra to (o sus tra to s ) y la enz im a .

In one aspect, it is only a matter of course to go outside the rm in a riva do de g ac ra ra so in vitro, which co r re ndecu ltiv ar a ce lu la ho spe da do ra reco mb in an te en con d ic io nes ad e cu adas for the exp resion or so b re e xp re s io n of an en z im a un un es es ra ra (in c luye n o o, po re jem p lo, an uncle is te ra sa end al ge na, an uncle is te ra sa e xp re sa da de m an e ra he lo ro loga, an uncle is te ra samu so te , or one of its teams is na na ra rales), re co g ie ndoye faith c tu and l la lis is de las ce lu las, so it was that in z im a uncle is te ra It is possible that it is possible to recover and use it so that it can be carried out in vitro. In an asp e to jem p la r, the enzyme is the reason that its rates are confirmed. In another aspect of the r, the enzyme is not purified from the lysate of the r. The en z im a uncle is te sa sa r rifica dao the lysate ce lu la r that c com d re nded to this im, can be told to me as a s te s t he un tio It is important to see in conditions that pe rm ita n p ro du cc io nex tra ce l la r de de riva dos de ac gone sos rasos. In the morning it is known that tea is also available in order to go to your system. A je lim p is not limited to your work (or your work) in order to solve an enzymatic solution or to the lysate ce, and it seems that the spirit is inculcated. O tro e jem p what is not limited to you in your bar, your treatment (or your dealings) and the economic solution or the lysate is fixed or fixed. treatment (or its treatment) or the enzyme is a solid medium (for example, pearls, resins, lacquers, etc.) and for the purpose of development. Ethical / Smooth Doo His Trafficking (or His Trafficking), Re sp ective, through Solid Soil at a time See what he wants to do Their fiction tells you about their treatment (or their treatment) and the enzyme.

En o tro aspec to , se d e sve la un m e tod o pa ra p ro d u c ir un d e riva d o de ac ido graso , que co m p re n d e c u ltiv a r una ce lu la h o sp e d a d o ra re co m b in a n te en co n d ic io n e s ad e cu a d a s para g a ra n tiz a r la exp res ion de una en z im a tio e s te ra sa (in c luye nd o , po r e jem p lo , una tio e s te ra sa endogena , una tio e s te ra sa e xp resa da de m an era he te ro loga , una t io e s te ra s a m u tan te , o uno de sus e q u iva le n te s na tu ra les), y re c u p e ra r el d e riva d o de ac ido g raso que se sec re ta o libe ra e x tra ce lu la rm e n te . P o r con s ig u ie n te , el p ro du c to de riva do de ac ido g raso se recu pe ra de, po r e jem p lo , el sobrenadante de un caldo de fermentacion en donde se cultiva la celula hospedadora.

In another respect, a method of fatty acidity is to be observed, which includes a celebration of the pasture of the country. In co nd ic io nes ad e cudas to ensure the expiration of one in z im a un te es sa ra (in c luye nd o, po re jem p lo, an uncle is te ra Endogenous, an uncle is te sa sa e xp resa da de m an era ro lo lo lo loga, an uncle es te ra samu tan te, or one of his equ siva le n s s na tu ra les), and re it covers the private sector that is separated or released abroad. For example, the source of the private sector is recovered from, potentially, the supernatant from a fermentation broth where the host cell is grown.

En un aspecto, se desvela un metodo para obtener extracelularmente una composicion de derivado de acido graso cultivando una celula hospedadora recombinante que se ha transformado con un polinucleotido que codifica una enzima tioesterasa (incluyendo, por ejemplo, una tioesterasa endogena, una tioesterasa heterologa, una tioesterasa mutante, o uno de sus equivalentes naturales), en condiciones que permitan la produccion de un derivado de acido graso, una parte mas grande o mas pequena de la cual se secreta o libera extracelularmente, y recuperar el derivado de acido graso que se produce. En un aspecto ejemplar, el derivado de acido graso se produce dentro de la celula, aunque la celula hospedadora libera una parte de este. Por consiguiente, el metodo comprende ademas la recogida de las celulas, su lisis y la recuperacion del derivado de acido graso.

In one aspect, a method for extracellularly obtaining a fatty acid derivative composition is disclosed by culturing a recombinant host cell that has been transformed with a polynucleotide encoding a thioesterase enzyme (including, for example, an endogenous thioesterase, a heterologous thioesterase, a mutant thioesterase, or one of its natural equivalents), under conditions that allow the production of a fatty acid derivative, a larger or smaller part of which is secreted or released extracellularly, and recover the fatty acid derivative that is produced . In an exemplary aspect, the fatty acid derivative is produced within the cell, although the host cell releases a portion thereof. Accordingly, the method further comprises the collection of the cells, their lysis and the recovery of the fatty acid derivative.

En un aspecto, se desvela un metodo para producir derivados de acidos grasos en donde una celula hospedadora recombinante que expresa, o preferentemente sobreexpresa, una enzima tioesterasa en condiciones que dan como resultado la sintesis de esteres grasos a partir de acil-ACP o acil-CoA mediante dicha tioesterasa, se cultiva en condiciones que permitan dicha produccion directa de esteres grasos.

In one aspect, a method for producing fatty acid derivatives is disclosed in which a recombinant host cell expressing, or preferably overexpressing, a thioesterase enzyme under conditions that result in the synthesis of fatty esters from acyl-ACP or acyl- CoA through said thioesterase, is grown under conditions that allow said direct production of fatty esters.

En un aspecto, se desvela un metodo para producir derivados de acidos grasos que comprende: modificar una o mas tioesterasas endogenas de la celula hospedadora utilizando tecnicas adecuadas de alteracion genomica de modo que las tioesterasas endogenas comprendan una o mas mutaciones y tengan una o mas propiedades alteradas, en comparacion con los precursores de las tioesterasas endogenas; y cultivar la celula hospedadora en condiciones adecuadas para que dicha celula hospedadora exprese o sobreexprese dichas tioesterasas mutantes; y recuperar los derivados de acidos grasos. En un aspecto ejemplar, el derivado de acido graso que se produce puede secretarse o liberarse extracelularmente, de modo que pueda recuperarse, por ejemplo, del sobrenadante del caldo de fermentacion en donde se cultiva la celula hospedadora.

In one aspect, a method for producing fatty acid derivatives is disclosed which comprises: modifying one or more endogenous thioesterase of the host cell using suitable genomic alteration techniques so that the endogenous thiosterases comprise one or more mutations and have one or more properties altered, compared to the precursors of endogenous thiosterases; and culturing the host cell under conditions suitable for said host cell to express or overexpress said mutant thiosterases; and recover fatty acid derivatives. In an exemplary aspect, the fatty acid derivative that is produced can be secreted or released extracellularly, so that it can be recovered, for example, from the fermentation broth supernatant where the host cell is grown.

En un aspecto, se desvela un metodo para producir derivados de acidos grasos que comprende: transformar la celula hospedadora con una secuencia de polinucleotidos que codifica una tioesterasa mutante (o uno de sus equivalentes naturales), de modo que la produccion de derivados de acidos grasos en la celula hospedadora se altera con respecto a una celula que no se ha transformado con el gen de tioesterasa mutante (o uno de sus equivalentes naturales).

In one aspect, a method for producing fatty acid derivatives is disclosed which comprises: transforming the host cell with a polynucleotide sequence encoding a mutant thioesterase (or one of its natural equivalents), so that the production of fatty acid derivatives in the host cell it is altered with respect to a cell that has not been transformed with the mutant thioesterase gene (or one of its natural equivalents).

En un aspecto, se desvela un metodo para producir derivados de acidos grasos que comprende: proporcionar una secuencia de polinucleotidos que comprende un gen que codifica una tioesterasa mutante (o uno de sus equivalentes naturales); transformar una celula hospedadora adecuada en condiciones en las que dicha secuencia de polinucleotidos se incorpore en dicho cromosoma de dicha celula y dicho gen pueda expresarse dentro de dicha celula hospedadora; cultivar la celula hospedadora transformada en condiciones adecuadas para que dicha celula hospedadora exprese dicho gen y produzca una proteina tioesterasa mutante (o uno de sus equivalentes naturales); y recuperar los derivados de acidos grasos.

In one aspect, a method for producing fatty acid derivatives is disclosed which comprises: providing a polynucleotide sequence comprising a gene encoding a mutant thioesterase (or one of its natural equivalents); transforming a suitable host cell under conditions in which said polynucleotide sequence is incorporated into said chromosome of said cell and said gene can be expressed within said host cell; culturing the transformed host cell under conditions suitable for said host cell to express said gene and produce a mutant thioesterase protein (or one of its natural equivalents); and recover fatty acid derivatives.

En cualquiera de los aspectos anteriores, los derivados de una longitud de cadena de carbono determinada pueden recuperarse a un mayor rendimiento proporcional, en comparacion con la produccion de dichos derivados de acidos grasos de la misma longitud de cadena de carbono en la misma celula hospedadora en ausencia de la tioesterasa mutante (o uno de sus equivalentes naturales). En un aspecto particular, los derivados de acidos grasos que se recuperan a un mayor o menor rendimiento comprenden una longitud de cadena primaria de Ce, C7, C8, C9, C10, C11, C12, C13, C14, C15, Cie, C17, C18, C19, C20, C2I, C22, C23, C24, C25, C26, C28, C29, C30, C31, C32, C33, C34, C35, C36, C37, C38 o C39 de cadena de acilo graso. Los derivados de acidos grasos que se recuperan que se recuperan a un mayor o menor rendimiento en la composicion, pueden seleccionarse de todos los tipos de derivados de acidos grasos, incluyendo, por ejemplo, hidrocarburos, acidos grasos, esteres grasos, aldehidos grasos, alcoholes grasos, olefinas terminales, olefinas internas, alcanos, dioles, aminas grasas, acidos dicarboxilicos o cetonas o combinaciones de los mismos.

In any of the above aspects, derivatives of a given carbon chain length can be recovered at a higher proportional yield, compared to the production of said fatty acid derivatives of the same carbon chain length in the same host cell in absence of mutant thioesterase (or one of its natural equivalents). In a particular aspect, the fatty acid derivatives that are recovered at a higher or lower yield comprise a primary chain length of Ce, C7, C8, C9, C10, C11, C12, C13, C14, C15, Cie, C17, C18, C19, C20, C2I, C22, C23, C24, C25, C26, C28, C29, C30, C31, C32, C33, C34, C35, C36, C37, C38 or C39 of fatty acyl chain. The fatty acid derivatives that are recovered that are recovered at a greater or lesser yield in the composition, can be selected from all types of fatty acid derivatives, including, for example, hydrocarbons, fatty acids, fatty esters, fatty aldehydes, alcohols fatty, terminal olefins, internal olefins, alkanes, diols, fatty amines, dicarboxylic acids or ketones or combinations thereof.

Como alternativa, en cualquiera de los aspectos anteriores, un derivado de acido graso particular puede producirse a un rendimiento proporcional o porcentual aumentado o disminuido en relacion con los otros derivados de acidos grasos, cuando se compara con el rendimiento proporcional o porcentual de ese derivado de acido graso en particular en la misma celula hospedadora en ausencia de la tioesterasa mutante (o uno de sus equivalentes naturales). En un aspecto particular, el derivado de acido graso que se produce a un rendimiento proporcional o porcentual aumentado es un ester graso. En otro aspecto, el derivado de acido graso que se produce a un rendimiento proporcional o porcentual reducido es un ester graso.

Alternatively, in any of the above aspects, a particular fatty acid derivative can be produced at an increased or decreased proportional or percentage yield relative to the other fatty acid derivatives, when compared to the proportional or percentage yield of that derivative of fatty acid in particular in the same host cell in the absence of the mutant thioesterase (or one of its natural equivalents). In a particular aspect, the fatty acid derivative that is produced at an increased proportional or percentage yield is a fatty ester. In another aspect, the fatty acid derivative that is produced at a reduced proportional or percentage yield is a fatty ester.

Como alternativa, en cualquiera de los aspectos anteriores, los derivados de acidos grasos pueden producirse a un mayor rendimiento, o a un mayor rendimiento proporcional de productos de cadena corta (por ejemplo, C8, C9, C10, C11, C12, C13 o C14). A la inversa, en cualquiera de los aspectos anteriores, los derivados de acidos grasos pueden producirse a un rendimiento reducido, o a un rendimiento proporcional reducido de productos de cadena corta (por ejemplo, C8, C9, C10, C11, C12, C13 o C14).

Alternatively, in any of the above aspects, fatty acid derivatives can be produced at a higher yield, or at a higher proportional yield of short chain products (eg, C8, C9, C10, C11, C12, C13 or C14) . Conversely, in any of the above aspects, fatty acid derivatives can be produced at a reduced yield, or at a reduced proportional yield of short chain products (e.g., C8, C9, C10, C11, C12, C13 or C14 ).

En un aspecto, se desvela un metodo de produccion de derivados de acidos grasos, en donde el rendimiento de derivados de acidos grasos producidos por el metodo de la divulgacion es de al menos aproximadamente 0,001 g de producto derivado de acido graso/g de fuente de carbono, por ejemplo, al menos aproximadamente 0,01 g de producto derivado de acido graso/g de fuente de carbono, aproximadamente 0,1 g de producto derivado de acido graso/g de fuente de carbono, aproximadamente 0,2 g de producto derivado de acido graso/g de fuente de carbono, aproximadamente 0,3 g de producto derivado de acido graso/g de fuente de carbono, aproximadamente 0,4 g de producto derivado de acido graso/g de fuente de carbono o de aproximadamente 0,45 g de producto derivado de acido graso/g de fuente de carbono.

In one aspect, a method of producing fatty acid derivatives is disclosed, wherein the yield of fatty acid derivatives produced by the method of disclosure is at least about 0.001 g of product derived from fatty acid / g from carbon source, for example, at least about 0.01 g of product derived from fatty acid / g of carbon source, approximately 0.1 g of product derived from fatty acid / g from carbon source, approximately 0.2 g of product derived from fatty acid / g from carbon source, approximately 0.3 g of product derived from acid fat / g of carbon source, about 0.4 g of product derived from fatty acid / g of carbon source or about 0.45 g of product derived from fatty acid / g of carbon source.

En un aspecto, se desvela un metodo de produccion de derivados de acidos grasos, en donde el metodo da como resultado un titulo de al menos aproximadamente 0,5 g/l, por ejemplo, al menos aproximadamente 1 g/l, 2 g/l, 5 g/l, 10 g/l, 20 g/l, 30 g/l, 40 g/l, 50 g/l, 75 g/l, 100 g/l, 150 g/l o 200 g/l.

In one aspect, a method of producing fatty acid derivatives is disclosed, wherein the method results in a titer of at least about 0.5 g / l, for example, at least about 1 g / l, 2 g / l, 5 g / l, 10 g / l, 20 g / l, 30 g / l, 40 g / l, 50 g / l, 75 g / l, 100 g / l, 150 g / l or 200 g / l .

En un aspecto, se desvela un metodo de produccion de derivados de acidos grasos en donde la productividad del metodo es tal que se producen al menos aproximadamente 0,1 g/l.h, por ejemplo, al menos aproximadamente 0,5 g/l.h, 1 g/l.h, 2 g/l.h, 3 g/l.h, 4 g/l.h, 5 g/l.h, 6 g/l.h, 7 g/l.h u 8 g/l.h.

In one aspect, a method of producing fatty acid derivatives is disclosed wherein the productivity of the method is such that at least about 0.1 g / lh is produced, for example, at least about 0.5 g / lh, 1 g / lh, 2 g / lh, 3 g / lh, 4 g / lh, 5 g / lh, 6 g / lh, 7 g / lh or 8 g / lh

En un aspecto, se desvelan composiciones de derivados de acidos grasos que producen las celulas hospedadoras de la divulgacion. Dichas composiciones pueden comprender hidrocarburos, esteres, alcoholes, cetonas, aldehidos, acidos grasos, acidos dicarboxilicos, olefinas internas, olefinas terminales y/o combinaciones de los mismos. Dichas composiciones son utiles en aplicaciones en la industria quimica, por ejemplo, en la produccion de tensioactivos y detergentes, o como un biocombustible y un sustituto del petroleo, gasoil para calefaccion, queroseno, diesel, combustible de aviacion o gasolina.

In one aspect, fatty acid derivative compositions that produce the host cells of the disclosure are disclosed. Such compositions may comprise hydrocarbons, esters, alcohols, ketones, aldehydes, fatty acids, dicarboxylic acids, internal olefins, terminal olefins and / or combinations thereof. Such compositions are useful in applications in the chemical industry, for example, in the production of surfactants and detergents, or as a biofuel and a substitute for petroleum, heating oil, kerosene, diesel, aviation fuel or gasoline.

En un aspecto, se desvelan composiciones de derivados de acidos grasos que comprenden una cantidad menor o igual a aproximadamente 50 ppm de arsenico, aproximadamente 30 ppm, aproximadamente 25 ppm, o entre aproximadamente 10 y aproximadamente 50 ppm de arsenico; menor o igual a aproximadamente 200 ppm de calcio, aproximadamente 150 ppm de calcio, aproximadamente 119 ppm de calcio o entre aproximadamente 50 y aproximadamente 200 ppm de calcio; menor o igual a aproximadamente 200 ppm de cloro, aproximadamente 150 ppm de cloro, aproximadamente 119 ppm de cloro o entre aproximadamente 50 y aproximadamente 200 ppm de cloro; menor o igual a aproximadamente 50 ppm de cobre, aproximadamente 30 ppm de cobre, aproximadamente 23 ppm de cobre, o entre aproximadamente 10 y aproximadamente 50 ppm de cobre; menor o igual a aproximadamente 300 ppm de hierro, aproximadamente 200 ppm de hierro, aproximadamente 136 ppm de hierro, o entre aproximadamente 50 y aproximadamente 250 ppm de hierro; menor o igual a aproximadamente 50 ppm de plomo, aproximadamente 30 ppm de plomo, aproximadamente 25 ppm de plomo, o entre aproximadamente 10 y aproximadamente 50 ppm de plomo; menor o igual a aproximadamente 50 ppm de manganeso, aproximadamente 30 ppm de manganeso, aproximadamente 23 ppm de manganeso, o entre aproximadamente 10 y aproximadamente 50 ppm de manganeso; menor o igual a aproximadamente 50 ppm de magnesio, aproximadamente 30 ppm de magnesio, aproximadamente 23 ppm de magnesio, o entre aproximadamente 10 y aproximadamente 50 ppm de magnesio; menor o igual a aproximadamente 0,5 ppm de mercurio, aproximadamente 0,1 ppm de mercurio, aproximadamente 0,06 ppm de mercurio o entre aproximadamente 0,01 y aproximadamente 0,2 ppm de mercurio; menor o igual a aproximadamente 50 ppm de molibdeno, aproximadamente 30 ppm de molibdeno, aproximadamente 23 ppm de molibdeno o entre aproximadamente 10 y aproximadamente 50 ppm de molibdeno; menor o igual a aproximadamente 2 % de nitrogeno; aproximadamente 1 % de nitrogeno, aproximadamente 0,5 % de nitrogeno, o entre aproximadamente 0,1-1 % de nitrogeno; menor o igual a aproximadamente 200 ppm de potasio, aproximadamente 150 ppm de potasio, aproximadamente 103 ppm de potasio, o entre aproximadamente 50 y aproximadamente 200 ppm de potasio; menor o igual a aproximadamente 300 ppm de sodio, 200 ppm de sodio, aproximadamente 140 ppm de sodio, o entre aproximadamente 50 y aproximadamente 300 ppm de sodio; menor o igual a aproximadamente 1 ppm de azufre, menor o igual a aproximadamente 1 % de azufre, aproximadamente 0,14% de azufre, o entre aproximadamente 0,05 y aproximadamente 0,3% de azufre; menor o igual a aproximadamente 50 ppm de cinc, aproximadamente 30 ppm de cinc, aproximadamente 23 ppm de cinc, o entre aproximadamente 10 y aproximadamente 50 ppm de cinc; o menor o igual a aproximadamente 700 ppm de fosforo, aproximadamente 500 ppm de fosforo, aproximadamente 350 ppm de fosforo, o entre aproximadamente 100 y aproximadamente 700 ppm de fosforo.

In one aspect, fatty acid derivative compositions comprising an amount less than or equal to about 50 ppm of arsenic, about 30 ppm, about 25 ppm, or between about 10 and about 50 ppm of arsenic are disclosed; less than or equal to about 200 ppm of calcium, about 150 ppm of calcium, about 119 ppm of calcium or between about 50 and about 200 ppm of calcium; less than or equal to about 200 ppm of chlorine, about 150 ppm of chlorine, about 119 ppm of chlorine or between about 50 and about 200 ppm of chlorine; less than or equal to approximately 50 ppm copper, approximately 30 ppm copper, approximately 23 ppm copper, or between approximately 10 and approximately 50 ppm copper; less than or equal to about 300 ppm of iron, about 200 ppm of iron, about 136 ppm of iron, or between about 50 and about 250 ppm of iron; less than or equal to about 50 ppm of lead, about 30 ppm of lead, about 25 ppm of lead, or between about 10 and about 50 ppm of lead; less than or equal to about 50 ppm of manganese, about 30 ppm of manganese, about 23 ppm of manganese, or between about 10 and about 50 ppm of manganese; less than or equal to about 50 ppm of magnesium, about 30 ppm of magnesium, about 23 ppm of magnesium, or between about 10 and about 50 ppm of magnesium; less than or equal to about 0.5 ppm of mercury, about 0.1 ppm of mercury, about 0.06 ppm of mercury or between about 0.01 and about 0.2 ppm of mercury; less than or equal to about 50 ppm of molybdenum, about 30 ppm of molybdenum, about 23 ppm of molybdenum or between about 10 and about 50 ppm of molybdenum; less than or equal to about 2% nitrogen; about 1% nitrogen, about 0.5% nitrogen, or between about 0.1-1% nitrogen; less than or equal to about 200 ppm of potassium, about 150 ppm of potassium, about 103 ppm of potassium, or between about 50 and about 200 ppm of potassium; less than or equal to about 300 ppm of sodium, 200 ppm of sodium, about 140 ppm of sodium, or between about 50 and about 300 ppm of sodium; less than or equal to about 1 ppm sulfur, less than or equal to about 1% sulfur, about 0.14% sulfur, or between about 0.05 and about 0.3% sulfur; less than or equal to about 50 ppm of zinc, about 30 ppm of zinc, about 23 ppm of zinc, or between about 10 and about 50 ppm of zinc; or less than or equal to approximately 700 ppm phosphorus, approximately 500 ppm phosphorus, approximately 350 ppm phosphorus, or between approximately 100 and approximately 700 ppm phosphorus.

En un aspecto, se desvelan derivados de acidos grasos que tienen fracciones de carbono moderno de aproximadamente 1,003 a aproximadamente 1,5.

In one aspect, fatty acid derivatives having modern carbon fractions of about 1,003 to about 1.5 are disclosed.

En un aspecto, se desvela una composicion de derivado de acido graso en donde la composicion incluye constituyentes que comprenden un grupo acilo que tiene un doble enlace en la posicion 7 en la cadena de carbono (entre el numero de carbono 7 en la cadena de carbono y el numero de carbono 8 en la cadena de carbono) desde su extremo reducido.

In one aspect, a fatty acid derivative composition is disclosed wherein the composition includes constituents comprising an acyl group having a double bond at position 7 in the carbon chain (between carbon number 7 in the carbon chain and the number of carbon 8 in the carbon chain) from its reduced end.

En un aspecto particular, la composicion de derivados de acidos grasos comprende esteres grasos C5-C25 (es decir, una longitud de cadena de carbono de 5 a 25 carbonos), acidos grasos C5-C25, aldehidos grasos C5-C25, alcoholes grasos C5-C25; o esteres acidos C10-C20 (es decir, una longitud de cadena de carbono de 10 a 20 carbonos),acidos grasos C10-C20, aldehidos grasos C10-C20, alcoholes grasos C10-C20; o esteres grasos C12-C18 (es decir, una longitud de ca d e n a de c a rb o n o de 12 a 18 ca rb o n o s ),a c id o s g ra so s C ¹² -C ¹⁸ , a ld e h id o s g ra so s C ¹²-C ¹⁸ , a lco h o le s g ra so s C ¹² -C ¹⁸.

In a particular aspect, the composition of fatty acid derivatives comprises C5-C25 fatty esters (ie, a carbon chain length of 5 to 25 carbons), C5-C25 fatty acids, C5-C25 fatty aldehydes, C5 fatty alcohols -C25; or C10-C20 acid esters (ie, a carbon chain length of 10 to 20 carbons), C10-C20 fatty acids, C10-C20 fatty aldehydes, C10-C20 fatty alcohols; or C12-C18 fatty esters (i.e. a length from Cana de Ca Rb Ono from 12 to 18 Ca rb onos), a c id osg ra so s C ¹² -C ¹⁸ , a ld eh id osg ra so s C ¹² -C ¹⁸ , a lco ho le sg ra so s C ¹² -C ¹⁸ .

En un a sp e c to pa rticu la r, los de riva dos de ac ido s g ra sos de la d ivu lg ac ion com p re n d e n de riva dos de ac ido s g ra sos de ca d e n a linea l, d e riva d o s de ac ido s g ra sos de ca d e n a ra m ifica d a y /o g ru p o s c ic licos . En un a sp e c to pa rticu la r, los d e riva d o s de ac ido s g ra sos son in sa tu ra d o s (p o r e jem p lo , m o n o in sa tu ra d o s ) o sa tu rados .

In a sp e c e pa rrticu la r, those of riva dos de ac gone sg ra are from the d ivu lg ac ion com p rnden de riva dos de acc sg ra sos de ca dena line l, of riva dos In this way, you are from the day of the day. In a sp e c to pa rticu la r, the d d riva d o s s g ra sos are in sa tu ra d o s (p o r e jem p lo, m o n o in sa your ra d o s) or sa tu rados.

En un asp ec to , la c o m p o s ic io n de d e riv a d o s de a c id o s g ra s o s c o m p re n d e un e s te r g ra so q u e se p ro d u ce a p a rtir de un a lcoh o l y una ac il-C oA , en do nde el a lcoh o l t ie n e una lo ng itud de al m en os a p ro x im a d a m e n te 1, po r e jem p lo , de al m en os a p ro x im a d a m e n te 2, a p ro x im a d a m e n te 3, a p ro x im a d a m e n te 4, a p ro x im a d a m e n te 5, a p ro x im a d a m e n te 6, a p ro x im a d a m e n te 7, a p ro x im a d a m e n te 8, a p ro x im a d a m e n te 10, a p ro x im a d a m e n te 12, a p ro x im a d a m e n te 14, a p ro x im a d a m e n te 16 o a p ro x im a d a m e n te 18 ca rb o n o s y la a c il-C o A tie n e una lo ng itud de al m enos a p ro x im a d a m e n te 2, po r e jem p lo , de al m en os a p ro x im a d a m e n te 4, a p ro x im a d a m e n te 6, a p ro x im a d a m e n te 8, a p ro x im a d a m e n te 10, a p ro x im a d a m e n te 12, a p ro x im a d a m e n te 14, a p ro x im a d a m e n te 16, a p ro x im a d a m e n te 18, a p ro x im a d a m e n te 20, a p ro x im a d a m e n te 22, a p ro x im a d a m e n te 24 o a p ro x im a d a m e n te 26 ca rbonos . En a lgunos asp ec tos , el a lco h o l y la ac il-C oA , a p a rtir de los cua le s se p ro du cen los es te re s g rasos, va ria n en a p ro x im a d a m e n te 2, a p ro x im a d a m e n te 4, a p ro x im a d a m e n te 6, a p ro x im a d a m e n te 8, a p ro x im a d a m e n te 10, a p ro x im a d a m e n te 12 o a p ro x im a d a m e n te 14 a to m o s de carbono .

In an asp e ct, the composition of the income of the company is supported by a group so that it is possible to appear from one year ago and one year ago. , where el lcoh olt ie ne un lo ng itud de al m in os ap ro x im adamen te 1, po re jem p lo, of al m in os ap ro x im adamen te 2, ap ro x im adamen te 3, ap ro x im adamen te 4, ap ro x im adamen te 5, ap ro x im adamen te 6, ap ro x im adamen te 7, ap ro x im adamen te 8, ap ro x im adamen te 10, ap ro x im adamen te 12, ap ro x im adamen te 14, ap ro x im adamen te 16 oap ro x im adamen te 18 ca rb onosy la ac il-C o A ne ne lo ng itud al m enos ap ro x im adamen te 2, po re jem p lo, from al m en os ap ro x im adamen te 4, ap ro x im adamen te 6, ap ro x im adamen te 8, ap ro x im adamen te 10, ap ro x im adamen te 12, ap ro x im adamen te 14, apr o x im a d a m e n te 16, a p ro x im a d a m e n te 18, a p ro x im a d a m e n te 20, a p ro x im a d a m e n te 22, a p ro x im a d a m e n te 24 o a p ro x im a d a m e n te 26 char. In some cases, the Holy Spirit, the ac-c oA, away from those who are behind them, will go on ap r x im adamen te 2, ap ro x im adamen te 4, ap ro x im adamen te 6, ap ro x im adamen te 8, ap ro x im adamen te 10, ap ro x im adamen te 12 oap ro x im adamen te 14 to carbon .

En o tro aspec to , la co m p o s ic io n de d e riva d o s de ac ido s g ra sos c o m p re n d e un e s te r g ra so que se p ro du ce a pa rtir de un a lcoh o l y una ac il-A C P , en dond e el a lcoh o l t ie n e una lo ng itud de al m en os a p ro x im a d a m e n te 1, po r e jem p lo , al m en os de a p ro x im a d a m e n te 2, a p ro x im a d a m e n te 3, a p ro x im a d a m e n te 4, a p ro x im a d a m e n te 5, a p ro x im a d a m e n te 6, a p ro x im a d a m e n te 7, a p ro x im a d a m e n te 8, a p ro x im a d a m e n te 10, a p ro x im a d a m e n te 12, a p ro x im a d a m e n te 14, a p ro x im a d a m e n te 16 o a p ro x im a d a m e n te 18 ca rb on os y la ac il-A C P tie n e una long itud de a p ro x im a d a m e n te 2, por e jem p lo , a p ro x im a d a m e n te 4, a p ro x im a d a m e n te 6, a p ro x im a d a m e n te 8, a p ro x im a d a m e n te 10, a p ro x im a d a m e n te 12, a p ro x im a d a m e n te 14, a p ro x im a d a m e n te 16, a p ro x im a d a m e n te 18, a p ro x im a d a m e n te 20, a p ro x im a d a m e n te 22, a p ro x im a d a m e n te 24 o a p ro x im a d a m e n te 26 ca rb on os . En a lg u n o s asp ec tos , el a lcoh o l y la ac il-A C P , a p a rtir de los cua le s se p roduce el e s te r graso , va ria n en a p ro x im a d a m e n te 2, a p ro x im a d a m e n te 4, a p ro x im a d a m e n te 6, a p ro x im a d a m e n te 8, a p ro x im a d a m e n te 10, a p ro x im a d a m e n te 12 o a p ro x im a d a m e n te 14 a to m o s de carbono.

In other respects, the de riva dos de riva dos de ac s ra sos te so sos sos sos sos sos sos sos so so so so so so so so que que que que que que que que que que que que To CP, in which he has chosen a lot of money in the first year, he said it, at least in the second year, two times x im adamen te 3, ap ro x im adamen te 4, ap ro x im adamen te 5, ap ro x im adamen te 6, ap ro x im adamen te 7, ap ro x im adamen te 8, ap ro x im adamen te 10, ap ro x im adamen te 12, ap ro x im adamen te 14, ap ro x im adamen te 16 oap ro x im adamen te 18 ca rb on os and ac il-A CP has a long itud from ap ro x im adamen te 2, by e jem p lo, ap ro x im adamen te 4, ap ro x im adamen te 6, ap ro x im adamen te 8, ap ro x im adamen te 10, ap ro x im adamen te 12, ap ro x im adamen te 14, ap ro x im adamen te 16, ap ro xi m a d a m e n te 18, a p ro x im a d a m e n te 20, a p ro x im a d a m e n te 22, a p ro x im a d a m e n te 24 o a p ro x im a d a m e n te 26 ca rb on os. In some cases, the former and the Ac-CP, to turn off the fatty acids, will be available in April 2, April 2 x im adamen te 4, ap ro x im adamen te 6, ap ro x im adamen te 8, ap ro x im adamen te 10, ap ro x im adamen te 12 oap ro x im adamen te 14 to carbon.

En un aspecto , la com p o s ic io n de de riva dos de ac idos g ra sos co m p re n d e una m ezc la de de riva dos que inc luyen ac ido s g ra so s lib res. En un aspec to , el p o rce n ta je en peso de ac ido s g ra so s lib res es de al m en os a p ro x im a d a m e n te 0 ,5 %, po r e jem p lo , de al m en os a p ro x im a d a m e n te 1 %, a p ro x im a d a m e n te 2 %, a p ro x im a d a m e n te 3 %, a p ro x im a d a m e n te 4 %, a p ro x im a d a m e n te 5 %, a p ro x im a d a m e n te 6 %, a p ro x im a d a m e n te 7 %, a p ro x im a d a m e n te 8 % , a p ro x im a d a m e n te 9 % , a p ro x im a d a m e n te 10 % , a p ro x im a d a m e n te 15 % , a p ro x im a d a m e n te 20 % o de a p ro x im a d a m e n te 25 %. En un asp ec to de te rm in ado , el po rcen ta je en peso de e s te re s g ra sos p ro du c ido s es de al m en os a p ro x im a d a m e n te 50 %, po r e jem p lo , de al m en os a p ro x im a d a m e n te 55 %, a p ro x im a d a m e n te 60 %, a p ro x im a d a m e n te 65 %, a p ro x im a d a m e n te 70 %, a p ro x im a d a m e n te 75 %, a p ro x im a d a m e n te 80 %, a p ro x im a d a m e n te 85 % o de a p ro x im a d a m e n te 90 %. En un a sp ec to a d ic io na l, la re lac ion en peso de de riva dos de a c id o s g ra so s d is tin to s de ac ido s g ra so s lib res con re sp e c to a a c id o s g ra so s lib res, es m a yo r que a p ro x im a d a m e n te 90 :1 , po r e jem p lo , m a yo r que a p ro x im a d a m e n te 80:1, a p ro x im a d a m e n te 50:1, a p ro x im a d a m e n te 20:1, a p ro x im a d a m e n te 10:1, a p ro x im a d a m e n te 9:1, a p ro x im a d a m e n te 8:1, a p ro x im a d a m e n te 7:1, a p ro x im a d a m e n te 5:1, a p ro x im a d a m e n te 2:1 o a p ro x im a d a m e n te 1:1.

In one aspect, the comparison of two types of common acids is also one of the two that includes two acids. In one respect, the power by weight of the accrued sg ra so s lib res is at least 0.5%, by weight, from at least in years. ap ro x im adamen te 1%, ap ro x im adamen te 2%, ap ro x im adamen te 3%, ap ro x im adamen te 4%, ap ro x im adamen te 5%, ap ro x im adamen 6%, ap ro x im adamen te 7%, ap ro x im adamen te 8%, ap ro x im adamen te 9%, ap ro x im adamen te 10%, ap ro x im adamen te 15%, ap ro x im adamen te 20% or ap ro x im adamen te 25%. In an asp e te rm in ado, the po rce ta je in weight of this re sg ra es s p ro du c gone s is at the same time ap ro x im adamen te 50%, po re jem p lo, from at m in os ap ro x im adamen te 55%, ap ro x im adamen te 60%, ap ro x im adamen te 65%, ap ro x im adamen te 70%, ap ro x im adamen te 75%, ap ro x im adamen te 80%, ap ro x im adamen te 85% or ap ro x im adamen te 90%. In an ad sp e to ad ic io na l, the weight re rtion of de riva dos de a c id osg ra so sd is tin to s de ac a sg ra so s lib res with re sp ec to a a c id osg ra so s lib res, is ma me r that ap ro x im adamen te 90: 1, po re jem p lo, ma yo r that ap ro x im adamen te 80: 1, ap ro x im adamen te 50: 1, ap ro x im adamen te 20: 1, ap ro x im adamen te 10: 1, ap ro x im adamen te 9: 1, ap ro x im adamen te 8: 1, ap ro x im adamen te 7: 1, ap ro x im adamen te 5: 1, ap ro x im adamen te 2: 1 oap ro x im adamen te 1: 1.

En un aspec to , la co m p o s ic io n de de riva dos de ac ido s g ra sos com p ren de una m ezc la de de riva dos que inc luye ac ido s g ra sos lib res. En un aspec to , el p o rce n ta je en peso de ac ido s g ra sos lib res es de al m en os a p ro x im a d a m e n te 50 %, por e jem p lo , de al m en os a p ro x im a d a m e n te 55 %, a p ro x im a d a m e n te 60 %, a p ro x im a d a m e n te 65 %, a p ro x im a d a m e n te 70 %, a p ro x im a d a m e n te 75 %, a p ro x im a d a m e n te 80 %, a p ro x im a d a m e n te 85 % o de a p ro x im a d a m e n te 90 %. En un asp e c to d e te rm in ado , el po rcen ta je en peso de e s te r g ra so p ro du c ido es de al m enos a p ro x im a d a m e n te 0 ,5 % , po r e jem p lo , de al m en os a p ro x im a d a m e n te 1 % , a p ro x im a d a m e n te 2 % , a p ro x im a d a m e n te 3 %, a p ro x im a d a m e n te 4 %, a p ro x im a d a m e n te 5 %, a p ro x im a d a m e n te 6 %, a p ro x im a d a m e n te 7 %, a p ro x im a d a m e n te 8 %, a p ro x im a d a m e n te 9 %, a p ro x im a d a m e n te 10 %, a p ro x im a d a m e n te 15 %, a p ro x im a d a m e n te 20 %, a p ro x im a d a m e n te 25 %, a p ro x im a d a m e n te 30 %, a p ro x im a d a m e n te 35 %, a p ro x im a d a m e n te 40 %, a p ro x im a d a m e n te 45 % o de a p ro x im a d a m e n te 50 %. En un asp ec to ad ic iona l, la re lac ion en peso de de riva dos de ac ido s g ra sos p ro du c ido s d is tin to s de ac idos g ra sos lib res con respec to a ac idos g rasos lib res es m e n o r que a p ro x im a d a m e n te 60 :1 , po r e jem p lo , m e n o r que a p ro x im a d a m e n te 50:1; a p ro x im a d a m e n te 40:1, a p ro x im a d a m e n te 30:1, a p ro x im a d a m e n te 20 :1 , a p ro x im a d a m e n te 10:1, a p ro x im a d a m e n te 1:1, a p ro x im a d a m e n te 1:2; a p ro x im a d a m e n te 1:3, a p ro x im a d a m e n te 1:5 o a p ro x im a d a m e n te 1:10.

In one respect, the co r p e s ic io de de riva dos de ac se s g ra sos com p ren of a m ezc that of de riva two which includes ac se c a se g ra es lib ra. In one respect, the power in terms of accrued security is at least 50%, for example, at least in the year. im adamen te 55%, ap ro x im adamen te 60%, ap ro x im adamen te 65%, ap ro x im adamen te 70%, ap ro x im adamen te 75%, ap ro x im adamen te 80% , ap ro x im adamen te 85% or ap ro x im adamen te 90%. In an asp e te rm in ado, the po rce ta je in weight of this te rg ra so p ro du c gone is at least ap ro x im adamen te 0, 5%, po re jem p at least 1%, at least 2%, at least 2%, at least 3%, at least 4%, at least five times 5 %, ap ro x im adamen te 6%, ap ro x im adamen te 7%, ap ro x im adamen te 8%, ap ro x im adamen te 9%, ap ro x im adamen te 10%, ap ro x im adamen te 15%, ap ro x im adamen te 20%, ap ro x im adamen te 25%, ap ro x im adamen te 30%, ap ro x im adamen te 35%, ap ro x im adamen te 40% , ap ro x im adamen te 45% or ap ro x im adamen te 50%. In an asp e ct ad ic iona l, the weight re rtion of de riva dos de acido sg ra sos p ro du c gone sd is tin to s of acidic acids with respect to acids g frees is less than ap ro x im adamen te 60: 1, po re jem p lo, less than ap ro x im adamen te 50: 1; ap ro x im adamen te 40: 1, ap ro x im adamen te 30: 1, ap ro x im adamen te 20: 1, ap ro x im adamen te 10: 1, ap ro x im adamen te 1: 1, ap ro x im adamen te 1: 2; a p ro x im a d a m e n te 1: 3, a p ro x im a d a m e n te 1: 5 o a p ro x im a d a m e n te 1:10.

En un asp ec to , la co m p o s ic io n de d e riva d o s de ac ido s g ra sos inc luye uno o m as es te re s g ra sos s e le cc io n a d o s de: d e ca n o a to de etilo , d o d e ca n o a to de e tilo , tr id e c a n o a to de etilo , te tra d e c a n o a to de etilo , p e n ta d e ca n o a to de e tilo , cis-9 -h e xa d e ce n o a to de etilo , h e xa d e ca n o a to de etilo , h e p ta d e ca n o a to de e tilo , c is -11 -o c ta d e ce n o a to de etilo, o c ta d e ca n o a to de etilo, d e ca n o a to de m etilo , do d e ca n o a to de m etilo , tr id e ca n o a to de m etilo , te tra d e c a n o a to de m etilo , pe n ta d e ca n o a to de m etilo , cis-9 -h e x a d e c e n o a to de m etilo , h e xa d e ca n o a to de m etilo , he p ta d e ca n o a to de m etilo , c is -11 -o c ta d e ce n o a to de m etilo , o c ta d e ca n o a to de m etilo , o una co m b in a c io n de los m ism os.

In an asp e ct, the de riva two of ac a se g ra es inc ra es one of these re sg ra are said to be: from an ethyl, from a ca n To the tilo, tr id ecanoa to ethyl, te decanoa to ethyl, pen de ca noa to de e tilo, cis- 9 -he xa de ce noa to ethyl, he xa to ca noa to ethyl , hepta of ethyl tea, c is -11-o of ethyl kettle, octal of ethyl alcohol, of ethyl kettle, of ethyl kettle , because it is methyl, it has been used as ethyl, such as ethyl, cis -9-hexadecene of ethyl, as with methyl, He has a category of ethyl, that is 11% of a ethyl group, or a category of ethyl, or a combination of the same.

En un aspecto, la composicion de derivados de acidos grasos incluye uno o mas acidos grasos libres seleccionados de: acido octanoico, acido decanoico, acido dodecanoico, acido tetradecanoico, acido pentadecanoico, acido cis-9-hexadecenoico, acido hexadecanoico, acido cis-11-octadecenoico, o combinaciones de los mismos.

In one aspect, the fatty acid derivative composition includes one or more free fatty acids selected from: octanoic acid, decanoic acid, dodecanoic acid, tetradecanoic acid, pentadecanoic acid, cis-9- hexadecenoic acid, hexadecanoic acid, cis-11 -octadecenoic, or combinations thereof.

Las composiciones que comprenden los derivados de acidos grasos de la divulgacion pueden utilizarse como combustibles. Por ejemplo, los derivados de acidos grasos pueden utilizarse como, o como un componente de, un biodiesel, un alcohol graso, un ester graso, un triacilglicerido, una gasolina, un diesel, o un combustible de avion. En un motor de combustion interna puede utilizarse una composicion de gasolina o de biodiesel. En un motor a reaccion puede utilizarse un combustible de aviacion. Por consiguiente, en el presente documento se desvelan composiciones de combustible que comprenden los derivados de acidos grasos preparados segun las presentes divulgaciones.

Compositions comprising the fatty acid derivatives of the disclosure can be used as fuels. For example, fatty acid derivatives can be used as, or as a component of, a biodiesel, a fatty alcohol, a fatty ester, a triacylglyceride, a gasoline, a diesel, or an airplane fuel. A gasoline or biodiesel composition can be used in an internal combustion engine. An aviation fuel can be used in a jet engine. Accordingly, fuel compositions are disclosed herein comprising the fatty acid derivatives prepared according to the present disclosures.

Las composiciones que comprenden derivados de acidos grasos de la divulgacion pueden utilizarse como aditivos para combustibles. Por ejemplo, se pueden anadir a un diesel o biodiesel basado en petroleo para mejorar su contenido de combustible renovable, lubricidad, viscosidad cinematica, numero de acido, punto de ebullicion, estabilidad oxidativa, punto de obstruccion del filtro en frio, perfiles de impurezas, nivel de ceniza sulfatada, numero de cetano, punto de enturbamiento o punto de fluidez. Por consiguiente, tambien se desvelan composiciones de aditivos para combustible que comprenden derivados de acidos grasos producidos segun las presentes divulgaciones.

Compositions comprising fatty acid derivatives of the disclosure can be used as fuel additives. For example, they can be added to a petroleum-based diesel or biodiesel to improve their renewable fuel content, lubricity, kinematic viscosity, acid number, boiling point, oxidative stability, cold filter blockage point, impurity profiles, sulfated ash level, cetane number, cloud point or pour point. Accordingly, compositions of fuel additives comprising fatty acid derivatives produced according to the present disclosures are also disclosed.

Las composiciones que comprenden derivados de acidos grasos de la divulgacion tambien pueden utilizarse como composiciones de biocrudo, que pueden servir como materia prima para producir otros compuestos derivados del petroleo. Por ejemplo, hidrocarburos de cadena larga, olefinas internas o terminales, alcanos, aldehidos grasos y esteres grasos preparados segun la presente divulgacion pueden procesarse adicionalmente para producir combustibles, aditivos de combustible, aleaciones de combustible y/o productos quimicos. Por consiguiente, se describen composiciones de biocrudo que comprenden derivados de acidos grasos preparados segun las presentes divulgaciones.

Compositions comprising fatty acid derivatives of the disclosure can also be used as biocrude compositions, which can serve as raw material to produce other petroleum-derived compounds. For example, long chain hydrocarbons, internal or terminal olefins, alkanes, fatty aldehydes and fatty esters prepared according to the present disclosure can be further processed to produce fuels, fuel additives, fuel alloys and / or chemicals. Accordingly, biocrude compositions comprising fatty acid derivatives prepared according to the present disclosures are described.

Las composiciones que comprenden derivados de acidos grasos de la divulgacion pueden utilizarse como materias primas en la fabricacion de detergentes y tensioactivos, complementos nutricionales, polimeros, reemplazos de parafina, lubricantes, disolventes, productos para el cuidado personal, aditivos de procesamiento de caucho, inhibidores de corrosion, emulsionantes, plasticos, tejidos, cosmeticos, productos de papel, recubrimientos, liquidos para la metalurgia, dielectricos, engrasantes y/o emolientes. Por consiguiente, tambien se desvelan composiciones de materia prima que comprenden derivados de acidos grasos preparados segun las presentes divulgaciones.

Compositions comprising fatty acid derivatives of the disclosure can be used as raw materials in the manufacture of detergents and surfactants, nutritional supplements, polymers, paraffin replacements, lubricants, solvents, personal care products, rubber processing additives, inhibitors of corrosion, emulsifiers, plastics, fabrics, cosmetics, paper products, coatings, metallurgy liquids, dielectrics, greasers and / or emollients. Accordingly, raw material compositions comprising fatty acid derivatives prepared according to the present disclosures are also disclosed.

Descripcion de las figuras

Description of the figures

La figura 1 (FIG. 1) es una tabla que identifica diversos genes que pueden sobreexpresarse o atenuarse para aumentar la produccion de derivados de acidos grasos. La tabla tambien identifica varios genes que pueden modularse para alterar la estructura del producto derivado de acido graso. Algunos de los genes que se utilizan para alterar la estructura del derivado de acido graso tambien aumentaran la produccion de derivados de acido graso.

Figure 1 (FIG. 1) is a table that identifies various genes that can be overexpressed or attenuated to increase the production of fatty acid derivatives. The table also identifies several genes that can be modulated to alter the structure of the fatty acid derivative product. Some of the genes that are used to alter the structure of the fatty acid derivative will also increase the production of fatty acid derivatives.

La figura 2 (FIG. 2) es un diagrama que ilustra la ruta de la betaoxidacion, incluyendo etapas catalizadas por las siguientes enzimas (1) acil-CoA sintasa (EC 6.2.1.-). (2) acil-CoA deshidrogenasa (EC 1.3.99.3), (3) enoil-CoA hidratasa (EC 4.2.1.17); (4) 3-hidroxibutiril-CoA epimerasa (EC 5.1.2.3) y (5) 3-cetoacil-CoA tiolasa (e C 2.3.1.16). Esta reaccion final del ciclo de la p-oxidacion, libera acetil-CoA y un acido graso acil-CoA dos carbonos mas corto, listos para pasar nuevamente por las reacciones de la p-oxidacion.

Figure 2 (FIG. 2) is a diagram illustrating the route of beta oxidation, including steps catalyzed by the following enzymes (1) acyl-CoA synthase (EC 6.2.1.-). (2) acyl-CoA dehydrogenase (EC 1.3.99.3), (3) enoyl-CoA hydratase (EC 4.2.1.17); (4) 3-hydroxybutyryl-CoA epimerase (EC 5.1.2.3) and (5) 3-ketoacyl-CoA thiolase (e C 2.3.1.16). This final reaction of the p-oxidation cycle releases acetyl-CoA and a shorter two-carbon acyl-CoA fatty acid, ready to go through the p-oxidation reactions again.

La figura 3 (FIG. 3) es un diagrama que ilustra la ruta biosintetica de la FAS (Fatty Acid Synthase, acido graso sintasa).

Figure 3 (FIG. 3) is a diagram illustrating the biosynthetic pathway of FAS ( Fatty Acid Synthase, fatty acid synthase).

La figura 4 (FIG. 4) es un diagrama que ilustra las rutas biosinteticas que producen esteres grasos dependiendo de los sustratos proporcionados.

Figure 4 (FIG. 4) is a diagram illustrating the biosynthetic pathways that produce fatty esters depending on the substrates provided.

La figura 5 (FIG. 5) es un diagrama que ilustra las rutas biosinteticas que producen alcoholes grasos.

Figure 5 (FIG. 5) is a diagram illustrating the biosynthetic pathways that produce fatty alcohols.

La figura 6 (FIG. 6) es un grafico que representa la produccion de alcohol graso por la cepa transformada conjuntamente con pCDFDuet-1-fadD-acr1 y plasmidos que contienen diversos genes de tioesterasa. Se identificaron acidos grasos C10, C12, C14, C16 y C-^saturados.

Figure 6 (FIG. 6) is a graph depicting the production of fatty alcohol by the strain transformed together with pCDFDuet-1-fadD-acr1 and plasmids containing various thioesterase genes. C10, C12, C14, C16 and C- ^ saturated fatty acids were identified.

La figura 7 (FIG. 7) es un grafico que representa la produccion de alcohol graso mediante la cepa descrita en el Ejemplo 3, transformado conjuntamente con pCDFDuet-1 -fadD-acrl y plasmidos que contienen diversos genes de tioesterasa. Las cepas se cultivaron en condiciones aerobias a 25 °C o a 37 °C en un medio mineral M9 que contenia glucosa al 0,4 % en matraces de agitacion. Los alcoholes grasos se detectaron en los sedimentos celulares, asi como en los sobrenadantes, lo que indica una produccion extracelular sustancial de dichos alcoholes. El cultivo a 25 °C produjo la liberacion de aproximadamente 25 % del producto de las celulas, mientras que el cultivo a 37 °C produjo la liberacion de aproximadamente 50 % del producto de la celula.

Figure 7 (FIG. 7) is a graph depicting the production of fatty alcohol by the strain described in Example 3, transformed together with pCDFDuet-1 -fadD-acrl and plasmids containing various thioesterase genes. The strains were grown under aerobic conditions at 25 ° C or 37 ° C in an M9 mineral medium containing 0.4% glucose in shake flasks. Fatty alcohols were detected in cell sediments, as well as in supernatants, indicating a substantial extracellular production of said alcohols. The culture at 25 ° C produced the release of approximately 25% of the product from the cells, while the culture at 37 ° C produced the release of approximately 50% of the product from the cell.

Las figuras 8A-D (FIGs. 8A-D) son graficos que representan espectros de GC-MS (Cromatografia de gases-Espectrometria de masas) de octanoato de octilo (C8C8) producido por un hospedador de produccion que expresa alcohol acetil transferasa (AAT, EC 2.3.1.84) y hospedadores de produccion que expresan ester sintasa (EC 2.3.1.20, 2.3.1.75). La FIG. 8A es un espectro de GC-MS que muestra un extracto de acetato de etilo de la cepa C41 (DE3,Afad/pHZ1.43)/pRSET B+pAS004.114B) en donde el plasmido pHZ1.43 expresaba ADP1 ester sintasa (EC 2.3.1.20, 2.3.1.75). La FIG. 8 B es un espectro de GC-MS que muestra un extracto de acetato de etilo de la cepa C41 (DE3, AfadE/pHZ1.43)/pRSET B+pAS004.114B) en donde el plasmido pHZ1.43 expresaba SAAT. La FIG. 8 C es un espectro de GC-MS que muestra un extracto de acetato de acetilo de la cepa C41 (DE3, AfadE/pHZ1.43)/pRSET B+pAS004.114B) en donde el plasmido pHZ1.43 no contenia ADP1 (una ester sintasa) o SAAT. La FIG. 8 D es un espectro de GC-MS que muestra el espectro de masas y el patron de fragmentacion de C8C8 producido por la cepa C41 (DE3, AfadE1/pHZ1.43)/pRSET B+pAS004.114B en donde el plasmido pHZ1.43 expreso SAAT).

Figures 8A-D (FIGs. 8A-D) are graphs depicting GC-MS (Gas Chromatography-Mass Spectrometry) spectra of octyl octanoate (C8C8) produced by a production host expressing alcohol acetyl transferase (AAT , EC 2.3.1.84) and production hosts expressing ester synthase (EC 2.3.1.20, 2.3.1.75). FIG. 8A is a spectrum of GC-MS showing an ethyl acetate extract of strain C41 (DE3, Afad / pHZ1.43) / pRSET B + pAS004.114B) where the plasmid pHZ1.43 expressed ADP1 ester synthase (EC 2.3.1.20, 2.3.1.75). FIG. 8 B is a spectrum of GC-MS showing an ethyl acetate extract of strain C41 (DE3, AfadE / pHZ1.43) / pRSET B + pAS004.114B) wherein the plasmid pHZ1.43 expressed SAAT. FIG. 8 C is a spectrum of GC-MS showing an extract of acetyl acetate of strain C41 (DE3, AfadE / pHZ1.43) / pRSET B + pAS004.114B) where plasmid pHZ1.43 did not contain ADP1 (a ester synthase) or SAAT. FIG. 8 D is a GC-MS spectrum that shows the mass spectrum and fragmentation pattern of C8C8 produced by strain C41 (DE3, AfadE1 / pHZ1.43) / pRSET B + pAS004.114B where the plasmid pHZ1.43 SAAT expressed).

La Figura 9 (FIG. 9) es un grafico que representa la distribucion de esteres etilicos elaborados (segun el Ejemplo 9) cuando la ester sintasa de A. baylyi ADP1 (WSadpl) se expreso conjuntamente con una tioesterasa de Cuphea hookeriana en un hospedador de produccion.

Figure 9 (FIG. 9) is a graph depicting the distribution of elaborated ethyl esters (according to Example 9) when the A. baylyi ADP1 synthase ester (WSadpl) was expressed in conjunction with a Cuphea hookeriana thioesterase in a host of production.

La Figura 10 (FIG. 10) es un grafico que representa la produccion de esteres etilicos por diversas ester sintasas a 25 °C. Los esteres etilicos se produjeron mediante cepas de E. coli recombinantes portadoras de diversos genes de ester sintasa. Las cepas recombinantes fueron (1) C41 (DE3, AfadEAfabR)/pETDuet-1-'TesA+pCDFDuet-1-fadD con 1 pHZ1.43; (2) pHZ1.97_377; (3) pHZ1.97_atfA2; (4) pHZ1.97_376; (5) pHZ1.97_atfA1; y (6) sin plasmidos (control).

Figure 10 (FIG. 10) is a graph depicting the production of ethyl esters by various ester synthases at 25 ° C. The ethyl esters were produced by recombinant E. coli strains carrying various ester synthase genes. The recombinant strains were (1) C41 (DE3, AfadEAfabR) / pETDuet-1-'TesA + pCDFDuet-1-fadD with 1 pHZ1.43; (2) pHZ1.97_377; (3) pHZ1.97_atfA2; (4) pHZ1.97_376; (5) pHZ1.97_atfA1; and (6) without plasmids (control).

La Figura 11 (FIG. 11) es un grafico que representa la composicion de acilo de los esteres etilicos de acidos grasos (FAEE, fatty acid ethyl esters) producidos a partir de diversas cepas de E. coli. Las cepas recombinantes son (1) C41 (DE3, AfadEAfabR)/pETDuet-1-'TesA pCDFDuet-1 -fadD con 1 pHZ1.43; (2) pHZ1.97_377; (3) pHZ1.97_atfA2; (4) pHZ1.97_376; (5) pHZ1.97_atfA1; y (6) sin plasmidos (control).

Figure 11 (FIG. 11) is a graph depicting the acyl composition of fatty acid ethyl esters (FAEE ) produced from various strains of E. coli. Recombinant strains are (1) C41 (DE3, AfadEAfabR) / pETDuet-1-'TesA pCDFDuet-1 -fadD with 1 pHZ1.43; (2) pHZ1.97_377; (3) pHZ1.97_atfA2; (4) pHZ1.97_376; (5) pHZ1.97_atfA1; and (6) without plasmids (control).

La Figura 12 (FIG. 12) es un grafico que representa la produccion de esteres etilicos mediante diversas ester sintasas a 37 °C. Los esteres etilicos se produjeron mediante cepas de E. coli recombinantes portadoras de diversos genes de ester sintasa. Las cepas recombinantes fueron (1) C41 (DE3, AfadEAfabR)/pETDuet-1-'TesA + pCDFDuet-1-fadD con 1 pHZ1.43; (2) pHZ1.97_377; (3) pHZ1.97_atfA2; (4) pHZ1.97_376; (5) pHZ1.97_atfA1; y (6) sin plasmidos (control).

Figure 12 (FIG. 12) is a graph depicting the production of ethyl esters by various ester synthases at 37 ° C. The ethyl esters were produced by recombinant E. coli strains carrying various ester synthase genes. The recombinant strains were (1) C41 (DE3, AfadEAfabR) / pETDuet-1-'TesA + pCDFDuet-1-fadD with 1 pHZ1.43; (2) pHZ1.97_377; (3) pHZ1.97_atfA2; (4) pHZ1.97_376; (5) pHZ1.97_atfA1; and (6) without plasmids (control).

La Figura 13 (FIG. 13) es un grafico que muestra las concentraciones de acidos grasos libres (FFA, free fatty acids) y esteres etilicos de acidos grasos (FAEE, fatty acid ethyl esters) producidos a partir de tres colonias individuales de los transformantes, C41 (DE3, AfadEAfabR)/ pETDuet-1-'TesA+pCD-FDuet-1-fadD pHZ1.97_atfA2. El FFA se transformo en ester etilico de acido graso (FAEE) y se cuantifico mediante GC/MS. La Figura 14 (FIG. 14) es un diagrama que representa las regiones de control de FabA (SEC ID NO: 33) y FabB (SEC ID NO: 34). Los sitios de union consenso de FadR y FabR se muestran en negrita. Las flechas verticales indican las posiciones donde pueden realizarse mutaciones para alterar la expresion de fabA. La base propuesta para cada posicion tambien se indica entre parentesis. Las dos regiones que constituyen las regiones -35 y -10 del promotor tipico de E. coli, se indican entre parentesis. Tambien se muestran las mutaciones propuestas que acercan al promotor a la secuencia promotora consenso.

Figure 13 (FIG. 13) is a graph showing the concentrations of free fatty acids (FFA ) and fatty acid ethyl esters (FAEE ) produced from three individual colonies of the transformants , C41 (DE3, AfadEAfabR) / pETDuet-1-'TesA + pCD-FDuet-1-fadD pHZ1.97_atfA2. FFA was transformed into fatty acid ethyl ester (FAEE) and quantified by GC / MS. Figure 14 (FIG. 14) is a diagram representing the control regions of FabA (SEQ ID NO: 33) and FabB (SEQ ID NO: 34). The consensus binding sites of FadR and FabR are shown in bold. The vertical arrows indicate the positions where mutations can be made to alter the expression of fabA. The proposed basis for each position is also indicated in parentheses. The two regions that constitute the -35 and -10 regions of the typical E. coli promoter are indicated in parentheses. The proposed mutations that bring the promoter to the consensus promoter sequence are also shown.

Las figuras 15A-B (FIGs. 15A-B) son cromatogramas que representan analisis de GC/MS. La FIG. 15A es un cromatograma que representa los componentes de un extracto de acetato de etilo del cultivo de la cepa LS9001 de E. coli transformada con plasmidos pCD-FDuet-1-fadD-WSadp1, pETDuet-1-'TesA. La FIG. 15B es un cromatograma que representa el hexadecanoato de etilo y el oleato de etilo, que se utilizaron como referencias. La Figura 16 (FIG. 16) es un mapa del plasmido pOP-80.

Figures 15A-B (FIGs. 15A-B) are chromatograms representing GC / MS analysis. FIG. 15A is a chromatogram representing the components of an ethyl acetate extract from the culture of E. coli strain LS9001 transformed with plasmids pCD-FDuet-1-fadD-WSadp1, pETDuet-1-'TesA. FIG. 15B is a chromatogram representing ethyl hexadecanoate and ethyl oleate, which were used as references. Figure 16 (FIG. 16) is a map of plasmid pOP-80.

La Figura 17 (FIG. 17) es la secuencia de ADN completa del plasmido pOP-80 (SEC ID NO: 1)

Figure 17 (FIG. 17) is the complete DNA sequence of plasmid pOP-80 (SEQ ID NO: 1)

La Figura 18 (FIG. 18 ) es la secuencia de ADN (s Ec ID n O: 2) del gen fadD35 (n° de registro NP_217021 de GenBank) de E. coli optimizada con codones.

Figure 18 (FIG. 18) is the DNA sequence (s Ec ID n O: 2) of the fadD35 gene (GenBank registration number NP_217021) of E. coli optimized with codons.

La Figura 19 (FIG. 19) es la secuencia de ADN (SEC ID NO: 3) del gen fadDI (n° de registro NP_251989 de GenBank) de E. coli optimizada con codones.

Figure 19 (FIG. 19) is the DNA sequence (SEQ ID NO: 3) of the fadDI gene (GenBank registration number NP_251989) of E. coli optimized with codons.

La Figura 20 (FIG. 20) es el cebador BsyhfLBspHIF (SEQ ID NO: 4) basado en la secuencia de ADN depositada en el NCBI con el n° de registro NC_000964 de GenBank.

Figure 20 (FIG. 20) is the BsyhfLBspHIF primer (SEQ ID NO: 4) based on the DNA sequence deposited in the NCBI with GenBank registration number NC_000964.

La Figura 21 (FIG. 21) es el cebador BsyhfLEcoR (SEQ ID NO: 5) basado en la secuencia de ADN depositada en el NCBI con el n° de registro NC_000964 de GenBank.

Figure 21 (FIG. 21) is the BsyhfLEcoR primer (SEQ ID NO: 5) based on the DNA sequence deposited in the NCBI with GenBank registration number NC_000964.

La Figura 22 (FIG. 22) es la secuencia de ADN (SEQ ID NO: 6) del gen yhfL de Bacillus subtilis.

Figure 22 (FIG. 22) is the DNA sequence (SEQ ID NO: 6) of the yhfL gene of Bacillus subtilis.

La Figura 23 (FIG. 23) es el cebador Scfaa3pPciF (SEQ ID NO: 7) basado en la secuencia de ADN depositada en el NCBI con el n° de registro NC_001141 de GenBank.

Figure 23 (FIG. 23) is the Scfaa3pPciF primer (SEQ ID NO: 7) based on the DNA sequence deposited in the NCBI with GenBank registration number NC_001141.

La Figura 24 (FIG. 24) es el cebador Scfaa3pPciI (SEQ ID NO: 8) basado en la secuencia de ADN depositada en el NCBI con el n° de registro NC_001141 de GenBank.

Figure 24 (FIG. 24) is the Scfaa3pPciI primer (SEQ ID NO: 8) based on the DNA sequence deposited in the NCBI with GenBank registration number NC_001141.

La Figura 25 (FIG. 25) es la secuencia de ADN (SEQ ID NO: 9) del gen faa3 de Saccharomyces cerevisiae (n° de registro NP_012257 de GenBank).

Figure 25 (FIG. 25) is the DNA sequence (SEQ ID NO: 9) of the faa3 gene of Saccharomyces cerevisiae (GenBank registration number NP_012257).

La Figura 26 (FIG. 26) es el cebador Smprk59BspF (SEQ ID NO: 10) basado en la secuencia de ADN depositada en el NCBI con el n° de registro n Z_AAVz 01000044 de GenBank.

Figure 26 (FIG. 26) is the primer Smprk59BspF (SEQ ID NO: 10) based on the DNA sequence deposited in the NCBI with the registration number n Z_AAVz 01000044 of GenBank.

La Figura 27 (FIG. 27) es el cebador Smprk59HindR (SEQ ID NO: 11) basado en la secuencia de ADN depositada en el NCBI con el n° de registro n Z_AAVZ01000044 de GenBank.

Figure 27 (FIG. 27) is the Smprk59HindR primer (SEQ ID NO: 11) based on the DNA sequence deposited in the NCBI with the registration number n Z_AAVZ01000044 of GenBank.

La Figura 28 (FIG. 28) es el cebador PrkBsp (SEC ID NO: 12).

Figure 28 (FIG. 28) is the PrkBsp primer (SEQ ID NO: 12).

La Figura 29 (FIG. 29) es la secuencia de ADN que codifica la proteina ZP_01644857 de Stenotrophomonas maltophilia R551-3 (SEC ID NO: 13).

Figure 29 (FIG. 29) is the DNA sequence encoding Stenotrophomonas maltophilia R551-3 protein ZP_01644857 (SEQ ID NO: 13).

La Figura 30 (FIG. 30) es la secuencia de proteinas de ZP_01644857 de Stenotrophomonas maltophilia ATCC 17679 (SEC ID N°: 14).

Figure 30 (FIG. 30) is the protein sequence of ZP_01644857 from Stenotrophomonas maltophilia ATCC 17679 (SEQ ID NO: 14).

La figura 31 (FIG. 31) Es un esquema de una nueva ruta para la produccion de aldehido graso.

Figure 31 (FIG. 31) is a schematic of a new route for the production of fatty aldehyde.

La figura 32 (FIG. 32) es un listado de la secuencia de nucleotidos (SEC ID NO: 15) y de la secuencia de aminoacidos correspondiente (SEC ID NO: 16) del gen carde Nocardia sp. NRRL 5646.

Figure 32 (FIG. 32) is a listing of the nucleotide sequence (SEQ ID NO: 15) and the sequence of Corresponding amino acids (SEQ ID NO: 16) of the cardinal Nocardia sp. NRRL 5646.

La figura 33 (FIG. 33) es un listado de motivos de secuencia de aminoacidos de homologos de CAR.

Figure 33 (FIG. 33) is a list of amino acid sequence motifs of CAR homologs.

Las figuras 34A-B (FIGs. 34A-B) son trazados de GC/MS de olefinas producidas por celulas de Jeotgalicoccus sp.ATCC 8456 y celulas de Jeotgalicoccus halotolerans DSMZ 17274, respectivamente.

Figures 34A-B (FIGs. 34A-B) are GC / MS traces of olefins produced by Jeotgalicoccus sp. ATCC 8456 cells and Jeotgalicoccus halotolerans DSMZ 17274 cells, respectively.

Las figuras 35A-B (FIGs. 35A-B) son trazados de GC/MS de olefinas producidas por celulas de Jeotgalicoccus pinnipedalis DSMZ 17030 y celulas de Jeotgalicoccus psychrophilus DSMZ 19085, respectivamente.

Figures 35A-B (FIGs. 35A-B) are GC / MS traces of olefins produced by Jeotgalicoccus pinnipedalis DSMZ 17030 cells and Jeotgalicoccus psychrophilus DSMZ 19085 cells, respectively.

Las figuras 36A-B (FIGs. 36A-B) son patrones de fragmentacion de espectrometria de masas de dos a-olefinas producidas por celulas de Jeotgalicoccus ATCC 8456. El compuesto A se identifico como 1-nonadeceno y el compuesto B como 18-metil-1-nonadeceno.

Figures 36A-B (FIGs. 36A-B) are mass spectrometry fragmentation patterns of two a-olefins produced by Jeotgalicoccus ATCC 8456 cells. Compound A was identified as 1-nonadecene and compound B as 18-methyl -1-nonadecene.

La figura 37 (FIG. 37) es un esquema de un analisis filogenetico de ARNr 16s de Jeotgalicoccus ATCC 8456. Las figuras 38A-B (FIGs. 38A-B) son trazados de GC/MS de a-olefinas producidas por celulas de Jeotgalicoccus sp. ATCC 8456 despues de suministrar acido eicosanoico (FIG. 38A) o acido estearico (FIG.

Figure 37 (FIG. 37) is a schematic of a phylogenetic analysis of 16s rRNA from Jeotgalicoccus ATCC 8456. Figures 38A-B (FIGs. 38A-B) are GC / MS traces of a-olefins produced by Jeotgalicoccus cells sp. ATCC 8456 after supplying eicosanoic acid (FIG. 38A) or stearic acid (FIG.

La Figura 39 (FIG. 39) es un trazado de GC/MS de a-olefinas (1-heptadeceno) producidas por lisados acelulares de celulas de Jeotgalicoccus sp. ATCC 8456, en comparacion con un trazado de lisado acelular sin el sustrato de acidos grasos C18 y un trazado del propio sustrato de acidos grasos C18.

Figure 39 (FIG. 39) is a GC / MS plot of a-olefins (1-heptadecene) produced by acellular lysates of Jeotgalicoccus sp. ATCC 8456, compared to an acellular lysate path without the C18 fatty acid substrate and a C18 fatty acid substrate path itself.

La Figura 40 (FIG. 40) es una representacion digital de un gel de SDS-PAGE de una fraccion proteica de produccion final de a-olefinas purificadas de celulas de Jeotgalicoccus sp. ATCC 8456.

Figure 40 (FIG. 40) is a digital representation of an SDS-PAGE gel of a final production protein fraction of purified a-olefins from Jeotgalicoccus sp. ATCC 8456.

Las figuras 41A-B (FIGs. 41A-B) son secuencias de nucleotidos (SEC ID NO: 25) y de aminoacidos (SEC ID NO: 26) de la orf880, respectivamente. La FIG. 41C es la secuencia parcial de ARNr 16s (SEQ ID NO: 27) de Jeotgalicoccus sp. ATCC8456.

Figures 41A-B (FIGs. 41A-B) are nucleotide (SEQ ID NO: 25) and amino acid (SEQ ID NO: 26) sequences of orf880, respectively. FIG. 41C is the partial sequence of 16s rRNA (SEQ ID NO: 27) of Jeotgalicoccus sp. ATCC8456.

La Figura 42 (FIG. 42) es un trazado de GC/MS de a-olefinas producidas por E. coli despues de la expresion de 8456_orf880 de Jeotgalicoccus sp. y el suministro de acido estearico.

Figure 42 (FIG. 42) is a GC / MS plot of a-olefins produced by E. coli after 8456_orf880 expression of Jeotgalicoccus sp. and the supply of stearic acid.

La Figura 43 (FIG. 43) es un esquema de un analisis de remuestreo (bootstrap) filogenetico de homologos de 8456_orf880 utilizando el programa ClustalW.

Figure 43 (FIG. 43) is a schematic of a phylogenetic restraint analysis ( bootstrap) of homologs of 8456_orf880 using the ClustalW program.

La figura 44 (FIG. 44) describe motivos de aminoacidos para identificar tioesterasas precursoras utiles en la presente divulgacion.

Figure 44 (FIG. 44) describes amino acid motifs to identify precursor thiosterases useful in the present disclosure.

Las figuras 45A-B (FIGs. 45A-B) incluyen tablas que enumeran los resultados de ensayos que identifican tioesterasas mutantes con propiedades alteradas. En particular, la FIG. 45A incluye listados de mutantes con puntuaciones Z con un valor de al menos 3 para la actividad (es decir, velocidad catalitica) con respecto al sustrato mencionado o especificidad para el sustrato mencionado; y la FIG. 45B es una tabla de mutantes que tienen un rendimiento/produccion mejorado y/o aumentado de derivados de acidos grasos con puntuaciones Z con un valor de al menos 3.

Figures 45A-B (FIGs. 45A-B) include tables that list the results of assays that identify mutant thiosterases with altered properties. In particular, FIG. 45A includes listings of mutants with Z scores with a value of at least 3 for the activity (ie, catalytic velocity) with respect to the mentioned substrate or specificity for the mentioned substrate; and FIG. 45B is a table of mutants that have an improved and / or increased yield / production of fatty acid derivatives with Z scores with a value of at least 3.

Las figuras 46A-E (FIGs. 46A-E) incluyen tablas que enumeran los resultados de ensayos que identifican tioesterasas mutantes con rendimiento proporcional alterado de esteres grasos frente a otros productos (por ejemplo, derivados de acidos grasos distintos de esteres grasos). En particular, la FIG. 46A es una tabla que muestra mutantes que tienen puntuaciones Z con un valor de al menos 3 con respecto al rendimiento proporcional o porcentual de esteres grasos frente a acidos grasos libres. La FIG. 46B es una tabla que muestra mutantes que tienen puntuaciones Z con un valor inferior a -3 con respecto al rendimiento proporcional o porcentual de esteres grasos frente a acidos grasos libres. La FIG. 46C es una tabla que muestra mutantes que tienen puntuaciones Z con un valor de al menos 3 con respecto al rendimiento in vivo de derivados de acidos grasos. La FIG. 46D es una tabla que muestra mutantes que tienen puntuaciones Z con un valor de al menos 3 con respecto al rendimiento proporcional de derivados de acidos grasos de cadena corta (por ejemplo, C8, Cg, C10, C11, C12, C13 y/o C14) frente a otros derivados de acidos grasos (por ejemplo, derivados de acidos grasos distintos de derivados de acidos grasos de cadena corta incluyendo, por ejemplo, derivados de acidos grasos de cadena larga (por ejemplo, C15, C16, C17, C18, C1gy/o C20)). La FIG. 46E es una tabla que muestra mutantes que tienen puntuaciones Z con un valor inferior a -3 con respecto al rendimiento proporcional de derivados de acidos grasos de cadena corta (por ejemplo, C8, Cg, C10, C11, C12, C13 y/o C14) frente a otros derivados de acidos grasos (por ejemplo, derivados de acidos grasos distintos de derivados de acidos grasos de cadena corta incluyendo, por ejemplo, derivados de acidos grasos de cadena larga (por ejemplo, C15, C16, C17, C18, C1gy/o C20)).

Figures 46A-E (FIGs. 46A-E) include tables that list the results of assays that identify mutant thiosterases with proportionally altered yield of fatty esters against other products (eg, fatty acid derivatives other than fatty esters). In particular, FIG. 46A is a table showing mutants that have Z scores with a value of at least 3 with respect to the proportional or percentage yield of fatty esters versus free fatty acids. FIG. 46B is a table showing mutants that have Z scores with a value below -3 with respect to the proportional or percentage yield of fatty esters versus free fatty acids. FIG. 46C is a table showing mutants that have Z scores with a value of at least 3 with respect to the in vivo performance of fatty acid derivatives. FIG. 46D is a table showing mutants that have Z scores with a value of at least 3 with respect to the proportional yield of short-chain fatty acid derivatives (for example, C8, Cg, C10, C11, C12, C13 and / or C14 ) versus other fatty acid derivatives (for example, fatty acid derivatives other than short chain fatty acid derivatives including, for example, long chain fatty acid derivatives (eg, C15, C16, C17, C18, C1gy / or C20)). FIG. 46E is a table showing mutants that have Z scores with a value less than -3 with respect to the proportional yield of short-chain fatty acid derivatives (for example, C8, Cg, C10, C11, C12, C13 and / or C14 ) versus other fatty acid derivatives (for example, fatty acid derivatives other than short chain fatty acid derivatives including, for example, long chain fatty acid derivatives (eg, C15, C16, C17, C18, C1gy / or C20)).

La Figura 47 (FIG. 47) es un alineamiento de secuencias de homologos de 'TesA utilizando, como secuencia de referencia con fines de numeracion, los restos de aminoacidos de una 'TesA de E. coli (es decir, una TesA sin el peptido senal).

Figure 47 (FIG. 47) is an alignment of 'TesA homolog sequences using, as a reference sequence for numbering purposes, the amino acid residues of an E. coli ' TesA (i.e., a TesA without the peptide signal).

La Figura 48 (FIG. 48) es un grafico que representa los titulos y la composicion de FAME para la cepa MG1655 (AfadE) pTrc-'TesA_fadD.

Figure 48 (FIG. 48) is a graph depicting the titles and composition of FAME for strain MG1655 ( AfadE) pTrc-'TesA_fadD.

La Figura 49 (FIG. 49) es un grafico que representa los titulos y la composicion de FAME para las cepas MG1655 (AfadE) y C41(AfadE) que expresan fadD y 'tesAen plasmidos durante un ciclo de fermentacion de 25 horas.

Figure 49 (FIG. 49) is a graph depicting the titles and composition of FAME for strains MG1655 ( AfadE) and C41 (AfadE) expressing fadD and 'tesAen plasmids during a 25-hour fermentation cycle.

La Figura 50 (FIG. 50) es un grafico que representa los titulos y la composicion de FAME para la cepa MG1655 (AfadE) pTrc-'TesA_fadD.

Figure 50 (FIG. 50) is a graph depicting the titles and composition of FAME for strain MG1655 ( AfadE) pTrc-'TesA_fadD.

La Figura 51 (FIG. 51) es un grafico que representa los titulos y la composicion de FAME para las cepas MG1655 (AfadE) y C41 (AfadE) que expresan fadD y 'tesA en plasmidos durante un ciclo de fermentacion de 25 horas.

Figure 51 (FIG. 51) is a graph depicting the titles and composition of FAME for strains MG1655 ( AfadE) and C41 ( AfadE) expressing fadD and 'tesA in plasmids during a 25-hour fermentation cycle.

La Figura 52 (FIG. 52) es un grafico que representa los titulos y la composicion de FFA para las cepas MG1655 (AfadE) y C41 (AfadE) que expresan fadD y 'tesA en plasmidos durante un ciclo de fermentacion de 25 horas. La Figura 53 (FIG. 53) es un grafico que muestra los titulos de FAME para las cepas MG1655 (AfadE) que exp resa n 'tesA de E. coli 'tesA de P. luminescens, 'tesA de V. harveyi y tesB de P. profundum en p lasm idos, d u ra n te un c ic lo de fe rm e n ta c io n de 24 horas. Los titu lo s se rep re se n ta n en m g/l y m g/l/D O .

Figure 52 (FIG. 52) is a graph depicting the titles and composition of FFA for strains MG1655 ( AfadE) and C41 ( AfadE) expressing fadD and 'tesA in plasmids during a 25-hour fermentation cycle. Figure 53 (FIG. 53) is a graph showing the FAME titles for MG1655 ( AfadE) strains that Exp. n 'tesA of E. coli' tesA of P. luminescens, 'tesA of V. harveyi and tesB of P. profundum in plasmids, during a period of faith in the ta 24 hours. The titles are represented in mg / lymg / l / OD.

La Figura 54 (FIG. 54) es un g ra fico de titu lo s de F F A pa ra las ce p a s M G 1655 (AfadE) que e xp re sa n 'te sA de E. coli, 'tesA de P. luminescens, 'te sA de V. harveyi y te sB de P. profundum en p lasm idos , d u ran te un c ic lo de fe rm e n ta c io n de 24 horas. Los titu lo s se rep re se n ta n en m g/l (b a rra s ) y m g /l/D O (trian gu los).

Figure 54 (FIG. 54) is a graph of the FFA title for MG 1655 ( AfadE) that is E. coli, p. Of P. luminescens, You know about V. harveyi and you know about P. profundum in plasmids, you will have a faith in rm within 24 hours. The titles are represented in mg / l (ba rra s) and mg / l / OD (triangles).

La Figura 55 (FIG. 55) com p ara las reg io nes de se cu e n c ia re leva n tes de las t io e s te ra sa s na tu ra les que com p re n d e n res tos en p o s ic io ne s que co rre spo nde n a m u ta c io n e s en 'T e sA que in tro du cen p ro p ieda de s a lte rad as . El resto re leva n te se resa lta en co lo r oscu ro y se a line a con res tos co rre sp o n d ie n te s en las tio e s te ra s a s na tu ra les .

Figure 55 (FIG. 55) compares the regions of the type of respondents who are responsible for the conditions in which they are in position. That is the reason why there is no need for the introduction of the radius. The rest of the survey is reviewed in the dark and is aligned with the other sprees in the uncles in your country.

La Figura 56 (FIG. 56) es un lis tado de nu m ero s de reg is tro de G e n B a n k de ho m o lo g o s de 'TesA .

Figure 56 (FIG. 56) is a list of reg numbers from G e n B to n k of ho m or lo g o s of 'TesA.

Las Figuras 57A-C (FIG. 57A-C) son g ra fico s que rep rese n tan e sp ec ific ida d de sus tra to (p u n tu a c io n Z ) fre n te a p o s ic io n e s de res tos de a m in o a c id o s c o rre sp o n d ie n te s a la se cu e n c ia de 'T e sA de la S E C ID NO: 31 con s im b o lo s que rep rese n tan n ive les de co n se rva c io n en el a lin e a m ie n to con s70 para e sp e c ific id a d de C 10 (FIG.

Figures 57A-C (FIG. 57A-C) are graphs that represent the extent of their work (punctuated Z) in relation to the rest of the resources. From am in o a c id osco rre sp ond ie n you know the meaning of SEQ ID NO: 31 with only what you know is so important. rva c io n in the lin eam ie n to with s70 for e sp ec ific id ad of C 10 (FIG.

57A), e sp e c ific id a d de C 12 (FIG. 57B) y e sp e c ific id a d de C 14 (FIG. 57C).

57A), e sp e c ific id a d of C 12 (FIG. 57B) and e sp e c ific id a d of C 14 (FIG. 57C).

La Figura 58 (FIG. 58) m ue s tra la s e cu e n c ia de a m in o a c id o s de una 'T e s A de E. coli (S E C ID NO: 31).

Figure 58 (FIG. 58) shows the sequence of am in or of a 'T is A of E. coli (SEQ ID NO: 31).

La Figura 59 (FIG. 59) m ue s tra una se cu e n c ia de nu c le o tid o s que cod ifica una 'T e s A de E. coli (S E C ID NO: 32). La Figura 60 (FIG. 60) es un g ra fico de titu lo s de ac ido g ra so lib re (F F A ) y de e s te r m e tilico de ac ido graso (F A M E ) en cu ltivo s de ce lu la s de E. coli M G 1655 AfadE tra n s fo rm a d a s con pA C Y C que con tien en los h o m o logo s de 'tesA de E. coli (E co lA ), Pectobacterium atrosepticum (P a trA ), Pseudomonas putida (P pu tA ), Vibrio harveyi (V ha rA ), Photorhabdus luminescens (P lum A ), o con pA C Y C que no co n tie n e in se rto (N eg).

Figure 59 (FIG. 59) shows a sequence of nuances encoding a 'T is A of E. coli (SEQ ID NO: 32). Figure 60 (FIG. 60) is a graph of the title of acidic liberty (FFA) and of this type of fatty acid (FAME) in quantitative terms. E. coli MG 1655 AfadE tra ns fo rm adas with pA CYC which have in the homologues of 'tesA de E. coli (E co lA), Pectobacterium atrosepticum (P a trA), Pseudomonas putida (P pu tA), Vibrio harveyi (V ha rA), Photorhabdus luminescens (P lum A), or with pC CYC that does not belong in serto (N eg).

La Figura 61 (FIG. 61) es un g ra fico de titu lo s de F F A y FA M E en cu ltivo s de ce lu la s de E. coli M G 1655 AfadE que sob re e xp re sa n fadD y 'tesA de E. coli (E co li), Pectobacterium atrosepticum (P atr), Photorhabdus luminescens (P lum ), Photobacterium profundum (P pro ), Vibrio harveyi (V hA ), Pseudomonas putida (P pu t), o sin 'tesA (N eg). (Los da tos m arca do s con un a s te risco (*) son de un e xp e rim e n to d is tin to ).

Figure 61 (FIG. 61) is a graph of the titles of FFA and FA ME in the quantitative figure of E. coli MG 1655 AfadE which is based on the presence of FADD and 'tesA of E. coli (E co li), Pectobacterium atrosepticum (P atr), Photorhabdus luminescens (P lum), Photobacterium profundum (P pro), Vibrio harveyi (V hA), Pseudomonas putida (P pu t), or without 'tesA (N eg). (The most important data with a cracked ace (*) are of an e r rim in d d tin tin).

La Figura 62 (FIG. 62) es un g ra fico de titu lo s de F F A y F A M E en cu ltivo s de E. coli M G 1655 AfadE que exp resa n 'tesA de E. co li de t ip o s ilve s tre (TS), el m u tan te S 10 C (S 10C ), o sin 'tesA (N eg).

Figure 62 (FIG. 62) is a graph of FFA and FAME titles in quantitative E. coli MG 1655 AfadE that is based on E. co li de t ip os ilve s tre (TS), the mutant S 10 C (S 10C), or without 'tesA (N eg).

La Figura 63 (FIG. 63) es un g ra fico de p ro du cc ion de FA M E fre n te al t ie m p o de un c ic lo de fe rm e n ta c io n con ce lu la s h o sp e d a d o ra s re co m b in a n te s que exp re sa n tio e s te ra s a en au se n c ia de e s te r s in ta sa exogena .

Figure 63 (FIG. 63) is a graph of the performance of FA ME fre n te at the time of a faith cycle in relation to the impact s re co mb in an te s that exp re sa n unio is te ra sa in au se nc ia of this s ex rs in ta sa exogenous.

La Figura 64 (FIG. 64) es un g ra fico de p ro du cc ion de F FA fren te al t ie m p o de un c ic lo de fe rm e n ta c io n con ce lu la s h o sp e d a d o ra s re co m b in a n te s que exp re sa n t io e s te ra sa en au se n c ia de e s te r s in ta sa exogena .

Figure 64 (FIG. 64) is a graph of F r du ction of F FA at the time of a faith cycle at which time the shooter is issued. Re co mb in an te s that exp re sa nt io is te ra sa in au se nc ia of this you are exogenous.

Descripcion detallada de la invencion

Detailed description of the invention

S a lvo que se de finan de otra m anera , to d o s los te rm in o s te cn ico s y c ie n tificos u tilizados en el p resen te d o cum e n to tie n e n el m ism o s ig n ifica d o que el que e n tie n d e co m u n m e n te un exp e rto h a b itu a l en la m a te ria a la cua l p e rte ne ce la p re sen te invenc ion . A u n q u e en la p ra c tica o e n sayo de la p re sen te invenc ion , pu ed en u tiliza rse m e to d o s y m a te ria le s s im ila re s o e q u iva le n te s a los d e sc rito s en el p re sen te do cu m e n to , a con tinu ac io n se de sc rib en m e to d o s y m a te ria le s ad ecu ad os . En caso de con flic to , la p re sen te m e m o ria de scrip tiva , in c lu ye n d o las d e fin ic ion es , p re va lece ra . A d e m a s , los m a te ria les , m e to d o s y e je m p lo s son so la m e n te ilu s tra tivo s y no p re te nd en s e r lim itan tes.

Unless it is financed in a different way, all the technical and syphilic techniques used in the plan have the same meaning as the one In the meantime, an expert in the fourth quarter of the year in which the invention is invented. Even in the practice of the invention, I can make use of it and I would like to give you the idea of those of the ritual in the country. It is important to me that you will continue to explore in the past two years. In the event of conflict, the scrip tive m e m o ria of scrip tiva, in c lu ye n d o the end of the term, p r va va race ra. On the other hand, the most important, the most important and the most important are the most important and not limited to the limitations.

O tra s ca ra c te ris tic a s y v e n ta ja s de la in ve n c io n se ran obv ias a p a rtir de la s ig u ie n te d e sc rip c io n de ta lla d a y a pa rtir de las re iv ind ica c ion es .

It will be obvious that they will become obvious from the signpost of sc rip c tion of this day. indications .

Definiciones:Definitions:

A lo la rgo de la m em o ria d e sc rip tiva , se pu ede h a ce r una re fe re n c ia u tiliza n d o una a b re v ia tu ra de un no m bre de gen o un no m bre de po lip ep tido , pero se en tie n d e que d icho nom bre a b re v ia d o de gen o p o lip e p tid o re p re se n ta el gene ro de ge ne s o po lip ep tido s , re sp e c tiva m e n te . D ichos nom bres de genes in c luyen to d o s los genes que cod ifica n el m ism o p o lip e p tid o y p o lip e p tid o s ho m o lo g o s que tie n e n la m ism a fun c ion fis io log ica . Los no m bre s de los po lip ep tido s in c luye n to d o s los p o lip e p tid o s que tie n e n la m ism a a c tiv id a d (p o r e jem p lo , que ca ta liza n la m ism a re a cc io n q u im ica fu n dam e n ta l).

With regard to the science of science, it is possible to make a refec tive re u nce or use an opening of a gene number or a number of It can be understood, but it is understood that the name of the gene has been expressed in the genus of genetics, but rather the lipid epitome, re sp ec Tiva Men. These gene names include two genes that encode the same type of lipid, and that they have the same physiological function. . The names of the epiphytic lipids in which the two lipids that have the same type of identity (for the most part, which is the most important in the world) re a tio n c im ica fu n dam in ta l).

A m en os que se in d iq ue o tra cosa , los nu m ero s de reg is tro a los que se hace re fe re n c ia en el p re sen te docum e n to , p roceden de la base de d a to s de l N C BI (National Center for Biotechnology Information, C e n tro N a c ion a l de In fo rm a c ion B io te cn o lo g ica ) m a n te n id a po r el Ins titu to N a c ion a l de S a lud , EE. UU. A m enos que se in d iq ue otra cosa, los nu m ero s de reg is tro son los que figu ran en la base de da tos a m arzo de 2008.

In addition to what is being done, the registration numbers to which reference is made in the document in question, are based on the basis of the To the NC BI ( National Center for Biotechnology Information, C in tro N ac ion to that of In fo rm ac ion B io te cn o lo gica), it is maintained by the Institution at from S to lud, USA UU. Unless otherwise stated, the registration numbers are those that are at the base of the 2008 amzo data.

Los n u m e ro s EC (Enzyme Commission, C o m is io n de E n z im a s ) son los e s ta b le c id o s p o r el C o m ite de N o m e n c la tu ra de la U n ion In te rn ac io na l de B io q u im ica y B io lo g ia M o le cu la r (N C -IU B M B ) (d isp o n ib le en h ttp ://w w w .c h e m .q m u l/a c /u k /iu b m b /e n z y m e /). Los num eros EC a los que se hace re fe re nc ia en el p re sen te do cum e n to , p roceden de la base de da tos de ligan do s de la K E G G , m a n te n id a po r la Kyoto Encyclopedia of Genes and Genomes (E n c ic lo p e d ia de G enes y G e n o m a s de K io to ), p a tro c in ada en pa rte po r la U n ive rs ida d de Tokio . A m en os que se in d iq ue o tra cosa, los nu m e ro s de C E son los que figu ran en la base de da tos a m arzo de 2008.

The EC Numbers ( Enzyme Commission, C om is io n of E nz im as) are the statues by the C om ite de N omenc la tu ra of the U nion In te rn ac io na l of B io chemica and B io lo g ia M o le cu la r (NC -IU BMB) (d isp on ib le in h ttp: // www .chem .qmul / ac / uk / iu bmb / enzyme /). The EC numbers to which reference is made in the P r sente do cum en to, belong to the KEGG league data base, maintained by the Kyoto Encyclopedia of Genes and Genomes (E nc ic I asked for G enes y G enomas de K io to), part of the U n ive rs rs out of Tokyo. In addition to the fact that the EC numbers are those that are at the base of the 2008 amzo data.

En el p re se n te do cu m e n to , los a rticu lo s "un", "u n o " y "u na ", se u tilizan pa ra re fe rirse a un o (a ) o a m as de un o (a ) (es decir, a al m en os uno(a )) de l ob je to g ra m a tica l de l articu lo . C o m o e jem p lo , "un e le m e n to " s ig n ifica un e lem e n to o mas de un elemento.

In this case, the articles "one", "one" and "one" are used to refer to one or more than one ( a) (that is, to one in one) of the object of the article of the article. Like jem p lo, "an e le men to" means an e lem in all More than one item.

En el presente documento, el termino "aproximadamente" significa un valor de ± 20 % de un valor numerico dado.

In this document, the term "approximately" means a value of ± 20 % of a given numerical value.

Por lo tanto, "aproximadamente 60 %" se refiere a un valor de 60 ± (20 % de 60) (es decir, entre 48 y 70).

Therefore, "approximately 60%" refers to a value of 60 ± (20% of 60) (that is, between 48 and 70).

Tal como se usa en el presente documento, la expresion "alcohol deshidrogenasa" (EC 1.1.1. *) es un polipeptido

As used herein, the term "alcohol dehydrogenase" (EC 1.1.1. *) Is a polypeptide

capaz de catalizar la conversion de un aldehido graso en un alcohol (por ejemplo, un alcohol graso). Adicionalmente,

capable of catalyzing the conversion of a fatty aldehyde into an alcohol (for example, a fatty alcohol). Additionally,

un experto habitual en la materia apreciara que algunas alcohol deshidrogenasas tambien catalizaran otras

one of ordinary skill in the art will appreciate that some alcohol dehydrogenases will also catalyze others

reacciones. Por ejemplo, algunas alcohol deshidrogenasas aceptaran otros sustratos ademas de los aldehidos

reactions. For example, some alcohol dehydrogenases will accept other substrates in addition to aldehydes.

grasos. Por lo tanto, dichas alcohol deshidrogenasas inespecificas, tambien se incluyen en esta definicion. En la

fatty Therefore, said non-specific alcohol dehydrogenases are also included in this definition. In the

tecnica se conocen secuencias de polinucleotidos que codifican alcohol deshidrogenasas, y dichas deshidrogenasas

technique known polynucleotide sequences encoding alcohol dehydrogenases, and said dehydrogenases

estan disponibles al publico.

They are available to the public.

La expresion "propiedad alterada" se refiere a una modificacion en una o mas propiedades de un polinucleotido

The term "altered property" refers to a modification in one or more properties of a polynucleotide

mutante o proteina mutante con referencia a un polinucleotido precursor o a una proteina precursora.

mutant or mutant protein with reference to a precursor polynucleotide or a precursor protein.

Ventajosamente, las propiedades que pueden alterarse con respecto a las proteinas fabricadas segun la presente

Advantageously, the properties that can be altered with respect to the proteins manufactured according to the present

divulgacion, incluyen estabilidad oxidativa, especificidad de sustrato, selectividad de sustrato, actividad catalitica,

disclosure, include oxidative stability, substrate specificity, substrate selectivity, catalytic activity,

estabilidad termica, estabilidad del pH, perfil de actividad del pH, resistencia a la degradacion proteolitica, K^m , k^{c a t},

thermal stability, pH stability, pH activity profile, resistance to proteolytic degradation, K ^m , k ^cat ,

relacion entre k^{ca t}/k^m , plegamiento de proteina, induccion de una respuesta inmunitaria, capacidad de unirse a un

relationship between k ^{ca t} / k ^m , protein folding, induction of an immune response, ability to bind to a

ligando, capacidad de unirse a un receptor, capacidad de secretarse, capacidad de translocarse de una manera

ligand, ability to bind to a receptor, ability to secrete, ability to translocate in a way

activa en una membrana, capacidad para mostrase en la superficie de una celula, capacidad de oligomerizar,

active in a membrane, ability to show on the surface of a cell, ability to oligomerize,

capacidad de senalizar, capacidad de estimular la proliferacion celular, capacidad de inhibir la proliferacion celular,

ability to signal, ability to stimulate cell proliferation, ability to inhibit cell proliferation,

capacidad de inducir la apoptosis, capacidad de modificarse por fosforilacion o glicosilacion, capacidad para tratar

ability to induce apoptosis, ability to modify by phosphorylation or glycosylation, ability to treat

enfermedades. En un aspecto, se proporcionan tioesterasas mutantes que proceden de una tioesterasa precursora,

diseases. In one aspect, mutant thiosterases are provided that are derived from a precursor thioesterase,

en donde la tioesterasa mutante tiene al menos una propiedad alterada ya sea in vitro o in vivo, en comparacion con

wherein the mutant thioesterase has at least one altered property either in vitro or in vivo, compared to

las propiedades de la tioesterasa precursora. En un aspecto, la propiedad alterada puede ser una propiedad

the properties of the precursor thioesterase. In one aspect, the altered property can be a property

biofisica, tal como estabilidad termica (punto de fusion T^m ), estabilidad del solvente, estabilidad del soluto,

biophysics, such as thermal stability (melting point T ^m ), solvent stability, solute stability,

estabilidad oxidativa, lipofilia, hidrofilia, estructura cuaternaria, momento dipolar, o punto isoelectrico. En un aspecto,

oxidative stability, lipophilicity, hydrophilicity, quaternary structure, dipole moment, or isoelectric point. In one aspect,

la propiedad alterada puede ser una propiedad bioquimica tal como pH optimo, temperatura optima, fuerza ionica

the altered property can be a biochemical property such as optimal pH, optimal temperature, ionic strength

optima y/o un parametro catalitico enzimatico (tal como, por ejemplo, distribucion del producto, rendimiento

optimal and / or an enzymatic catalytic parameter (such as, for example, product distribution, yield

proporcional o porcentual del producto, actividad especifica, preferencia de sustrato, afinidad de sustrato, inhibicion

proportional or percentage of the product, specific activity, substrate preference, substrate affinity, inhibition

de sustrato, afinidad por el producto, velocidad de recambio, inhibicion del producto, mecanismo cinetico, K^m , k^{c a t},

substrate, product affinity, replacement rate, product inhibition, kinetic mechanism, K ^m , k ^cat ,

k^cat/K^m y/o V^{m e x}). En un aspecto, la propiedad alterada es un cambio de preferencia para sustratos particulares, como

k ^cat / K ^m and / or V ^{m e x} ). In one aspect, the altered property is a change of preference for particular substrates, such as

se refleja, por ejemplo, en un cambio de preferencia para la alcoholisis o la hidrolisis, sustratos de acil CoA o acil

it is reflected, for example, in a change of preference for alcohololysis or hydrolysis, acyl CoA or acyl substrates

proteina transportadora de acilo, sustratos de ester o tioester, sustratos saturados o insaturados, posicion de

acyl transporter protein, ester or thioester substrates, saturated or unsaturated substrates, position of

insaturaciones, especificidad amplia o estrecha (por ejemplo, la capacidad de catalizar una variedad de sustratos o

unsaturations, broad or narrow specificity (for example, the ability to catalyze a variety of substrates or

solo sustratos de una longitud cadena de carbono especifica). En un aspecto, la propiedad alterada puede ser un

only substrates of a specific carbon chain length). In one aspect, the altered property may be a

aumento de preferencia o actividad para sustratos ramificados, sustratos que tienen una posicion de ramificacion

increased preference or activity for branched substrates, substrates that have a branching position

especifica, sustratos de hidroxiacilo, sustratos de cetoacilo, sustratos que dan como resultado un producto que tiene

specific, hydroxyacyl substrates, ketoacyl substrates, substrates that result in a product that has

atributos de combustible deseables (es decir, numero de cetano, indice de octano, estabilidad oxidativa, lubricidad,

desirable fuel attributes (i.e., cetane number, octane number, oxidative stability, lubricity,

punto de ignicion, viscosidad, punto de ebullicion, punto de fusion, punto de fluidez, punto de enturbamiento, punto

flash point, viscosity, boiling point, melting point, pour point, cloud point, point

de obstruccion del filtro en frio, caracteristicas de flujo en frio, aromaticidad, y/o numero de yodo). Las propiedades

of cold filter obstruction, cold flow characteristics, aromaticity, and / or iodine number). The properties

alteradas tambien incluyen una disminucion en la actividad o atenuacion de la hidrolisis del ester, tal como la

altered also include a decrease in activity or attenuation of ester hydrolysis, such as the

hidrolisis de las moleculas del producto deseado, o una disminucion en la toxicidad de la proteina para la celula y/o

hydrolysis of the molecules of the desired product, or a decrease in the toxicity of the protein to the cell and / or

un cambio en el nivel de expresion de la proteina en la celula. En un aspecto particular, la al menos una propiedad

a change in the level of protein expression in the cell. In a particular aspect, the at least one property

alterada es, por ejemplo, un cambio en la capacidad de la tioesterasa para catalizar directa o indirectamente la

altered is, for example, a change in the ability of thioesterase to directly or indirectly catalyze the

sintesis de acil esteres grasos, in vivo o in vitro, tal como mediante transesterificacion.

synthesis of fatty acyl esters, in vivo or in vitro, such as by transesterification.

Tal como se usa en el presente documento, una "secuencia analoga" es una en la que la funcion del gen es

As used herein, an "analog sequence" is one in which the function of the gene is

esencialmente la misma que la de un gen de referencia, tal como, por ejemplo, un gen 'tesA de E. coli.

essentially the same as that of a reference gene, such as, for example, a 'tesA gene of E. coli.

Adicionalmente, los genes analogos incluyen al menos aproximadamente 20 %, por ejemplo, al menos

Additionally, analogous genes include at least about 20%, for example, at least

aproximadamente 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 91 % 92 %, 93 %, 94 %, 95 %, 96 %, 97 %, 98 %, 99 % o 100 % de identidad de secuencia con la secuencia de un gen o

approximately 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91% 92%, 93% , 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the sequence of a gene or

polinucleotido de referencia tal como, por ejemplo, la secuencia de polinucleotidos o de polipeptidos de un gen 'tesA

reference polynucleotide such as, for example, the polynucleotide or polypeptide sequence of a 'tesA gene

o de una tioesterasa 'TesA, respectivamente. En aspectos adicionales, a la secuencia se la aplica mas de una de las

or of a thioesterase 'TesA, respectively. In additional aspects, the sequence is applied more than one of the

propiedades anteriores. Las secuencias analogas se determinan mediante metodos de alineamiento de secuencias

previous properties Analog sequences are determined by sequence alignment methods.

conocidos.

known.

El termino "alineamiento" se refiere a un metodo para comparar dos o mas secuencias de polinucleotidos o de

The term "alignment" refers to a method for comparing two or more polynucleotide or sequence sequences.

polipeptidos con el fin de determinar su relacion entre si. Normalmente, los alineamientos se realizan mediante

polypeptides in order to determine their relationship with each other. Normally, alignments are made by

programas informaticos que aplican diversos algoritmos, sin embargo, tambien es posible realizar un alineamiento

computer programs that apply various algorithms, however, it is also possible to perform an alignment

manualmente. Normalmente, los programas de alineamiento se repiten a traves de posibles alineamientos de

manually. Normally, alignment programs are repeated through possible alignments of

secuencias y puntuan los alineamientos utilizando tablas de sustitucion, empleando una variedad de estrategias para

sequences and score alignments using substitution tables, using a variety of strategies to

alcanzar una posible puntuacion de alineamiento optimo. Como algoritmos de alineamiento habitualmente utilizados

achieve a possible optimal alignment score. As commonly used alignment algorithms

se incluyen, pero sin limitacion, CLUSTALW, (vease, Thompson J.D., Higgins D.G., Gibson T.J., CLUSTAL W:

including, but not limited to, CLUSTALW, (see, Thompson J.D., Higgins D.G., Gibson T.J., CLUSTAL W:

improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific

improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific

gap pe na lties and w e ig h t m a trix cho ice , N u c le ic A c id s R esea rch 22 :4673 -4680 , 1994); C L U S T A LV , (vease , Larkin M .A ., e t al., C L U S T A LW 2, C lu s ta lW y C lu s ta lX ve rs io n 2, B io in fo rm a tics 23 (21 ): 2947 -2948 , 2007 ); Jo tu n -H e in , M usc le e t al., M U S C LE : a m u ltip le seq ue nce a lig n m e n t m e thod w ith reduced tim e and space com p le x ity , BM C B io in fo rm a tics 5: 113, 2004 ); M afft, K a lign , P rob C o ns, y T -C o ffe e (vea se N o tre d a m e e t al., T -C o ffe e : A no ve l m ethod fo r m u ltip le seq ue nce a lign m en ts , Jo u rn a l o f M o le cu la r B io lo gy 302: 205 -217 , 2000). Los p ro g ra m as e je m p la re s que im p le m e n ta n uno o m as de los a lgo ritm o s a n te rio res inc luyen , pero sin lim itac ion , M egA lign de D N A S ta r (D N A S ta r, Inc. 3801 R e g e n t St. M ad ison , W I 53705), M U S C LE , T -C o ffee , C L U S T A LX , C L U S T A LV , Ja lV iew , P hylip , y D iscove ry S tud io de A cce lrys (A cce lrys , Inc., 10188 T e le s is Ct, S u ite 100, S an D iego , C A 92121). En un e jem p lo no lim itan te , se u tiliza M eg A lig n para im p le m e n ta r el a lg o ritm o de a lin e a m ie n to C L U S T A L W con los s ig u ie n te s pa ram e tros: p e n a liza c io n po r hu eco 10, p e n a liza c io n po r lo ng itud de hu eco 0,20, re ta rd o de s e cu e n c ia s d ive rg e n te s (30 % ), peso de tra n s ic io n de A D N 0,50, peso de p ro te ina de la m a triz de la se rie G onne t, peso de A D N de la m a triz IUB.

gap pe na lties and w e ig h t m a trix cho ice, N u c le ic A c id s R esea rch 22: 4673-4680, 1994); CLUSTA LV, (see, Larkin M .A., Et al., CLUSTA LW 2, C lu s ta lW and C lu s ta lX ve rs io n 2, B io in fo rm a tics 23 (21): 2947 -2948, 2007); Jo tu n -H y in, M usc le et al., MUSC LE: amu ltip le seq ue nce a lig nmentme thod w ith reduced tim e and space com p le x ity, BM CB io in fo rm a tics 5 : 113, 2004); M afft, K a lign, P rob C or ns, and T-C or ffe e (see N o tre dameet al., T-C or ffe e: A no see lm ethod fo rmu ltip se se se a a lign m in ts, Jo u rn alof M o le cu la r B io lo and 302: 205-217, 2000). P ro g ra m as e je mp la re s that im p le ment n one or more of the same rhythm osan rio res inc lujan, but without limitation, M egA lign of DNAS ta r (DNAS ta r, Inc. 3801 R egent St. M ad ison, WI 53705), MUSC LE, T-C or ffee, CLUSTA LX, CLUSTA LV, Ja lV iew, P hylip, and D iscove r and S tud io de A cce lrys (A cce lrys, Inc., 10188 T e le s is Ct, S u ite 100, S an D iego, CA 92121). In an e jem p what is not limited, M eg A lig n is used to im p le r the l e r e rm of a lin eam ie n to CLUSTALW with the s ig u ie n s pa pa ram e tros: penalty liza c io n po r hu eco 10, penalty liza c io n po r lo ng itud de hu eco 0.20, re se rd o de se cu en ia sd ive rg in te s (30% ), weight of DNA transfer 0.50, weight of weight of the matrix of the water series, weight of DNA of the IUB matrix.

El te rm in o "a n ticu e rp o s " se re fie re a in m un og lob u linas . Los an ticu e rp o s inc luyen , pero sin lim itac ion , in m u n o g lo b u lin a s o b te n id a s d ire c ta m e n te de cu a lq u ie r esp ec ie de la cua l es d e se a b le p ro d u c ir an ticue rpos . A dem as, en la p re sen te d ivu lg a c io n se in c luye n an ticu e rp o s m od ifica dos . El te rm in o ta m b ie n se re fie re a fra g m e n to s de a n ticu e rp o s que con se rva n la ca p ac idad de un irse al m ism o e p ito po al que ta m b ie n se une el an ticu e rp o in tac to , e in c luye a n ticu e rp o s po lic lon a le s , a n ticu e rp o s m on o c lo n a le s , a n ticu e rp o s q u im e ricos , a n ticu e rp o s a n tiid io tip ico s (a n ti­ ID). Los fra g m e n to s de an ticu e rp o s inc luyen , pero sin lim itac ion , reg io nes d e te rm in a n te s de la co m p le m e n ta rie d a d (C D R , del ing les complementarity-determining regions), reg io nes va r ia b le s de fra g m e n to s m o n o ca te n a rio s (scFv, single chain Fv), reg iones v a ria b le s de cadena pesada (VH ), reg iones v a ria b le s de cadena lige ra (VL). En la p re se n te d ivu lg a c io n ta m b ie n se in c luye n a n ticu e rp o s p o lic lo n a le s y m on oc lona le s . P re fe re n te m e n te , los an ticu e rp o s son a n ticu e rp o s m on oc lona le s .

The rm in or "an ticu e rp os" tea re lo re re in m un og lob u linas. The incumbents include, but without limitation, in munog lo bu lin aso n te asd ire c ta men te de l a lq u ie r esp ie ie from which he is of ro duc go an ticue rpos. In addition, in the case of the report, there is an indication of two reasons. The rm in or ta mb ie tea refers to the year-to-day fragances that underlie the capacity of going to the same place as the one to which unites the year or year, and in which there is a lot of time, even though it is not only right now, but also people who are rich, still e rp osan tiid io typical s (an ti ID). Announcement sentences include, but are not limited to, regions of international law (CDR, English complementarity-determining regions) , regions will vary from fra gmen to smono ca te na rio s (scFv, single chain Fv), heavy chain (VH) regions, various regions from light chain (VL). In the p r se n te d ivu lg ac io n ta mb ie n in c luye nan ticu e rp ospo lic lo na le sym on oc canvas le s. P re fe re n te men te, the years are the same as the osm on the canvas.

El te rm in o "a te n u a r" s ig n ifica de b ilita r, re d u c ir o d ism inu ir. En un e jem p lo , la s e n s ib ilid a d de una e n z im a p a rticu la r a la in h ib ic ion po r re tro a lim e n ta c io n o in h ib ic ion ca u sa d a po r una co m p o s ic io n que no es un p ro du c to o un reactivo (re tro a lim e n ta c io n no e s p e c ific a de la ru ta ) se red uce de ta l m an e ra que la a c tiv id a d de la e n z im a no se ve a fe c ta d a po r la p re sen c ia de un com p ue s to . En un e je m p lo pa rticu la r, la e xp res ion de l gen fabH es se n s ib le a la te m p e ra tu ra y su secu en c ia puede a lte ra rse para d ism in u ir la sen s ib ilidad a las flu c tu a c io n e s de la te m p e ra tu ra . A d e m a s , la e xp res ion de l gen fabH puede a te n u a rse cua nd o se desea o b te n e r a m in o a c id o s ram ificado s . En o tro e jem p lo , una e n z im a que se ha m od ifica do para que sea m en os activa , puede d e c irse que esta a tenuada . P ara a te n u a r la e xp res ion de una en z im a puede u tiliza rse una m o d ifica c io n fu n c io n a l de la secu en c ia que co d ifica una enz im a . Las m o d ifica c io n e s de se cu e n c ia s pueden inc lu ir, po r e jem p lo , una m u tac ion , de lec ion , o in se rc ion de uno o m as nu c le o tid o s en una secu en c ia de genes o una secu en c ia que con tro la la tra n sc rip c io n o la tra d u cc io n de una se cu e n c ia de genes, cuya m o d ifica c io n da com o re su lta d o una reducc ion o in h ib ic ion de la p ro du cc ion de l p roducto gen ico , o hace que el p ro du c to g e n ico no sea fun c ion a l. P o r e jem p lo , la sup res ion fu n c io n a l de fabR en E. coli reduce la rep res ion de la ruta b ios in te tica de ac idos g rasos y pe rm ite a E. coli p ro d u c ir m as ac idos g rasos in sa tu ra d o s (U FA , de l ing les unsaturated fatty acids). En a lgu nos casos, una de le c ion fu n c io n a l se de sc rib e com o una m u tac ion po r in ac tiva c ion (knock-out). P ara a te n u a r la e xp res ion de una e n z im a se d ispone de o tros m etodos. P o r e jem p lo , la a te nua c io n puede rea liza rse m od ifica n d o la secu en c ia que cod ifica el gen com o el descrito a n te rio rm e n te ; c o lo c a n d o el gen ba jo el c o n tro l de un p ro m o to r m en os activo , e x p re s a n d o los A R N in te rfe ren te s , las riboz im as, o las s e cu e n c ia s a n tise n tid o que se d irigen al gen de in te res; ca m b ia n d o el en to rno fis ico o qu im ico , ta l com o la tem p e ra tu ra , el pH, o la co n ce n tra c io n de so lu to , de ta l m an era que la ac tiv idad op tim a del gen o del p ro du c to gen ico no se rea lice ; o a tra ve s de cu a lq u ie r o tra te cn ica c o n o c id a en la m ateria .

The tea rm in o "a te nuar" is the only sign of b ilita r, re duc ir od ism inu ir. In one example, the sensitivity of an immuno-apathetic enzyme in h ib ic ion p r a tro in lim o in h ib ic ion ca u sa da po r a co Because it is not a product or a reagent (but not limited to the specific route), it is assumed that the activity is of the enzyme is not seen at the expense of the power of a community. In an example, the r r r, the fa x H e rp of the gen is that it is only for your time and its sequencing may be allowed to go to the country. It is important to the flow of your actions. In addition, the expression of the FabH gene can be achieved when it is desired to obtain branched branches. In other cases, an enzyme that has been modified to be more active can be said to be tended. In order to continue the evolution of an en z im a can be used a modification of the function of the section that encodes an enzyme. Modifications of signs that can be included, for example, a mu- tion, of lesson, or in section of one or more numbers in a section In the case of genes or a section that, with the work of the science, not the work of a gene sequence, whose modification varies with its number Doing a reduction or reduction in the production of the gene product, or does not make the function of the gene product func- tion. P o jem p lo, the sup p ional end of fabR in E. coli reduces the response of the b ios in tica route of ordinary acids and pe rm ite to E. coli p ro duc ir more common acids in sa tu ra dos (U FA, from unsaturated fatty acids). In some cases, one of the func- tional lesions will be sc rib e com or a positive mutation in knock-out. In order to determine the effect of an enzyme, it has two methods. P er jem p lo, the year can still be done by measuring the sequence coding for the common gene or the one described above. When the gene is lowered with the control of an active agent, the ARNs in re rfe ren te s, the riboz im as, or the san tise n tid o that they go to the gene of interest; It is also possible that the physical or chemical environment, such as the temperature, the pH, or the relationship between the other, so it was that the optimal activity of the gene or of the generative gene is not related; or through which I know what I know in the matter.

El te rm in o "b io c ru d o " se re fie re a un b io co m b u s tib le que puede u tiliza rse com o su s titu to de co m b u s tib le s ba sad os en pe tro leo . A de m as , el b iocrudo , com o el pe tro leo crudo , puede tra n s fo rm a rs e en o tros com b us tib les , po r e jem p lo , gaso lina , d iese l, com b u s tib le de av iac io n o ga so il para ca le facc ion . A d e m a s , el b iocrudo , com o el pe tro leo crudo, puede tra n s fo rm a rs e en o tros p ro du c to s qu im icos de u tilidad in du s tria l para su uso, po r e jem p lo , en p roductos fa rm a ce u tico s , cosm e ticos , b ienes de con sum o , p ro ceso s in du s tria le s , etc. Una com p o s ic io n de b ioc ru do puede co m p re n d e r, po r e jem p lo , h id ro ca rb u ro s , p ro d u c to s de h id ro ca rbu ro , es te re s de ac ido s g ra sos y /o ce to n a s a lifa ticas , o una com b in ac io n de los m ism os. En un a sp e c to p re fe rido , una com p o s ic io n de b ioc ru do es ta co m p u e s ta por h id ro ca rbu ros , po r e jem p lo , h id ro ca rb u ro s a lifa tico s (p o r e jem p lo , a lcanos, a lqu eno s , a lq u in o s ) o a ro m atico s .

The tea rm in o "b io c ru d o" refers to a commonplace that can be used as its co-operative status in the future. In addition, the bureaucratic, like the crude oil, can be used in other countries, for example, for gaso lina, for that, for the tib bus He told her that he didn't want to do it. In addition, the bureaucratic one, such as crude oil, can be used in other chemical ways to use it for use, because re jem p lo, in cosmetic products, cosmetics, consumer goods, for example in du s tria le s, etc. A com io nio of b ioc ru do can co mp re nder, po re jem p lo, h id ro ca rb u ro s, p ro duc to s de h id ro ca rbu ro, this re s of Here you are and / or ceda nasa lina ticas, or a com b in ation of the same. In an important context, a biocomputer composition is a matter of interest, that is why, what is the reason for life. tico s (by jem p lo, to lcanos, to lqu eno s, to lq u in os) or to m mtico s.

El te rm in o "b io d ie se l" se re fie re a un t ip o p a rticu la r de b io co m b u s tib le que puede u tiliza rse en m o to re s d iese l. El b io d ie se l puede s e r un su s titu to de l d iese l tra d ic io n a l, que n o rm a lm e n te p rocede de l pe tro leo . El b iod iese l puede u tiliza rse en m o to re s d iese l de co m b u s tio n in te rna en fo rm a pura, que se co n o ce com o b iod iese l "n o d ilu ido ", o com o una m ezc la en cu a lq u ie r co n ce n tra c io n con un d iese l basado en pe tro leo . Una com p o s ic io n de b iod iese l tam b ien puede c o m p re n d e r d ive rso s ad itivo s ad ecu ad os . El b iod iese l pued e e s ta r co m p u e s to po r h id ro ca rb u ro s o este res. En un aspec to , el b iod iese l es ta c o m p u e s to po r es te res g rasos , ta le s com o los es te re s m e tilico s de ac ido s grasos (F A M E ) o los es te res e tilico s de ac ido s g ra sos (F A E E ). En un asp ec to p re fe rido , es to s F A M E y F A E E estan co m p u e s to s po r g ru po s de ac ilo g ra so que tie n e n una lo ng itud de ca d e n a de ca rb o n o de a p ro x im a d a m e n te 8-20, 10­ 18 o 12-16. Los es te res g ra sos u tilizad os com o b iod iese l pueden co n te n e r cad en as de ca rbono rectas, ram ificadas, sa tu ra d a s o insa tu radas .

The rm in o "b io d ie se l" refers to a t ip or p a rticu la r de b io co m b u s tib le which can be used in m o to re s d sese l. The b io d ie l can be a substitution of l d iese l tra d ic io n a l, which p r o a lm e n p p you of the peter trophy. The b iod iese l can be used in mo to re sd iese l de co mbus tio n in te rna in pure form, which is known as ob iod ie l "nod ilu gone", or as a It is based on a period of time based on petroleum. A com- bity of b iod iese l also can also be ad dive r s ive s ad itivo s ad ecu ad os. The b iod iese s can r co m p u e s to po r h id ro ca rb u ro s or this res. In one respect, the bimonth is that it is composed of these resources, such as the fatty acids of fatty acid (FAME) or the latter. de ac ido sg ra sos (FAEE). In an asp e r p r r e r r e r, it is all FAME and FAEE are responsible for their acronym so that they have a channel of the name of the channel of ap im adamen te 8-20, 10 18 or 12-16. These are used as a basis that can be straight, branched, in your own way or in your own way.

El te rm in o "b io co m b u s tib le " se re fie re a cu a lq u ie r co m b u s tib le p ro ced en te de b iom asa . La b iom asa es un m a te ria l b io lo g ico que puede tra n s fo rm a rs e en b ioco m bus tib le . Una fu e n te e je m p la r de b iom asa es la m a te ria veg e ta l. P o r ejemplo, como biomasa, puede utilizarse maiz, cana de azucar y cesped de pradera. Otro ejemplo no limitativo de biomasa es la materia animal, por ejemplo, estiercol de vaca. La biomasa tambien incluye productos de desecho de la industria, agricultura, silvicultura, y domesticos. Como ejemplos de dichos productos de desecho se incluyen, sin limitacion, desechos de fermentacion, paja, madera, aguas residuales, basura y restos de comida y glicerol. La biomasa tambien incluye fuentes de carbono, tales como hidratos de carbono (por ejemplo, azucares). Los biocombustibles pueden sustituirse por combustibles basados en petroleo. Por ejemplo, los biocombustibles incluyen combustibles para el transporte (por ejemplo, gasolina, diesel, combustible de aviacion, etc.), combustibles de calefaccion, y combustibles generadores de electricidad. Un biocombustible es una fuente de energia renovable. Como ejemplos no limitativos de biocombustibles se incluyen biodiesel, hidrocarburos (por ejemplo, alcanos, alquenos, alquinos, o hidrocarburos aromaticos), y alcoholes procedentes de biomasa.

The rm in or "b io co mbus tib le" tea refers to what the co lbus tib le p ro ced in b iom asa tea. The search is a logical way that can be carried out by bus or bus. A fu in te e je mp la r of b iom asa is ma ve r e ta l. By For example, as biomass, corn, sugarcane and prairie grass can be used. Another non-limiting example of biomass is animal matter, for example, cow dung. Biomass also includes waste products from industry, agriculture, forestry, and domestic. Examples of such waste products include, without limitation, fermentation waste, straw, wood, sewage, garbage and leftover food and glycerol. Biomass also includes carbon sources, such as carbohydrates (for example, sugars). Biofuels can be replaced by petroleum-based fuels. For example, biofuels include fuels for transportation (for example, gasoline, diesel, aviation fuel, etc.), heating fuels, and electricity generating fuels. A biofuel is a renewable energy source. Non-limiting examples of biofuels include biodiesel, hydrocarbons (for example, alkanes, alkenes, alkynes, or aromatic hydrocarbons), and biomass alcohols.

En el presente documento, la expresion "longitud de la cadena de carbono" se define como el numero de atomos de carbono en una cadena de carbono de un sustrato de tioesterasa o de un derivado de acido graso. La longitud de la cadena de carbono de una molecula particular se indica como Cx, en la que el subindice "x" se refiere al numero de carbonos en la cadena de carbono. Tal como se usa en el presente documento, la expresion "cadena larga" se refiere a aquellas moleculas que tienen una cadena de carbono con una longitud de aproximadamente 15 a aproximadamente 20 carbonos (por ejemplo, C¹⁵ , C¹⁶ , C¹⁷ , C¹⁸ , C¹⁹ o C²⁰ ). La expresion "cadena corta" se refiere a aquellas moleculas que tienen una cadena de carbono con una longitud de aproximadamente 8 a aproximadamente 14 carbonos (por ejemplo, C8, C⁹ , C¹⁰ , C¹¹ o C¹² ).

Here, the term "carbon chain length" is defined as the number of carbon atoms in a carbon chain of a thioesterase substrate or a fatty acid derivative. The carbon chain length of a particular molecule is indicated as Cx, in which the subscript "x" refers to the number of carbons in the carbon chain. As used herein, the term "long chain" refers to those molecules that have a carbon chain with a length of about 15 to about 20 carbons (eg, C ¹⁵ , C ¹⁶ , C ¹⁷ , C ¹⁸ , C ¹⁹ or C ²⁰ ). The term "short chain" refers to those molecules that have a carbon chain with a length of about 8 to about 14 carbons (for example, C8, C ⁹ , C ¹⁰ , C ¹¹ or C ¹² ).

La expresion "fuente de carbono" significa un sustrato o compuesto que es adecuado para utilizar como una fuente de carbono para el crecimiento de celulas procariotas o eucariotas simples. Las fuentes de carbono pueden estar en diversas formas, incluyendo, pero sin limitacion, polimeros, hidratos de carbono, acidos, alcoholes, aldehidos, cetonas, aminoacidos, peptidos, gases (por ejemplo, CO y CO² ), y formas similares. Estas incluyen, por ejemplo, diversos monosacaridos tales como glucosa, fructosa, manosa y galactosa; oligosacaridos tales como fructooligosacaridos y galactooligosacaridos; polisacaridos tales como xilosa y arabinosa; disacaridos tales como sacarosa, maltosa y turanosa; material celulosico tal como metilcelulosa y carboximetilcelulosa sodica; esteres de acidos grasos saturados o insaturados tales como succinato, lactato y acetato; alcoholes tales como metanol, etc., o mezclas de los mismos. La fuente de carbono tambien puede ser un producto de la fotosintesis, incluyendo, pero sin limitacion, la glucosa. El glicerol tambien puede ser una fuente de carbono eficaz. A partir de cualquier cantidad de fuentes naturales y renovables pueden generarse fuentes de carbono adecuadas, incluyendo particularmente biomasa procedente de residuos agricolas, urbanos e industriales, siempre que el material pueda utilizarse como un componente de una fermentacion para proporcionar una fuente de carbono. Las fuentes de biomasa incluyen rastrojos de maiz, cana de azucar, cesped de pradera, materia animal, o materiales residuales.

The term "carbon source" means a substrate or compound that is suitable for use as a carbon source for the growth of simple prokaryotic or eukaryotic cells. Carbon sources may be in various forms, including, but not limited to, polymers, carbohydrates, acids, alcohols, aldehydes, ketones, amino acids, peptides, gases (eg, CO and CO ² ), and similar forms. These include, for example, various monosaccharides such as glucose, fructose, mannose and galactose; oligosaccharides such as fructooligosaccharides and galactooligosaccharides; polysaccharides such as xylose and arabinose; disaccharides such as sucrose, maltose and turanosa; cellulosic material such as methyl cellulose and sodium carboxymethyl cellulose; esters of saturated or unsaturated fatty acids such as succinate, lactate and acetate; alcohols such as methanol, etc., or mixtures thereof. The carbon source can also be a product of photosynthesis, including, but not limited to, glucose. Glycerol can also be an effective carbon source. From any amount of natural and renewable sources suitable carbon sources can be generated, particularly including biomass from agricultural, urban and industrial waste, provided that the material can be used as a component of a fermentation to provide a carbon source. Biomass sources include corn stubble, sugarcane, prairie grass, animal matter, or waste materials.

La expresion "integracion cromosomica" significa el proceso mediante el cual se introduce una secuencia entrante en el cromosoma de una celula hospedadora. Las regiones homologas del ADN transformante se alinean con las regiones homologas del cromosoma. Despues, la secuencia que esta entre las cajas de homologia, puede reemplazarse por la secuencia entrante en un doble cruce (es decir, por recombinacion homologa). En algunos aspectos, las secciones homologas de un segmento cromosomico inactivador de una construccion de ADN se alinean con las regiones homologas flanqueantes de la region cromosomica autoctona del cromosoma microbiano. Posteriormente, la construccion de ADN elimina la region cromosomica autoctona en un doble cruce.

The term "chromosomal integration" means the process by which an incoming sequence is introduced into the chromosome of a host cell. The homologous regions of the transforming DNA align with the homologous regions of the chromosome. Then, the sequence that is between the homology boxes can be replaced by the incoming sequence in a double cross (ie, by homologous recombination). In some aspects, the homologous sections of an inactivating chromosomal segment of a DNA construct align with the flanking homologous regions of the autochthonous chromosome region of the microbial chromosome. Subsequently, DNA construction eliminates the native chromosomal region in a double cross.

La expresion "punto de enturbamiento" se refiere a la temperatura de un liquido a la cual los solidos disueltos ya no son completamente solubles, precipitando como una segunda fase y dando al liquido un aspecto turbio. Esta expresion es relevante para diversas aplicaciones con consecuencias algo o completamente diferentes. En la industria petrolera, el punto de enturbamiento se refiere a la temperatura por debajo de la cual la cera u otros hidrocarburos pesados cristalizan en un petroleo crudo, aceite o combustible refinado, para formar un aspecto turbio. La presencia de cera solidificada influye en el comportamiento de flujo del fluido, aumentando la tendencia a obstruir los filtros de combustible / inyectores y otras piezas de la maquinaria, causando la acumulacion de cera en superficies frias (por ejemplo, en superficies de tuberias o termocambiadores) y cambiando incluso las caracteristicas de emulsion con agua. El punto de enturbamiento es un signo de la tendencia que tiene el aceite a obstruir los filtros o los pequenos orificios a bajas temperaturas de funcionamiento. El punto de enturbamiento de un tensioactivo no ionico o solucion de glicol es la temperatura a la cual la mezcla comienza a separarse en dos o mas fases, volviendose asi turbia. Este comportamiento es caracteristico de los tensioactivos no ionicos que contienen cadenas de polioxietileno, que puede mostrar un comportamiento de solubilidad inversa frente a la temperatura en agua y, por lo tanto, pueden "enturbiarse" en algun momento a medida que aumenta la temperatura. Los glicoles que demuestran este comportamiento se conocen como "glicoles de punto de enturbamiento" y se utilizan como inhibidores de esquisto. Normalmente, la salinidad tambien influye en el punto de enturbamiento, siendo generalmente mas bajo en liquidos mas salinos.

The term "cloud point" refers to the temperature of a liquid at which dissolved solids are no longer completely soluble, precipitating as a second phase and giving the liquid a cloudy appearance. This expression is relevant for various applications with somewhat or completely different consequences. In the oil industry, the cloud point refers to the temperature below which wax or other heavy hydrocarbons crystallize in a crude oil, oil or refined fuel, to form a cloudy appearance. The presence of solidified wax influences the fluid flow behavior, increasing the tendency to clog fuel filters / injectors and other parts of the machinery, causing wax to build up on cold surfaces (for example, in pipe surfaces or heat exchangers ) and even changing the emulsion characteristics with water. The cloud point is a sign of the tendency of the oil to clog the filters or small holes at low operating temperatures. The cloud point of a non-ionic surfactant or glycol solution is the temperature at which the mixture begins to separate into two or more phases, thus becoming cloudy. This behavior is characteristic of non-ionic surfactants that contain polyoxyethylene chains, which can show a behavior of inverse solubility against temperature in water and, therefore, can "get cloudy" at some point as the temperature rises. Glycols that demonstrate this behavior are known as "cloud point glycols" and are used as shale inhibitors. Normally, salinity also influences the cloud point, being generally lower in more saline liquids.

La expresion "aditivo que reduce el punto de enturbamiento" se refiere a un aditivo que puede anadirse a una composicion para disminuir o reducir el punto de enturbamiento de la composicion, como se ha descrito anteriormente.

The term "additive that reduces cloud point" refers to an additive that can be added to a composition to decrease or reduce the cloud point of the composition, as described above.

La expresion "condiciones que permiten la produccion del producto", se refiere a cualquier condicion de fermentacion que pe rm ita a un h o sp e d a d o r de p ro d u cc io n p ro d u c ir un p ro d u c to de sea do , ta l com o a c il-C o A o d e riva d o s de ac ido s g rasos , in c luye nd o , po r e jem p lo , ac ido s g rasos, h id ro ca rbu ros , a lcoh o le s g rasos, ceras o es te res g rasos. Las co n d ic io n e s de fe rm e n ta c io n g e n e ra lm e n te com p ren den m uch os pa ram e tro s . C om o con d ic io n e s e je m p la re s se inc luyen , pero sin lim itac ion , in te rva los de tem p e ra tu ra , n ive les de ox ige nac ion , in te rva los de pH y co m p o s ic io n de los m ed io s (p o r e jem p lo , d iso lve n te s y so lu tos). C ada una de es ta s con d ic ion es , de m an e ra in d iv idu a l y en co m b in ac io n , pe rm ite c re ce r al h o sp e d a d o r de p ro du cc ion . C o m o m ed io s e jem p la re s se inc luyen ca ldo s o ge les. En gene ra l, un m ed io a d e cu a d o in c luye una fu e n te de ca rbono , ta l com o g lucosa , fruc tosa , ce lu losa , o fu e n te s s im ila res, que puede m e ta b o liz a r d ire c ta m e n te el m ic ro o rga n ism o . A de m as , en el m ed io , pueden u tiliza rse e n z im a s para fa c il ita r la m o v iliza c io n (p o r e jem p lo , la d e s p o lim e riz a c io n de l a lm id o n o de la ce lu lo sa a a zu ca re s fe rm e n ta b le s ) y el m e ta b o lism o p o s te r io r de la fu e n te de ca rb on o . P a ra d e te rm in a r si las co n d ic io n e s de c u ltivo son a d e c u a d a s pa ra la p ro du cc ion de l producto , el h o sp e d a d o r de p ro du cc ion puede cu ltiva rse d u ran te a p ro x im a d a m e n te 4, 8, 12, 24, 36, 48 o 72 horas. D u ra n te el cu ltivo o d e sp u e s de l m ism o, las m u e s tra s pueden o b te n e rse y a n a liza rse para d e te rm in a r si las co n d ic io n e s de cu ltivo pe rm iten la p ro du cc ion de l p roducto . P o r e jem p lo , los h o sp e d a d o re s de p ro du cc ion en la m u e s tra o en el m ed io en do n d e se cu ltiva ro n los h o sp e d a d o re s de p ro du cc ion , pu ed en e xa m in a rse pa ra d e te rm in a r la p re sen c ia del p ro du c to deseado . C u an do se exa m in a la p re sen c ia de un producto , pueden u tiliza rse ensayos, ta le s com o, pero sin lim itac ion , T LC , H P LC , G C /F ID , G C /M S , LC /M S , M S, a s i com o los p ro p o rc io n a d o s en los e je m p lo s de este docum e n to .

The expression "conditions that allow the production of the product" refers to any fermentation condition That is only a few years old, but it is only for a few years, as well as two years old, in which there is no or, po re jem p lo, ac se sg rasos, h id ro ca rbu ros, a lcoh or le sg satos, waxes or these ress. The faith rm connections in ta n nnio ra lm en te com p ren den many others. If you need it, you will find it, but without limitation, it will include those of temperature, levels of oxygenation, and those of pH and COMPOSITIONS OF THE MEDIA (for example, only two years ago). Each of these conditions is, from year to month, in common with each other, at the age of the age of service. C omom ed io se jem p la re s inc inc uyen ca ldo so geles. In general, a small group also has a carbon fiber, a small group of fruit, fruitful fruit, a glass of tea, or other images, which can help me liz ard ire c ta men te el m ic ro o rga n ism o. In addition, in the meantime, enzymes can be used to facilitate the movement (for example, the detrimental limitation of the same id from the ce lu lo sa aa zu ca re s fe rm in ta b le s) and the me ta bo lism opposition te r io r from the fu in te r r on. In order to see if the quantitative conditions are adequate for the product, the current age of the product may be quantitative. ro x im adamen te 4, 8, 12, 24, 36, 48 or 72 hours. According to the quantitative statement of the same, the samples can be obtained and can be used to determine whether the quantitative reports pertain to the p ro du cc ion of lp roducto. P o jem p lo, the ho p ered re s of p ro du cc ion in the sample or in the world where the ho ho e r eados p s of p ro du cc ion, pu In this case, you will be able to see the desired price. When testing a product, tests can be used, as well as, but not limited to, T LC, HP LC, GC / F ID, GC / MS, LC / MS, MS, as well as the risks in the examples of this document at all.

Las e xp res ione s "se cu e n c ia co n se n so " o "se cu e n c ia can on ica " se re fie ren a una se cu e n c ia de a m ino ac id os a rq u e tip ica fre n te a la que se com p ara n to d a s las v a ria n te s de una p ro te ina o secu en c ia de in te res pa rticu la r. C u a lq u ie ra de las dos e xp res ione s ta m b ie n se re fie re a una se cu e n c ia que expone los nu c le o tido s que estan p re sen te s con m a yo r fre cu e n c ia en una se cu e n c ia de p o lin u c le o tid o s de in te res. P a ra cad a po s ic ion de una p ro te ina , la secu en c ia con sen so p ro p o rc io n a el am in o a c id o que es m as a b u n da n te en esa pos ic ion en el a lin e a m ie n to de secu en c ias .

The e xp res ions "are known as canon" or "canonical" refer to a sign of a common interest in the typical freq The one that will be bought for the various aspects of a project or section of interest in the country. One of the two leaders who refers to a statement that exposes the nurtures that are present with my mother in one of the lines of interest or interest. For the purpose of a parent, the section with him is only possible in the United States, which is more abundant in that position. ion in the lin eam ie n to secu in c ias.

T a l com o se usa en el p resen te do cu m e n to , la e x p re s io n "m u ta c io n co n se n so " se re fie re a una d ife ren c ia en la s e cu e n c ia de un gen de pa rtida y una se cu e n c ia co n sen so . Las m u ta c io n e s con sen so se id en tifican c o m p a ra n d o las s e cu e n c ia s de l gen de p a rtida y la se cu e n c ia con sen so resu lta n te de un a lin e a m ie n to de secu en c ias . En a lgunos asp ec tos , las m u ta c io n e s c o n s e n s o se in tro d u ce n en el gen de p a rtida de ta l m a n e ra que se v u e lv e m as s im ila r a la se cu e n c ia consenso . Las m u ta c io n e s con sen so ta m b ie n in c luye n cam b io s de a m ino ac id os que cam b ian un a m in o a c id o en un gen de pa rtida po r un am in o a c id o que se e n cu e n tra m as fre c u e n te m e n te en un a lin e a m ie n to de se cu e n c ia s m u ltip le (M S A , multiple sequence alignment) en esa pos ic ion en re lac ion con la fre cu e n c ia de ese a m in o a c id o en el gen de pa rtida . P o r lo tan to , la exp res ion "m u ta c io n co n se n so " se re fie re a cu a lq u ie r cam b io de a m in o a c id o que re e m p la ce un a m in o a c id o de l gen de pa rtida po r un a m in o a c id o que sea m as a b u n d a n te en el M S A que el a m in o a c id o nativo.

As it is used in the present, the exp "sion" mu ta con cio n n n "refers to a period in which the se of a birth gene and one that is common. Mutations with meaning are identified in comparison to those of the birth gene and the sequence with a summary of a year-end report. secu in c ias. In some cases, the many consensus will be introduced into the gene in the same way that is seen in the same way as the consensus. Mutations with sen so ta mb ie n in c luy n cam b io s de am ino ac id os that changed an am in or a c id or in a gene of parrida por r a am in or that I know what is being done more frequently in one year of the most important sequence (MSA, multiple sequence alignment) in that position in re In relation to the frequency of that am in or in the birth gene. For the most part, the expression "mu ta con io n con n so" refers to how much change is made in the country that represents Am in o a c id o of the birth gene for a more in or more abundant in the MSA than the native or native.

Las e x p re s io n e s "su s titu c io n e s co n se rva tiva s " o "s u s titu c io n e s co n se rva d a s " se re fie ren , po r e jem p lo , a sus titu c ion es en las que se rea lizan una o m as de las s ig u ie n tes su s titu c io n e s de am ino ac id os : ree m p la zo de un a m ino ac id o a lifa tico , ta l co m o a lan ina , va lina , le uc in a e iso leuc ina , po r o tro a m in o a c id o a lifa tico ; re e m p la zo de una se rina po r una treon in a ; ree m p la zo de una tre o n in a po r una serina ; ree m p la zo de un resto ac ido , ta l com o ac ido asp a rtico y ac ido g lu tam ico , po r o tro resto ac ido ; ree m p la zo de un resto que lleva un g rupo am ida , ta l com o a sp a ra g in a y g lu tam ina , po r o tro res to que lleva un g ru p o am ida ; in te rca m b io de un res to basico , ta l com o h is tid ina , lis ina y a rg in ina , po r o tro res to bas ico ; y re e m p la zo de un res to a ro m a tico , ta l com o trip to fa n o , fe n ila la n in a y tiro s in a , po r o tro res to a ro m a tico ; o re e m p la zo de a m in o a c id o s pequenos , ta le s com o g lic ina , a lan ina , se rina , tre o n in a y m e tion ina , po r o tro am in o a c id o pequeno. En la te cn ica se conocen su s titu c io n e s de am in o a c id o s que g e n e ra lm e n te no a lte ran la ac tiv ida d e s p e c ific a y las d e sc rib e n , po r e jem p lo , H. N e u ra th y R .L. H ill, en T h e P ro te ins , A c a d e m ic P ress, N u eva Y ork , 1979. Las m o d ifica c io n e s co n se rva tiva s u tiles in c luyen de a lan ina a c is te ina , g lic in a o serina ; de a rg in in a a iso leuc ina , lis ina , m e tio n ina u o rn itina ; de a s p a ra g in a a ac ido aspartico , g lu tam ina , ac ido g lu ta m ico o h is tid ina ; de ac ido asp a rtico a asp a ra g in a , g lu tam ina o ac ido g lu tam ico ; de c is te ina a m e tion ina , se rina o treon in a ; de g lu tam ina a asp a ra g in a , ac ido asp a rtico o ac ido g lu tam ico ; de ac ido g lu tam ico a a sp a ra g in a , ac ido asp a rtico o g la tm ina ; de g lic in a a ac ido aspartico , a lan ina o pro lina ; de h is tid in a a asp a ra g in a o g lu tam ina ; de iso leu c in a a leuc ina , m e tio n ina o va lina ; de le uc in a a iso leu c in a , m e tio n ina o va lina ; de lis ina a a rg in ina , g lu tam ina , ac ido g lu tam ico , iso leuc ina , m e tio n in a u o rn itin a ; de m e tio n in a a c is te ina , iso leu c in a , le uc in a o va lina ; de fe n ila la n in a a h is tid ina , L-D opa, leucina, m e tio n ina , treon in a , trip to fa n o , tiros ina , 3 -fe n ilp ro lin a , 4 -fe n ilp ro lin a o 5 -fen ilp ro lina ; de p ro lin a a ac ido L -1 -tio a zo lid in -4 -ca rb o x ilico o ac ido D- o L -1 -o xa zo lid in -4 -ca rb o x ilico ; de se rin a a c is te ina , m e tio n ina o tre o n in a ; de tre o n in a a m e tio n ina , se rina o va lina ; de tr ip to fa n o a tiro s in a ; de tiro s in a a L -D opa , h is tid ina o fe n ila la n in a ; y de va lin a a iso leuc ina , le uc in a o m e tion ina .

The exp "s" their subtitle co n se rva tiva s "or" their subtitle co n se rva das "is re lo re ren, po re jem p lo, its subtitle is in which one or more of the following substitutions are carried out: replacing the zoo of a lifelong love, such as the moor lan ina , va lina, le uc in ae iso leuc ina, po tro am in o a c id or lifts; re e m p the zo of a country for a threon in a; ree m p the zo of a three to a serine; ree m p the zo of an acidic remainder, such as arctic aspic and gama tamico, by the rest acid; re m p the zo of a remainder that has a large group, such as a group, and a group that carries a large group; In the middle of a basic res, such as the inactive, inactive, and basic inward; and re e m p the zo of a robotic, such as the trip or fate, faith in the n in a and th e r th in a, r o r th r a r a r r a r r a r m a tica; In the small area, small groups, such as those that are inactive, will be ruled, three years, and less than a small number. In the technical field, its substitution of am in or of the languages that generate the short-term activity and those of the sc ribs are known, in particular, H. N eu ra th and R .L. H il, in T he P ro te ins, A cadem ic P ress, N u eva Y ork, 1979. The modifications made useful in c lien de a lan ina ac is te ina , g lic in ao serina; from a rg in in a a iso leuc ina, lis ina, m e tio n ina u o rn itina; from a s p a ra g in a a aspartic acid, g lu tam ina, g lu ta mico or h is tid ina; from arctic asp to asp a ra g in a, g lu tam ina or gama tamico; de c is te ina a m e tion ina, rina or treon in a; from g lu tam ina a asp a ra g in a, arctic asp arc or gamico acid; from a g th tamic a a sp a ra g in a, arctic asp a g o tm ina; from g lic in a aspartic acid, to lan ina or pro lina; from h is tid in a a asp a ra g in a or g lu tam ina; from iso leu c in a a leuc ina, m e tio n ina o va lina; from le uc in a a iso leu c in a, m e tio n ina o va lina; from lis ina a a rg in ina, g lu tam ina, ac i g lu tamico, iso leuc ina, m e tio n in a u o rn itin a; de m e tio n in a a c is te ina, iso leu c in a, le uc in a o valine; of faith n ila la n in aah is tid ina, LD opa, leucine, me uncle ina, treon in a, trip tA © fa no, ina shots, 3 -fe n ilp ro lin a, 4 -fe n ilp ro lin ao 5 -fen ilp ro lina; from p ro lin a a l gone L -1 -tio a zo lid in -4 -ca rb o x ilic or ad gone D- o L -1 -o xa zo lid in -4 -ca rb o x il; from surrender to c is te ina, m e unio n ina o tre o n in a; from three to three years, it is ruined or valine; from tr ip tto fa n o to shot s in a; shooting s in a a L-D opa, h is tid ina or faith n ila la n in a; and from there lin a a le leuc ina, le uc in a or m e tion ina.

La exp res ion "co rre sp o n d e a" se re fie re a un resto de am ino ac id o en una p rim e ra secu en c ia de p ro te inas que, en cua n to a su pos ic ion , es e q u iva le n te a un res to de a m in o a c id o en una seg un da secu en c ia de p ro te inas de re fe renc ia , de b ido que el res to de la p rim e ra secu en c ia de p ro te inas se a linea con el resto de la secu en c ia de p ro te in a s de re fe re nc ia u tiliza n d o te c n ic a s b io in fo rm a tica s , po r e jem p lo , u tiliza n d o los m e to d o s de sc rito s en el p re se n te d o cu m e n to para p re p a ra r un a lin e a m ie n to de se cu e n c ia s . El res to co rre sp o n d ie n te en la p rim e ra se cu e n c ia de p ro te in a s se a s ig n a d e sp u e s al n u m ero de res to en la s e g u n d a s e c u e n c ia de p ro te in a s de re fe re nc ia . La p rim era se cu e n c ia de p ro te inas puede s e r an a loga a la seg un da secu en c ia de p ro te inas o no a n a loga a la seg un da se cu e n c ia de p ro te inas , au nque se p re fie re que las dos s e cu e n c ia s de p ro te in a s sean s e cu e n c ia s ana logas . P or e jem p lo , cua nd o la secu en c ia de a m in o a c id o s de una 'T e sA de E. co li, S E Q ID NO:31 exp ue s ta en la FIG. 58, se u tiliza com o una secu en c ia de re fe renc ia , a cada uno de los res tos de a m in o a c id o s en o tra p ro te ina a line ad a de in te res o en una p ro te ina ana loga , se le puede a s ig n a r un nu m ero de resto c o rre sp o n d ie n te a los num eros de los res tos 1-182 de la S E C ID NO: 31. P o r e jem p lo , en la FIG. 47, las s e cu e n c ia s de a m in o a c id o s a line ad as estan re la c io n a d a s o se c o rre s p o n d e n con la se cu e n c ia de una 'T e sA de E. c o li. P o r c o n s ig u ie n te , a una po s ic io n da da en o tra t io e s te ra s a de in te res, ya sea una tio e s te ra s a p re cu rso ra o m u tan te , se la puede a s ig n a r una posic ion co rre sp o n d ie n te en la secu en c ia 'TesA , u tilizan do te cn ica s b io in fo rm a tica s conoc idas , ta le s com o las desc ritas en el p re sen te docum e n to .

The expiration "cor rre sp wand" refers to a rest of the year in a first-time section that, in terms of its position, It is equitable to a rest of am in or to a second a sec- ond day in re fe ne of re fe rins of faith, so that the rest of the p r e The secu in the p r te ines line up with the rest of the p se te in the p re te n t of re re nc ia or use of the cn ic asb io in fo rm a tics s, po In this regard, he uses the methods of science in which he is responsible for a year in which he is concerned. The answer to the sp e rd e n te in the p rim e ra se cu en de p ro te sins thus sp sp e ns to the number of res in the second sequence of p ro te re fe re nc ia. The first period of time may be analogous to the second one day after the second period or not analogous to the second one day after the second day. Here, even though it is true that the two are aware of each other, they are similar. By In other words, the section in the am in or in the terms of an E. co li, SEQ ID NO: 31 is shown in FIG. 58, it is used as a referable section of faith, to each of the rest of the countries in or through the line of interest or in A person who is still analogous, can be assigned a number of remaining tax spheres to the numbers of rests 1-182 of SEQ ID NO: 31. P ore jem p lo, in FIG. 47, the signs of am in or of idyllic lines are re na e nadaso or correspond with the signal of a T e sA of E. co li. For example, a few days in or out of time, either an uncle is the one who knows how to do it, it can be done. ig as nar one posit ion co rre sp ond ie n t in secu in ai c 'tesa, or tilizan do tea ica cn sb io in fo rm comings s tica conoc, ta le s com or ritas desc in the p re sen do docum en to.

El te rm in o "d e le c io n ", cua nd o se u tiliza en el con tex to de una se cu e n c ia de am ino ac id os , s ig n ifica una de le c ion o e lim in a c io n de un res to de la secu en c ia de am in o a c id o s de una p ro te ina p recu rso ra , dand o com o resu ltad o una p ro te ina m u tan te que tien e un resto de am in o a c id o m en os en co m p a ra c io n con la p ro te ina p recu rso ra . El te rm in o ta m b ie n pued e u tiliza rse en el co n te x to de una s e c u e n c ia de nu c le o tid o s , lo que s ig n ifica una d e le c io n o e lim in a c io n de un res to de la se cu e n c ia de p o lin u c le o tid o s de un p o lin u c le o tid o precurso r.

The rm in o "de le c io n", when it is used in the context of a se- lence of love, means one of the le o o le lim in ac About a remainder of the secu in the area of am in or of the rights of a person, giving or commissorship or a person so much so that he has a In the rest of the year, in com mpa ra c io n with p ro te ina p rec rso ra. The rm in o ta mb ie can be used in the context of a sequencing of nukes, which means one of the most important limitations of the a remainder of the sequence of some kind of a source or precursor.

Las e xp res ione s "p ro ce d e n te de" y "o b te n id o (a ) de" se re fie ren , en el co n te x to de una t io e s te ra s a p recu rso ra , a una t io e s te ra s a p ro du c ida por, o que puede p roduc ir, una cepa de l o rg a n ism o en cuestion , y ta m b ie n una t io e s te ra sa co d ifica d a po r una se cu e n c ia de p o lin u c le o tid o s a is lad a de d icha cepa y p ro d u c id a en un o rg a n ism o h o sp e d a d o r que co n tie n e d icha s e cu e n c ia de p o lin u c le o tid o s . A d ic io n a lm e n te , las e xp re s io n e s se re fie ren a una t io e s te ra s a que esta co d ifica d a po r una secu en c ia de p o lin u c le o tid o s de origen s in te tico y /o A D N c y que tien e las ca ra c te ris tica s de id e n tifica c io n de la t io e s te ra sa en cue s tion . P ara e jem p lifica r, " tio e s te ra sa s p ro ced en te s de la fam ilia E n te ro b a c te r ia c a e a (e n te ro b a c te ria s )" se re fie re a aq ue lla s e n z im a s que tie n e n ac tiv ida d tio e s te ra s a y que las e n te ro b a c te ria s p roducen de m a n e ra na tu ra l, a s i com o a tio e s te ra s a s com o las p ro du c ida s po r fu e n te s de e n te ro b a c te ria s pero que, u tilizan do te cn ica s de in g e n ie ria gene tica , se p ro du cen m ed ia n te o rg a n ism o s que no son e n te ro b a c te ria s tra n s fo rm a d o s con un po lin u c le o tid o que cod ifica d icha tioe s te rasa .

The "p ro ce den te de" and "ob te n id o (a) of" are recognized, in the context of an uncle is the case, To a guy he is sap ro du c ida by, or who can p roduc go, a strain of r r an an ism or in question, and also a guy is a co c tive of po r One of the leaders of the strain is that of the strain and who has been in an old age who is now aware of the importance of the strain. you. On a daily basis, the e xp re s ions refer to an uncle that is coded by a section of the population Ethical origin and / or DNA c and which have the most critical characteristics of the type in which the issue is in question. P ara e jem p lifica r, "uncle is te sp sa sp ro ced in te s of the family E n te ro bac te r ia caea (in te ro bac te ria s)" The senz im as they have ac tiv ida d unio es te ra say that the ones in ro bac te ria sp roducen in a way na tu ra l, so com un un un es es ra sas com o las p ro du c That is why you are going to be at the same time, but that, when you use genetic technology, you will be able to use it for a few years. They are not in the bac te ria s tra ns fo rm ados with a few lin uc le o tid o codi d dhaha tioe s rasa.

En el p re sen te d o cum e n to , las e x p re s io n e s "co n s tru cc io n de A D N " y "A D N tra n s fo rm a n te ", se u tilizan in d is tin ta m e n te pa ra re fe rirse a un A D N que se u tiliza para in tro d u c ir se cu e n c ia s en una ce lu la ho spe d a d o ra o en un o rg an ism o h o spe da do r. N o rm a lm e n te , una con s tru cc io n de A D N se ge ne ra m ed ia n te P C R in v itro o m ed ia n te otra te c n ic a o te c n ic a s a d ecu ad as con oc idas po r los exp e rtos en la m a te ria . En d e te rm in a d o s aspec tos , la con s tru cc io n de A D N co m p re n d e una s e cu e n c ia de in te res (p o r e jem p lo , una secu en c ia en tran te ). En a lgunas rea liza c io nes , la secu en c ia es ta un ida o p e ra tiva m e n te a e le m e n to s ad ic io na le s , ta le s com o e le m e n to s de con tro l (p o r e jem p lo , p ro m o to re s , etc.). U na con s tru cc io n de A D N puede co m p re n d e r a d e m a s un m a rc a d o r de se lecc ion . T a m b ie n puede co m p re n d e r una secu en c ia e n tra n te fla n q u e a d a po r ca jas de hom o log ia . En un a sp e c to a d ic io na l, la co n s tru cc io n de A D N c o m p re n d e o tras se cu e n c ia s no ho m o loga s , a n a d id a s a los ex tre m o s (p o r e jem p lo , s e cu e n c ia s re lle n a d o ra s o fla n q u e a n te s ). En a lg u n o s aspec tos , los e x tre m o s de la se cu e n c ia e n tra n te se c ie rran de ta l m an e ra que la co n s tru cc io n de A D N fo rm a un c ircu lo ce rrado . Las se cu e n c ia s tra n s fo rm a d o ra s pueden s e r de tip o s ilves tre , m u tan te o pueden es ta r m od ifica das . En a lg u n o s asp ec tos , la con s tru cc io n de A D N com p ren de se cu e n c ia s h o m o loga s a las del c rom o som a de la ce lu la ho spe da do ra . En o tros aspec tos , la con s tru cc io n de A D N co m p re n d e s e cu e n c ia s no hom o logas . Una ve z que la con s tru cc io n de A D N esta en sa m b la d a in v itro , esta puede u tiliza rse para: 1) in se rta r secu en c ias h e te ro lo g a s en una se cu e n c ia d ian a d e se a d a de una ce lu la ho spe da do ra ; 2 ) m u ta g e n iza r una reg ion de l c ro m o so m a de la ce lu la h o sp e d a d o ra (es decir, re e m p la z a r una secu en c ia e n d o ge na con una se cu e n c ia h e te ro loga ); 3) d e le c io n a r ge ne s d iana ; y /o (4) in tro d u c ir un p lasm id o de re p lica c io n en el hospedador.

In the case of cum in to, the "DNA-based DNA" and "DNA-related DNA" exp, they are used in different ways for men. Refer to a DNA that is used to introduce yourself in a given city or in a particular way of life. N o rm a lm in te, one with DNA DNA is generated in v PCR with another tenet tea or tea cn ic asad ecuad as with ears For the reasons in the matter. In terms of specific aspects, the DNA structure with a sequence of interest (for example, a section in transit). In some realizations, the sect in t is a one-way operative one to say to him, as well as to the others jem p lo, p ro mo to re s, etc.). A person with DNA DNA can understand a master of the lesson. You can also choose a section in which you are looking for homosexuality. In an ad hoc context, the understanding of DNA DNA afterwards is not only now, but also adds to the former (for example). I know that it is filled up only if they are). In some respects, the extremes of the sequence in question also make it clear that the construction of DNA fo rm to a closed circle . The signals that can be carried out can be of an important, many or many types of reasons. In some cases, the DNA con- struction with the responsibility of shom or logon is those of the city or the city of the city. In other respects, the DNA con - tribution of non-homologous DNA. Once the DNA construction is in place in the day, it can be used to: 1) in se se se se se r se c ias and the gas in a se c ia d ian ade se ada of a ce lu la ho spe da do ra; 2) a region of lc ro mo so ma de la ce lu la ho sp edado r (ie re re la zar a secu in the end o ge na with a se c a c a c on the he ro loga); 3) of le c io nar ge ne sd iana; and / or (4) in tro duc go a p pasm id o p plica c io n in the host.

S e d ice que un po lin u c le o tid o "co d ifica " un A R N o un po lip e p tid o si, en su es ta do na tivo o cua nd o se m an ip u la m ed ia n te m e tod os c o n o c id o s po r los e xp e rto s en la te cn ica , se puede tra n s c r ib ir y /o tra d u c ir para p ro d u c ir el A R N , el po lip ep tido , o un fra g m e n to de los m ism os. T a m b ie n se d ice que la cad en a a n tise n tid o de d icho po lin uc leo tido co d ifica las se c u e n c ia s de A R N o de l po lip ep tido . C o m o se sabe en la tecn ica , un A D N pu ede tra n s c r ib irs e m ed ia n te una A R N po lim e rasa para p ro d u c ir un A R N , y un A R N puede tra n s c r ib irs e m ed ia n te tra n sc rip c io n in ve rsa para p ro d u c ir un A D N . P o r tan to , un A D N puede c o d ifica r un A R N , y v ice ve rsa .

I know that a positive source or "codification" of an RNA or a potential lipid is, if it is in its state or when it is measured I know you know why you are in the technical field, you can work to go and / or work to go for the RNA, the lipid, or a Fragrance of the same. It is also noted that the body in which it has been published is the code of the terms of A R N or of the lipid lipid. As is known in the art, a DNA that can be transcribed into an RNA can be limited to an RNA, and an RNA can be transcribed into the environment. n sc rip c io n in ve rsa for p ro duc go a DNA. P o r so, an A D N can c or d an A R N, and see ice.

La fra se "e q u iva le n te " en este con tex to , se re fie re a e n z im a s t io e s te ra sa s que estan co d ifica d a s po r un p o lin u c le o tid o ca p a z de h ib rid a rse con el p o lin uc leo tido que t ie n e la se cu e n c ia de la S E C ID NO: 31 exp ue s ta en la FIG. 58, en co n d ic io n e s de m ed io a m a x im a rig u ro s id a d . P o r e jem p lo , s e r eq u iva le n te s ig n ifica que una tio e s te ra sa m ad u ra e q u iva le n te co m p re n d e al m e n o s 70 %, al m en os 75 %, al m en os 80 %, al m en os 85 %, al m e n o s 90 %, al m en os 91 %, al m en os 92 %, al m en os 93 %, al m en os 94 %, al m enos 95 %, al m en os 96 %, al m en os 97 %, al m en os 98 % y /o al m en os 99 % de id en tidad de se cu e n c ia con la se cu e n c ia de a m in o a c id o s de la S E C ID NO: 31 e xp u e s ta en la FIG. 58.

The phrase "equ iva le n te" in this text, it is true that it is known that they are coded for a little bit of peace or peace. rid with the po l u u tide that has the meaning of SEQ ID NO: 31 set forth in FIG. 58, in the midst of m o r a m a x im a rig u ro s id a d. P o jem p lo, being effective means that an uncle is more equitable at least 70%, at least 75%, at m in os 80%, to m in os 85%, at least 90%, to m in os 91%, to m in os 92%, to m in os 93%, to m in os 94%, at least 95 %, at m in 96%, at m in os 97%, at m in os 98% and / or at m in os 99% of the identity of the person in charge with the one in am am o The codes of SEQ ID NO: 31 are shown in FIG. 58.

U na "e s te r s in ta sa " es un pep tido cap az de c a ta liz a r una reacc ion b ioq u im ica para p ro d u c ir es te res. P o r e jem p lo , una e s te r s in ta sa es un pe p tido que puede p a rtic ip a r en la tra n s fo rm a c io n de un t io e s te r a un e s te r graso . En d e te rm in a d o s aspec tos , un e s te r s in ta sa tra n s fo rm a en tioe s te r, una ac il-C oA , en un e s te r graso . En un asp ec to a lte rna tivo , una e s te r s in ta sa u tiliza un t io e s te r y un a lcoh o l com o sus tra to s para p ro d u c ir un e s te r graso . Las es te r s in ta sa s son cap ace s de u tiliza r a c il-C o A de cad en a corta y la rga com o sustra to . A d e m a s , las e s te r s in ta sa son c a p a ce s de u tiliz a r a lco h o le s de ca d e n a co rta y la rga co m o sus tra to . C o m o e je m p lo s no lim ita n te s de e s te r s in ta sa s se in c luye n ce ra s in tasas , e s te r ce ra s in ta sa s , acy l-C o A : a lco h o l tra n sa c ila sa s , a c iltra n s fe ra sa s , ac il co e n z im a grasa : a lco h o l a c iltra n s fe ra sa s grasas, a c il-A C P tra n s a c ila s a g rasa y a lcoh o l ace tiltra n s fe ra sa . U na e s te r s in ta sa que tra n s fo rm a un t io e s te r de a c il-C o A en una ce ra se d e n o m in a ce ra s in tasa . Las e s te r s in ta sa s e je m p la re s in c luyen las c la s ifica d a s con el nu m ero de c la s ifica c io n de en z im a s EC 2.3.1.75. la exp res ion "e s te r s in ta sa " no com p ren de e n z im a s que ta m b ie n tie n e n ac tiv id a d tio e s te ra sa . En el p re se n te do cu m e n to , las e n z im a s que tie n e n ac tiv id a d ta n to e s te r s in ta sa com o tio e s te ra sa , se ca te g o riza n com o tioe s te rasa s .

A "e s te rs in ta sa" is a peptic capital of a kind of a biochemical or reaction for this purpose. P o je je p p, an e s te rs in ta sa is a peptide that can be seen in the path of an uncle that is a fatty one. In terms of specific aspects, a person will be involved in uncle, an ac- c oA, in a fatty environment. In an aspiring way, one of you will use an uncle and a person who will try to get a fatty oil. These are the capitals of the use of rac il-C or A in short and the common law. In addition, these schools are capable of using the help of Cana Corta and the Rga as their treatment. How do you know what is wrong with you? Do you want to pay in taxes, do you want to know, here? lco hol tra n sa c ila sa s, ac iltra ns fe ra sa s, ac il co enzy im a gre: to lco holac iltra ns fe ra sa s fats, ac il-A CP tra nsac ila sag rasa and lcoh ol ace tiltra ns fe ra sa. U na e s te rs in ta sa that trans a un tio e s te r ac ac-C or A in a ce is called a ce ra s in rate. The te s rs in ta sa is the re s in c ls the c u ss with the number of c the meaning of in EC 2.3.1.75. The term "e s te rs in ta sa" does not include enzymes that also have an ac- tiv id ad uncle es te ra sa. In the past, the enzymes that have ac tiv id ad ta n to e s te rs in ta sa com o uncle es te ra sa, se ca r la riza n com o tioe s rasa s.

La exp res ion "g en es e xp re sa d o s " se re fie re a ge ne s que se tra n sc r ib e n en A R N m e n sa je ro (A R N m ) y que de spues se tra d u c e n en p ro te inas , as i co m o a genes que se tra n sc rib e n en tip o s de A R N , ta le s com o A R N de tra n s fe re n c ia (A R N t), A R N r ib o so m ico (A R N r) y A R N regu lado r, que no se tra d u ce n en p ro te inas.

The expression "g en es e xp re sa dos" refers to people who are sc r ib en in ARN men sa je ro (mRNA) and then translated into p ro The genes, as well as genes that are treated in RNA types, such as RNA of trans fe fe n re rn (RNA t), RNA r ib or only (RNA r) and RNA regu side r, which is not translated in p ines.

Las exp re s io n e s "ca se te de e xp res ion " o "v e c to r de exp res ion ", se re fie ren a una con s tru cc io n de po lin u c le o tid o s ge ne ra d a de fo rm a reco m b in an te o s in te tica , con una serie de e le m e n to s e sp e c ifico s que pe rm iten la tra n sc rip c io n de un po lin u c le o tid o p a rticu la r en una ce lu la d iana. Un case te de exp res ion reco m b in an te puede in co rp o ra rse en un p lasm ido , en un c rom o som a , en un A D N m itoco nd ria l, en un A D N p lasm id ico , en un v iru s o en un fra g m e n to de po lin u c le o tid o . N o rm a lm e n te , la pa rte de case te de e xp re s io n re co m b in a n te de un v e c to r de e xp re s io n inc luye , e n tre o tras se cu e n c ia s , una secu en c ia de p o lin u c le o tid o s que se va a tra n s c r ib ir y un p rom oto r. En asp ec tos pa rticu la res, los v e c to re s de e xp re s io n tie n e n la ca p a c id a d de in c o rp o ra r y e x p re s a r fra g m e n to s de p o lin u c le o tid o s h e te ro lo g o s en una ce lu la ho spe da do ra . En el co m e rc io se d ispo ne de m uch os ve c to re s de exp res ion p ro ca rio tas y eu ca rio tas . La se lecc io n de ve c to re s de exp res ion ap ro p ia d o s esta de n tro de l co n o c im ie n to de los exp e rtos en la te cn ica . En el p re sen te d o cum e n to , las e x p re s io n e s "ca se te de exp res ion ", "co n s tru cc io n de A D N " y sus e q u iva le n te s g ra m a tica les , se u tilizan in d is tin tam en te .

The expi sions "ca se te de e xp res ion" or "neighbor to exp exp ion", refer to one of the structures of potential control It has been for the first time in technical terms, with a series of reports that allow the tram sc rip tio n of a po lin uc le or tid opa rticu the r in a ce lu the day. A case of reco mb in an te res expression can be included in a plasmid, in a rom or som, in a medical DNA, in a medical DNA, in a future, in a fragance de po lin uc le o tid o. N o rm a lm en te, the case of e xp re s io n re co mb in an te of a neighbor of e xp re s io n inc luye, between three or three years Yes, a section of the population that is going to be carried out and a period of autumn. In this country, the residents of the country have the right to be in charge of the fraternity of the French community. Ro lo gos in a ce lu la ho spe da do ra. During the period, there are many times of expiration for public and eu. The lesson of the right-of-way view is within the context of the reports in the technical field. In the case of cum in to, the exp "s" will be exp eration "," DNA con tr e ction "and their mathematical equation, they are used in d is tin tam in te.

La exp res ion "d e riva d o de ac ido graso", ta l com o se usa en el p resen te d o cum e n to , se re fie re a una com p os ic ion que p rocede de una ru ta m e tab o lica , cuya ruta in c luye una reacc ion de tio e s te ra sa . P o r tan to , los p roduc tos p ro ce d e n te s de ac ido s g ra sos pueden s e r p ro du c to s que son, o p roceden de, ac ido s g ra sos o es te res g rasos, que son p ro d u c to s de una rea cc io n de tio e s te ra sa . P o r tan to , co m o d e riva d o s de ac ido s g ra sos se inc luyen , po r e jem p lo , p ro du c to s que son, o que p roceden de, ac ido s g ra sos que son el p ro du c to de reacc ion d irec ta de una tioe s te rasa , y /o de un e s te r g raso que es un p ro du c to de rea cc io n d irec ta de una tio e s te ra sa . C om o de riva dos de ac idos g rasos e je m p la re s se in c luyen , po r e jem p lo , a lco h o le s de ca d e n a co rta y la rga , h id ro ca rb u ro s , y a lco h o le s g ra sos y es te res, in c lu ye n d o ceras, es te res de ac ido s g ra sos y/o es te res g rasos. C o m o e jem p los no lim ita tivo s e sp e c ifico s de d e riva d o s de ac ido s g ra sos se inc luyen , ac ido s g rasos, e s te re s m e tilico s de ac ido s g rasos, e s te re s e tilico s de ac ido s g rasos , a lcoh o le s g rasos, a lq u il-a ce ta to s g rasos , a ld e h id o s g rasos , a m in a s g rasas , am ida s g rasas, su lfa tos grasos, e te re s g rasos, ce tonas , a lcanos , o le fin a s in te rnas, o le fin as te rm in a le s , ac ido s d ica rb o x ilico s , ac ido s u>-d icarboxilicos, d io le s y ac ido s g ra sos te rm in a le s y /o in te rnos.

The expression "of the fatty acid source", as it is used in the past, refers to a com- pany that follows a route. lica, whose route includes a reaction from uncle is te ra sa. Therefore, the products for which they can continue to be that they are, or that they are, for that reason or that are certain, which are Ro duc to s of an uncle's reaction is te ra sa. P o so to, co mode riva dos de ac gone sg ra sos are included, p re jem p lo, p ro du c to s who are, or p roce of, ac cted sg ra are those who are the p ro The direct reaction of an uncle is clear to you, and / or of a person who is a direct reaction of an uncle. C om o de riva dos de acid aids e je mp la re s in c luyen, po re jem p lo, a lco ho le s de cana dena co rta and la rga, h id ro ca rb u ro s , and now you are and that you are, in waxing waxes, this is what you are saying and / or this is your reason. How do you think that you are the only one of the two countries of the country? They are included, they are still ordinary, they are typical of the city, they are It is typical of the following, to lcoh or the following, to lq u il-a ta ta sat, to ld eh id osg satos, am in asg rasas, am ida sg rasas, his lfa fatty coughs, he re you sgs, ce tonas, lcanos, or the end as in te rnas, or the end so rm in a s s, c i d d r b,,,,,, a a a, su le sy ac ido sg ra sos te rm in a le sy / o in te rnos.

La exp res ion "e n z im a s de riva d a s de ac ido s g ra sos" se re fie re a, con ju n ta e in d iv idu a lm e n te , en z im a s que pueden exp re sa rse o so b re e xp re sa rse en la p ro du cc ion de de riva d o s de ac ido s grasos. E stas en z im a s pueden fo rm a r parte de una ru ta b ios in te tica de ac ido s grasos. C o m o e jem p los no lim itan te s de s in ta sas de riva das de ac ido s g ra sos se in c luye n s in ta sas de ac ido s g rasos, t io e s te ra sa s , a c il-C o A s in tasas , a c il-C o A red uc tasa s , a lcoh o l d e sh id rog en asa s , a lcoh o l ac iltran s fe rasas , a c il-C o A red uc tasa fo rm a d o ra de a lcoh o l graso , d e sca rb o x ila sa s de ac ido s grasos, ac ido ca rb o x ilico reduc tasas , ace til tra n s fe ra s a s de a lcoh o l g raso y e s te r s in tasas. Las e n z im a s de riva das de ac idos g ra sos tra n s fo rm a n los sus tra to s en de riva d o s de ac ido s grasos. En d e te rm in a d a s c ircu ns tan c ias , un sus tra to a d e cu a d o puede s e r un p r im e r de riva do de ac ido graso , que se tra n s fo rm a m ed ia n te una en z im a de riva da de ac ido graso , en un seg u n d o de riva d o de ac ido g raso d ife ren te .

The expression "enzymes of riva das de ac ido sg ra sos" is true, as long as it is possible, in areas that may expire. B re e xp re sa rs p o du r ion de de riva dos de ac gone s fatty. These are likely to be part of a fatty route of fatty acids. How do you know what is happening in Sivas de Riva das de Acido Sg Ra SOS is in C Luye Ns in Ta Sas de Acido Sg Rasos, Uncle is Te Ra Sa S, Ac il-C o A s in rates, ac il-C or A red uc rate s, to lcoh olde sh id rog in asa s, to lcoh ol ac iltran s fe rasas, ac il-C or A red uc rate for rm adora of A lcoh ol graso, from sca rb ox ila sa s de ac gone s oily, ac c a ca rb ox illico reduced rates, acetil tra ns fe ra sas de a lcoh olg sato and e s rs in rates. The g of acid acids are the main sources of their fatty acid treatments. In te rm in adasc ircuns so c ias, a proper treatment can be a first of fatty acid, which is for the most part of the year. Riva da of fatty acid, in a sec- ond of riva do of greasy acid.

La e xp res ion "a lco h o l g ra so " se re fie re a un a lcoh o l que tie n e la fo rm u la R O H . En d e te rm in a d o s asp ec tos , un a lcoh o l g ra so es un a lcoh o l fa b rica d o a p a rtir de un ac ido g raso o de riva do de ac ido graso . En un aspecto , el g rupo R tien e una lo ng itud de al m en os a p ro x im a d a m e n te 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 o 20 ca rbonos . El g ru p o R puede te n e r una cad en a linea l o ram ificada . Las cad en as ra m ifica d a s pueden te n e r uno o m as pun tos de ram ificac io n . A de m as , las ca d e n a s ra m ifica d a s pueden in c lu ir ram as c ic licas, ta le s com o g rupos de c ic lo p ro p a n o o epox ido . A s im ism o , el g rupo R puede e s ta r sa tu ra do o insa tu rado . Si esta insa tu rado , el g rupo R puede te n e r uno o m as pu n tos de insa tu ra c ion . En una rea liza c io n , el a lcoh o l g ra so se p ro du ce b io s in te tica m e n te . Los a lco h o le s g ra sos tie n e n m u ch o s usos. P o r e jem p lo , los a lcoh o le s g ra sos pueden u tiliza rse para p ro d u c ir p ro du c to s qu im icos esp ec ia le s . E spe c ificam en te , los a lcoh o le s g ra sos pueden u tiliza rse com o b ioco m bus tib le s ; com o d iso lve n te s para g rasas, ceras, go m as y res inas; en ba lsa m os fa rm a ce u tico s , e m o lie n te s y loc iones; com o ad itivos para ace ites lu b rica n tes ; en de te rg e n te s y em u ls io n a n te s ; com o ag e n te s te x tile s a n tie s ta tico s y de acabado ; com o p las tifican tes ; com o te n s io a c tiv o s no ion icos; y en cosm e tica , po r e jem p lo , com o esp esa n tes .

The expression "a lco h o l g ra so" refers to a lcoh or l who has the form rm u R O H. In d e te rm in a d o s asp ec tos, an a lcoh or l g ra so is an a lcoh o l fa b rica d o a p a part of a private or a riva do of fatty acid. In one aspect, the group R has the best of all in 5 months, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 , 18, 19 or 20 ca rbonos. The group may have a cad in a branched line. The certified bodies can have one or more branching points. On the other hand, the ca m e m a s ra m ifica d a s may include cyclic ramifications, as well as the rupture of the capital or epoxy. In addition, the R group can be your own or insane. If your degree is insane, the R group may have one or more points to insure your ratio. In a realization, the a lcoh or l g ra so p ro du ce b io s in tica m e n te. There are many uses for many years. P o r e jem p lo, the a lcoh or le s g ra sos can be used for the special chemical sp e r. In fact, the people who can be used can be used as a bus or bus; How to isolate vents for scratches, waxes, gums, and res; in fact, it is more important, and it is loc ions; com o adives for ace acees lu b rica n tes; in de te rg e n te s and em u ls io n a n te s; com o ag e n te s te x tile s a n tie s tatico s s y finishing; com or p tify them; as a non-ionic sector; and in cosmetics, po r e jem p lo, com o esp that n tes.

La e xp res ion "p e p tid o s fo rm a d o re s de a lcoh o l g ra so " se re fie re a pe p tid os c a p a ce s de c a ta liz a r la co n ve rs io n de ac il-C o A en a lcoh o l graso , in c luye nd o la a c il-C o A red uc tasa fo rm a d o ra de a lcoh o l g ra so (FA R , EC 1.1.1. *), la ac il-C o A re d u c ta sa (E C 1.2.1.50 ) o la a lco h o l d e s h id ro g e n a s a (E C 1.1.1.1 ). A d ic io n a lm e n te , un e xp e rto ha b itu a l en la m a te ria a p rec ia ra que a lgu nos pep tidos fo rm a d o re s de a lcoh o l g ra so ca ta liza ran ta m b ie n o tras reacc iones . P o r e jem plo , a lg u n o s pe p tid os de a c il-C o A red u c ta sa a ce p ta ra n su s tra to s d is tin tos de ac idos g rasos. P o r lo tan to , ta m b ie n se in c luye n d icho s pep tidos no esp ec ificos . En la te cn ica se con oce n s e cu e n c ia s de po lin u c le o tid o s que cod ifican p e p tid o s fo rm a d o re s de a lcoh o l graso , y d icho s pe p tid os estan d isp o n ib le s al pub lico .

The "p e p tid os fo rm ado re s de a lcoh olg ra so" section is now known as a layer of ca l ca -C or A in a lcoh ol fatty, in c luye nd or the ac il-C or A red uc fo rm adora rate of a lcoh olg ra so (FA R, EC 1.1.1. *), The ac il -C or A re duc ta sa (EC 1.2.1.50) or the a lco holdesh id ro genasa (EC 1.1.1.1). In this day, an e xp e rto ha b itu al in the ma ap ria ap rec ia ra that some of the people who have been interested in lcoh olg ra so ca ta liza ran ta mb I don't go after reactions. P o je je plo, a few periods of ac c-C o A red uc ta sa to ce p ta ra ns its treatment of acidic acids. For the most part, there is also a lack of evidence. In the technical field, it is known that there are some coding codes for fatty acids, and that they are only available. on ib s s to the public.

La exp res ion "a ld e h id o g ra so " se re fie re a un a ld e h id o que tie n e la fo rm u la R C H O c a ra c te riza d a po r un grupo ca rb o n ilo in sa tu ra do (C =O ). En d e te rm in a d o s aspec tos , un a ld e h id o g raso es un a ld e h id o fa b rica d o a pa rtir de un a c id o g ra so o d e riva d o de a c id o g ra so . En un asp ec to , el g ru p o R t ie n e una lo ng itud de al m e n o s a p ro x im a d a m e n te 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 o 20 ca rb on os . El g ru p o R pued e te n e r una ca d e n a linea l o ram ificada . Las ca d e n a s ra m ifica d a s pueden te n e r uno o m as pu n tos de ram ificac io n . A d e m a s , las cadenas ra m ifica d a s pueden s e r ra m ifica c io n e s c ic licas. A s im ism o , el g ru p o R puede e s ta r sa tu ra d o o in sa tu ra do . Si esta in sa tu ra do , el g rupo R puede te n e r uno o m as pun tos de insa tu ra c ion . En un aspecto , el a lde h ido g raso se p roduce b io s in te tica m e n te . Los a ld e h id o s g ra sos tie n e n m u ch o s usos. P o r e jem p lo , los a ld e h id o s g ra sos pueden u tiliza rse pa ra p ro d u c ir p ro d u c to s q u im ico s esp e c ia le s . E sp e c ifica m e n te , los a ld e h id o s g ra so s pu ed en u tiliza rse pa ra p ro d u c ir po lim e ro s , res inas, tin te s , a ro m a tiza n te s , p las tifica n tes , pe rfum es, p ro du c to s fa rm a ce u tico s y o tros p roduc tos qu im icos . A lg u n o s se u tilizan com o d iso lve n tes , co n se rva n te s o de s in fe c ta n tes . A lg u n o s co m p u e s to s na tu ra les y s in te ticos , ta le s com o v ita m in a s y ho rm onas, ta m b ie n son a ldeh idos .

The expression "to ld eh id og ra so" refers to a ld eh id or who has the form of the right to give a group of people in sa tu tu ra do (C = O). In d e te rm in a d o s aspec tos, a year is a factory that is a country that is part of a specific area of interest. In an asp e ct, the group R t ie ne a ng itud of at least 1 x 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 ca rb on os. The group may have a branched line. The ca m e m a s ra m ica d a s may have one or more branches of ramification. On the other hand, the branded chains can only be cyclic. A s im ism o, g ru p o R can be s sa r sa o ra ra o o in sa tu ra do. If it is in sa ra do, the group R may have one or more points of insa t ration. In one aspect, the senior citizen is produced in the most technical way. The years have been used. P o r e jem p lo, a ld e h id o s g ra sos can be used for the same time as only. E sp e c tive men, the a ld eh id osg ra so s pu ed en u tiliza rse para ra p ro duc go po lim e e s, res inas, tin te s, a ro ma tiza n te s, p The qualifiers, for example, are more important and only chemical products. Some of them are used as isolates, as they are known in faiths. A co m p u e s to s na tu ra les and s in te ticos, ta s com o v ita m in a s and ho rm onas, ta m b ie are a ldeh idos.

En el p re sen te do cu m e n to , las e x p re s io n e s "p o lip e p tid o b io s in te tico de a ld e h id o g ra so " "a c id o c a rb o x ilico reduc tasa " y "C A R ", se u tilizan in d is tin tam en te .

In the past, the expi sions "po lip e p tid ob io s in tetico of a ld eh id og ra so" "a c id oca rb ox ilico reduced rate" and "CAR" is used in d is tin tam in te.

La exp res ion "e s te r g raso" se re fie re a un e s te r que t ie n e m as de 5 a to m o s de carbono . En d e te rm in a d o s aspectos, un e s te r g ra so es un e s te r fa b rica d o a p a rtir de un ac ido graso , po r e jem p lo , un e s te r de ac ido graso. En una re a liza c io n , un e s te r g ra so con tiene un lado A (es dec ir, la ca d e n a de c a rb o n o un id a al o x ig e n o de ca rb o x ila to ) y un lado B (es decir, la cad en a de ca rb o n o que co m p re n d e el c a rb o x ila to p recu rso r). En un a sp ec to pa rticu la r, cua nd o un e s te r g ra so p ro ce d e de la ru ta b io s in te tica de a c id o s g rasos , el lado A ap o rta un a lcoho l, y el lado B a p o rta un ac ido g raso . S e pu ed e u tiliz a r c u a lq u ie r a lco h o l pa ra fo rm a r el lado A de los es te re s g rasos . P o r e jem p lo , el a lco h o l puede p ro c e d e r de la ru ta b io s in te tica de los ac ido s g rasos. C o m o a lte rna tiva , el a lcoh o l puede p ro du c irse a tra v e s de ru tas b io s in te tica s de ac idos no grasos. A de m as , el a lcoh o l puede p ro p o rc io n a rse de m an era exogena . P o r e jem p lo , el a lco h o l puede su m in is tra rse al ca ldo de fe rm e n ta c io n en los caso s en los que el e s te r g ra so lo p roduce un o rg an ism o . C om o a lte rna tiva , un ac ido ca rb ox ilico , ta l com o un a c id o g ra so o a c id o ace tico , puede s u m in is tra rse de m an e ra exo ge na en los caso s en los que el e s te r g ra so lo p roduce un o rg a n ism o que ta m b ie n puede p ro d u c ir a lcoho l. Las cad en as de ca rb on o que com p ren den el lado A o el lado B pueden te n e r cu a lq u ie r long itud . En un asp ec to , e l lado A de l e s te r t ie n e una lo ng itud de al m en os a p ro x im a d a m e n te 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 16, 18 o 20 ca rb on os . El la do B de l e s te r tien e una lo ng itud de al m en os a p ro x im a d a m e n te 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24 o 26 ca rbonos . El lado A y/o el lado B pueden s e r de cad en a linea l o ram ificada . Las ca d e n a s ram ificada s pueden te n e r uno o m as pun tos de ram ificac io n . A de m as , las cad en as ram ificada s pueden in c lu ir ram ificac io ne s c ic licas , ta le s com o g ru po s de c ic lo p ro p a n o o ep ox ido . A d e m a s , el lado A y /o el lado B pueden e s ta r sa tu ra d o s o in sa tu ra dos . Si es ta in sa tu ra do , el lado A y /o el lado B puede te n e r uno o m as pu n tos de insa tu ra c ion . En una rea liza c io n , el e s te r g raso se p roduce b io s in te tica m e n te . En este aspecto , p rim ero se "a c tiva " el ac ido graso. E je m p lo s no lim ita n te s de ac ido s g ra so s ac tiva d o s son ac il-C oA , ac il A C P , a c il-A M P y ac il fos fa to . La a c il-C o A puede s e r un p ro d u c to d irec to de la b io s in te s is o d e g ra d a c io n de los ac ido s g rasos. A de m as , la a c il-C o A pued e s in te tiza rse a p a rtir de un ac ido g ra so libre, una C o A y un tr ifo s fa to de nu c le o tid o de a d e n o s in a (A T P ). Un e jem p lo de una e n z im a que p roduce a c il-C o A es una a c il-C o A s in tasa . Una ve z que se ac tiva el ac ido graso , se puede tra n s fe r ir fa c ilm e n te a un nuc le o filo recep to r. Los nu c le o filo s e je m p la re s son a lcoh o le s , tio les , am ina s o fos fa tos . En o tro aspec to , el e s te r g raso puede p ro ce d e r de un a c ilt io e s te r g raso y un a lcoho l. En un aspecto , el e s te r g raso es una cera . La ce ra puede p ro ce d e r de un a lcoh o l g raso de cadena la rga y de un ac ido g ra so de cad en a la rga . En otra rea liza c io n , el e s te r g raso es un t io e s te r de ac ido graso , po r e jem p lo , ac il C o e n z im a A (a c il-C o A ) grasa . En o tros asp ec tos , el e s te r g ra so es un ac il p a n to te n a to g raso , una ac il p ro te ina tra n s p o rta d o ra de ac ilo (ac il-A C P ), un es te r de e n z im a ac ilo g ra so o un e s te r de fo s fa to g raso . Un e s te r puede fo rm a rs e a p a rtir de un p ro du c to in te rm e d io de e s te r de e n z im a acilo a tra ve s de la a lcoh o lis is del en la ce e s te r para fo rm a r un nuevo e s te r y la e n z im a libre. Los e s te re s g ra sos tien en m uch os usos. P o r e jem p lo , los es te res g ra sos pueden u tiliza rse com o un b io co m b u s tib le o com o un te n s io a c tivo , o com o un co m p o n e n te de estos.

The term "e s te r g sato" refers to an e s te r that has more than 5 to m or s of carbon. In these aspects, an example is a factor that is rich in a fatty acid, because it is a fatty acid. In a review, one of you has a side A (that is, the channel of the channel is an ID of the oxidized channel) and a side B (that is, the cad in a ca rb ono that com the p r rn ox o ila to p rec rso r). In one year of the year, when a person is responsible for the roadside routines, the side A ap o rta a year lcoho l, and the B side supports a large area. I can use r r u u l r u ie r a lco h o l pa ra fo rm a r side A of these re sgs. P o r e jem p lo, the a lco h o l can p ro c e d e r of the technical route of the sacred aids. C o m o a lte rna tiva, the a lcoh o l can p ro du c go through b ios s in ticas s ticas of non-fatty acids. In addition, the year may also be exogenous. P er jem p lo, the a lco hol can be measured in the case of faith rm ta ta io n in the cases in which he or she is only able to produce a rg an ism o. C om o lte rna tiva, a ca rb ox ilic acid, such as an ace or id a c act, can be added in a foreign manner in the In those cases in which he is only responsible for one or more years, he may also be able to go to Lima. Carcasses that are included in either side A or side B can be used for a long time. In an asp e, the A side of it is a matter of interest in the first 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 16, 18 or 20 ca rb on os. On the B of the latter you will have a lot of interest in the 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24 or 26 ca rbonos. Side A and / or Side B can be found in a straight line or branched line. Branches may have one or more branching points. In addition, branched bodies can be included in the cyclical, as well as in the cases of episodes. Moreover, side A and / or side B can be s ra r sa s d o s or in sa tu ra ra two. If this is your reason, Side A and / or Side B may have one or more points of use. In a realization, the state is produced in Mexico. In this regard, first the fatty acid is "active." E je m p lo s no lim ita n te s n ac s g ra so s ac tiva d o s are ac il-C oA, ac il A C P, a c il-A M P and ac il fos fa to. The a c il-C or A can be a p ro d u c a d irec of the b io s sin te s is o d e g ra d a c io n of the sacred aids. In addition, the ac-C or A can be used to move away from a free, a C or A, and a three-way traffic system or in addition to (ATP). An example of an e n z im a that produces a C-C or A is a C-C or A s in rate. Once the fatty acid is activated, it can be done easily to a new recipient. The nu s le je philo s e je m p la re s are a lcoh o le s, uncle les, am ina s or fos fa tos. In other respects, the latter can only be one year later and one year later. In one aspect, this is a wax. The wax can be from one year to another, or the chain satin, and from a large group of people in the country. In another realization, this is a type of fatty acid, because it is fatty, A / C or E n z im a A (a c il-C or A) fat. In other cases, this is a common plan, a joint venture will be a step by step (ac-A-CP), a This is an enzyme that is one or more important. An e s te r can be a part of a project from which it is also available from the acyl to the acyl through the a lcoh or lis es del in the ce e e s te r for fo rm ar a new e s te ry the free enzyme. This is what you have in many uses. P o r e jem p lo, these rents can be used as a co b bio b tibial or com o a te s s io s tio cive, or as a co m p o n e n te of these.

La exp res ion "e s te r g ra so fre n te a o tros d e riva d o s de ac ido s g rasos", com o se usa en el p resen te d o cum e n to , se re fie re al re n d im ie n to p ro p o rc io n a l de e s te r g ra so en co m p a ra c io n con la ca n tid a d to ta l de o tro s d e riva d o s de ac ido s g ra sos que no son es te res g rasos. En o tras pa lab ras, la can tidad de es te res g ra sos se com p ara con la can tidad de d e riva d o s de ac ido s g ra sos d is tin to s de e s te re s grasos.

The expression "e s te rg ra so fre n te ao tros de riva dos de acido sg rasos", as used in the past session, refers to the re nd im ie n To be able to support you in cooperation with the fallout of the second two years of life, these are not the reasons for this. . On or after the words, the amount of these resources is compared with the amount of money that is available to you.

Las e xp res ione s "p ro d u c tiv id a d de fe rm e n ta c io n " o "p ro d u c tiv id a d ", se re fie ren a la v e lo c id a d de p ro du cc ion del p ro du c to y se exp resa en g l-1h '1. La p ro du c tiv ida d e sp ec ifica es la p ro du c tiv ida d n o rm a liza d a para la c o n ce n tra c io n de c a ta liz a d o r y se exp re sa com o g/g l-1h '1 g (c a ta liz a d o r) '1.

The e xp res ione s "p ro duc tiv id ad de fe rm en ta c io n" or "p ro duc tiv id ad", are recognized as the id id of p r du duction of the p ro du c to and exp e in g l-1h '1. The p ro du c tiv ida of sp e cfica is the p ro du c tiv ida dno rm a liza da for the conca ntra tio n ca t liz adory is exp re sa com og / g l-1h ' 1 g (ca l liz ador) '1.

Las exp re s io n e s "titu lo de fe rm e n ta c io n " o "titu lo ", se re fie ren a la co n ce n tra c io n de un p ro du c to de reaccion, g e n e ra lm e n te e xp resa da com o g/l pero ta m b ie n en o tras un idades (es decir, m o la r, m asa /m a sa , m a sa /vo lu m e n o vo lu m e n /vo lu m e n ).

The terms "title of faith rm in ta tio n" or "title" refer to the connection to a reaction point, generally lm in te e xp resa da com og / l but ta mb ie n in or after units (i.e., r r, m asa / ma sa, ma sa / vo lu meno vo lu men / vo lu men).

La exp res ion "re n d im ie n to de fe rm e n ta c io n " o el te rm in o "re n d im ie n to ", se re fie ren a la can tida d de p roducto p ro d u c id o a p a rtir de una can tidad dada de m a te ria p rim a y g e n e ra lm e n te se exp resa co m o la re lac ion de m asa del p ro d u c to p ro d u c id o d iv id id o en tre la m asa de m a te ria p rim a co n su m id a (g de p ro d u c to /g de m a te ria prim a). T am b ien pued e e x p re s a r un ren d im ie n to m o la r (m o les p ro d u c to /m o le s de m a te ria prim a).

The expression "re nd im ie n to of faith rm en ta c io n" or the tea rm in or "re nd im ie n to", refers to the amount of p ro duc product. id opa r rtir of a given amount of ma ma ria p rim aygene ra lm in which it is explained as the re ion of the handle of the du du to p ro duc id od iv id id or between the handle ma p rim p rim a co n its m id a (g de p ro duc to / g de ma te ria prim a). It is also possible to express a change in the rhe r (mo p p du du to / mo le s de ma ma ria prim a).

La expresion "fraccion de carbono moderno" se refiere al parametro ' V segun lo definido por el Instituto Nacional de Estandares y Tecnologia (NIST) en Materiales de Referencia Estandar (SRM) 4990B y 4990C, conocido como estandares de acidos oxalico HOxI y HOxII, respectivamente. La definicion fundamental se refiere a 0,95 veces la relacion isotopica 14C/12C HOxI (referenciado a AD 1950). Esto es muy equivalente a la madera previa a la revolucion industrial con deterioro corregido. Para la biosfera viva actual (material vegetal), la fM es de aproximadamente 1,1.

The term "modern carbon fraction" refers to the 'V parameter as defined by the National Institute of Standards and Technology (NIST) in Standard Reference Materials (SRM) 4990B and 4990C, known as HOxI and HOxII oxalic acid standards, respectively. The fundamental definition refers to 0.95 times the 14C / 12C HOxI isotopic ratio (referenced to AD 1950). This is very equivalent to wood prior to the industrial revolution with corrected deterioration. For the current living biosphere (plant material), the FM is approximately 1.1.

La expresion "ensayo funcional" se refiere a un ensayo que proporciona una indicacion de la actividad de una proteina. En aspectos particularmente preferidos, la expresion se refiere a un sistema de ensayo en donde se analiza una proteina para determinar su capacidad natural para funcionar. Por ejemplo, en el caso de enzimas, un ensayo funcional implica determinar la eficacia de la enzima para catalizar una reaccion.

The term "functional assay" refers to an assay that provides an indication of the activity of a protein. In particularly preferred aspects, the expression refers to an assay system where a protein is analyzed to determine its natural ability to function. For example, in the case of enzymes, a functional assay involves determining the efficacy of the enzyme to catalyze a reaction.

"Gen" se refiere a un polinucleotido (por ejemplo, un segmento de ADN), que codifica un polipeptido y que incluye regiones que preceden y siguen a las regiones codificantes, asi como a secuencias intermedias (intrones) entre segmentos codificantes individuales (exones).

"Gen" refers to a polynucleotide (for example, a segment of DNA), which encodes a polypeptide and that includes regions that precede and follow the coding regions, as well as intermediate sequences (introns) between individual coding segments (exons) .

La expresion "genes homologos" se refiere a un par de genes de especies diferentes pero relacionadas, que se corresponden entre si y que son identicos o similares entre si. La expresion abarca genes que estan separados por el proceso de especiacion durante el desarrollo de nuevas especies (por ejemplo, genes ortologos), asi como genes que se han separado por duplicacion genetica (por ejemplo, genes paralogos).

The term "homologous genes" refers to a pair of genes from different but related species, which correspond to each other and are identical or similar to each other. The expression encompasses genes that are separated by the speciation process during the development of new species (for example, orthologous genes), as well as genes that have been separated by genetic duplication (for example, paralogic genes).

La expresion "proteina endogena" se refiere a una proteina que es nativa para una celula, o que se produce en ella de manera natural. Un "polinucleotido endogeno" se refiere a un polinucleotido que esta en la celula y no se introdujo en la celula utilizando tecnicas de ingenieria recombinante. Por ejemplo, un gen que estaba presente en la celula cuando la celula se aislo originalmente de la naturaleza. Un gen todavia se considera endogeno si las secuencias de control, tales como secuencias promotoras o potenciadoras que activan la transcripcion o la traduccion, se han alterado a traves de tecnicas recombinantes. A la inversa, el termino "heterologo" tambien se utiliza en el presente documento, y se refiere a una proteina o a un polinucleotido que no se produce de manera natural en una celula hospedadora.

The term "endogenous protein" refers to a protein that is native to a cell, or that is produced in it naturally. An "endogenous polynucleotide" refers to a polynucleotide that is in the cell and was not introduced into the cell using recombinant engineering techniques. For example, a gene that was present in the cell when the cell was originally isolated from nature. A gene is still considered endogenous if control sequences, such as promoter or enhancer sequences that activate transcription or translation, have been altered through recombinant techniques. Conversely, the term "heterologous" is also used herein, and refers to a protein or polynucleotide that does not occur naturally in a host cell.

La expresion "recombinacion homologa" se refiere al intercambio de fragmentos de ADN entre dos moleculas de ADN o cromosomas emparejados en sitios de secuencias de nucleotidos identicas o casi identicas. En determinados aspectos, la integracion cromosomica es una recombinacion homologa.

The term "homologous recombination" refers to the exchange of DNA fragments between two DNA molecules or paired chromosomes at identical or almost identical nucleotide sequence sites. In certain aspects, chromosomal integration is a homologous recombination.

La expresion "secuencias homologas", como se usa en el presente documento, se refiere a una secuencia de polinucleotidos o polipeptidos que tiene, por ejemplo, aproximadamente 100%, aproximadamente 99% o mas, aproximadamente 98 % o mas, aproximadamente 97 % o mas, aproximadamente 96 % o mas, aproximadamente 95 % o mas, aproximadamente 94 % o mas, aproximadamente 93 % o mas, aproximadamente 92 % o mas, aproximadamente 91 % o mas, aproximadamente 90 % o mas, aproximadamente 88 % o mas, aproximadamente 85 % o mas, aproximadamente 80 % o mas, aproximadamente 75 % o mas, aproximadamente 70 % o mas, aproximadamente 65 % o mas, aproximadamente 60 % o mas, aproximadamente 55 % o mas, aproximadamente 50 % o mas, aproximadamente 45 % o mas, o aproximadamente 40 % o mas de identidad de secuencia con otra secuencia de polinucleotidos o polipeptidos cuando se alinean de manera optima para la comparacion. En aspectos particulares, las secuencias homologas pueden conservar el mismo tipo y/o nivel de una actividad de interes particular. En algunos aspectos, las secuencias homologas tienen entre 85% y 100% de identidad de secuencia, mientras que en otros aspectos hay una identidad de secuencia de entre 90 % y 100 %. En aspectos particulares, hay una identidad de secuencia de 95 % y de 100 %.

The term "homologous sequences", as used herein, refers to a polynucleotide or polypeptide sequence having, for example, about 100%, about 99% or more, about 98% or more, about 97% or more, about 96% or more, about 95% or more, about 94% or more, about 93% or more, about 92% or more, about 91% or more, about 90% or more, about 88% or more, approximately 85% or more, approximately 80% or more, approximately 75% or more, approximately 70% or more, approximately 65% or more, approximately 60% or more, approximately 55% or more, approximately 50% or more, approximately 45 % or more, or about 40% or more sequence identity with another polynucleotide or polypeptide sequence when optimally aligned for comparison. In particular aspects, homologous sequences can retain the same type and / or level of an activity of particular interest. In some aspects, homologous sequences have between 85% and 100% sequence identity, while in other aspects there is a sequence identity between 90% and 100%. In particular aspects, there is a sequence identity of 95% and 100%.

"Homologia" se refiere a similitud de secuencia o identidad de secuencia. La homologia se determina utilizando tecnicas estandar conocidas en la materia (vease, por ejemplo, Smith y Waterman, Adv. Appl. Math., 2:482, 1981; Needleman y Wunsch, J. Mol. Biol., 48:443, 1970; Pearson y Lipman, Proc. Natl. Acad. Sci. USA 85:2444, 1988; programas tales como GAP, BESTFIT, FASTA y TFASTA en el paquete informatico de Wisconsin Genetics (Genetics Computer Group, Madison, WI); y Devereux et al., Nucl. Acid Res., 12:387-395, 1984). Un ejemplo no limitativo incluye el uso del programa BLASt (Altschul et al., Gapped BLAST y PSI-BLAST: a new generation of protein database search programs, Nucleic Acids Res. 25: 3389-3402, 1997) para identificar secuencias que puede decirse que son "homologas". Una version reciente, tal como las versiones 2.2.16, 2.2.17, 2.2.18, 2.2.19, o la ultima version, incluyendo subprogramas tales como blastp para comparaciones entre proteinas, blastn para comparaciones entre nucleotidos, tblastn para comparaciones entre proteinas y nucleotidos o blastx para comparaciones entre nucleotidos y proteinas, y con los siguientes parametros: numero maximo de secuencias devueltas 10000 o 100000; valor E (valor esperado) de 1e-2 o 1e-5, tamano de palabra de 3, matriz de puntuacion BLOSUM62, existencia de coste por hueco 11, extension de coste por hueco 1, puede ser adecuada. Un valor E de 1e-5, por ejemplo, indica que la probabilidad de que se produzca una coincidencia homologa al azar es de aproximadamente 1 en 10000, indicando de este modo una confianza alta de homologia verdadera.

"Homology" refers to sequence similarity or sequence identity. Homology is determined using standard techniques known in the art (see, for example, Smith and Waterman, Adv. Appl. Math., 2: 482, 1981; Needleman and Wunsch, J. Mol. Biol., 48: 443, 1970 ; Pearson and Lipman, Proc. Natl. Acad. Sci. USA 85: 2444, 1988; programs such as GAP, BESTFIT, FASTA and TFASTA in the Wisconsin Genetics software package (Genetics Computer Group, Madison, WI); and Devereux et al., Nucl. Acid Res., 12: 387-395, 1984). A non-limiting example includes the use of the BLASt program (Altschul et al., Gapped BLAST and PSI-BLAST: a new generation of protein database search programs, Nucleic Acids Res. 25: 3389-3402, 1997) to identify sequences that can be said They are "homologous." A recent version, such as versions 2.2.16, 2.2.17, 2.2.18, 2.2.19, or the latest version, including subprograms such as blastp for protein comparisons, blastn for nucleotide comparisons, tblastn for protein comparisons and nucleotides or blastx for comparisons between nucleotides and proteins, and with the following parameters: maximum number of sequences returned 10,000 or 100,000; E value (expected value) of 1e-2 or 1e-5, word size of 3, BLOSUM62 score matrix, existence of cost per hole 11, cost extension per hole 1, may be appropriate. An E value of 1e-5, for example, indicates that the probability of a random homologous coincidence is approximately 1 in 10,000, thus indicating a high confidence of true homology.

Las expresiones "cepa hospedadora" o "celula hospedadora" se refieren a un hospedador adecuado para un vector de expresion que comprende un ADN de la presente divulgacion.

The terms "host strain" or "host cell" refer to a suitable host for an expression vector comprising a DNA of the present disclosure.

C o m o se sabe en la m a te ria , el te rm in o "h ib rid a c io n " se re fie re al p ro ceso m ed ia n te el cua l una cadena de p o lin u c le o tid o s se une con una ca d e n a co m p le m e n ta r ia m ed ian te e m p a re ja m ie n to de bases. S e co n s id e ra que una se cu e n c ia de po lin u c le o tid o s se "h ib rid a s e le c tiva m e n te " con una secu en c ia de po lin u c le o tid o s de re fe renc ia , si las dos se cu e n c ia s se h ib ridan esp e c ifica m e n te en tre si en con d ic io n e s de h ib rid ac ion y lavado de rigu ros idad m o d e ra d a a alta . Las co n d ic io n e s de h ib rid ac ion d e p e nd en de la te m p e ra tu ra de fus io n (Tm) de l co m p le jo o so n d a de un ion al p o lin uc leo tido . P o r e jem p lo , no rm a lm e n te se p roduce una "r ig u ro s id a d m ax im a" a una te m p e ra tu ra de fus ion , Tm, de a p ro x im a d a m e n te -5 °C (5 °C po r de ba jo de la Tm de la sonda ); una "r ig u ro s id a d a lta " a a p ro x im a d a m e n te 5 -10 °C po r de ba jo de la Tm; una "r ig u ro s id a d in te rm e d ia " a a p ro x im a d a m e n te 10 -20 °C por de ba jo de la Tm de la sonda ; y una "r ig u ro s id a d ba ja" a a p ro x im a d a m e n te 20 -25 °C po r de ba jo de la Tm. F u n c io n a lm e n te , para id e n tific a r se cu e n c ia s que tien en id en tidad es tric ta o id en tidad casi es tr ic ta con la son da de h ib rid ac ion , pu ed en u tiliza rse co n d ic io n e s de rig u ros ida d m axim a; m ie n tras que para id e n tifica r o d e te c ta r h o m o logo s de se cu e n c ia s de po lin uc leo tidos , pueden u tiliza rse con d ic io n e s de h ib ridac ion con una rig u ros ida d in te rm ed ia o ba ja . Las con d ic io n e s de h ib rid ac ion con rig u ro s id a d m o d e ra d a y a lta son bien c o n o c id a s en la te cn ica . Un e jem p lo de co n d ic io n e s de rig u ros ida d a lta in c luye una h ib rid ac ion a a p ro x im a d a m e n te 42 °C en fo rm a m id a al 50 %, s Sc 5x , so lu c io n de D e n h a rd t 5X , S D S al 0 ,5 % y 100 pg /m l de A D N p o rta d o r d e sn a tu ra liza d o , se g u id o de la vad o dos ve ce s en 2 X S S C y S D S al 0 ,5 % a te m p e ra tu ra am b ie n te y dos vece s m as en 0 ,1 X S S C y S D S al 0 ,5 % a 42 °C. C om o e jem p lo de con d ic io n e s de rig u ros ida d m od e ra da se inc luye una in cub ac io n du ran te la noche a 37 °C en una so luc io n que com p ren de fo rm a m id a al 20 % , 5X S S C (N a C l 150 m M , c itra to tr iso d ico 15 m M ), fo s fa to de sod io 50 m M (pH 7,6), so luc io n de D e n h a rd t 5X, su lfa to de d e x tra n o al 10 % y 20 m g /m l de A D N de esp e rm a de sa lm on co rta d o y d e sn a tu ra liza d o , se g u id o de la vad o de los filtros en 1X S S C a una te m p e ra tu ra de a p ro x im a d a m e n te 37 °C a a p ro x im a d a m e n te 50 °C. Los exp e rtos en la m a te ria saben com o a ju s ta r la te m p e ra tu ra , la fu e rza ion ica y o tras con d ic ion es , segun sea necesa rio , para a d e c u a r fa c to re s ta le s com o la long itud de la so n d a y s im ila res.

As it is known in the ma ria, rm in o "h ib rid ac io n" refers to the most important process in which a chain of links is used. Unite with a co-channel channel, you will also like to base yourself. I know that one of the leaders of the group was "h ib rid ase le c tiva ment" with a section of the po l u u le le tt os de re fe reluctantly, if both of you are aware of this, you will have a special spirit in each other in conditions of disinfection and washing of moderate high standards. The co d nio n e s of h ib rid ac tion of the merger (Tm) of the co m p lem or so of an ion at the same time. P o jem p lo, no rm a lm in which a "r ig u ro s id adm ax im a" is produced by your merger, Tm, of ap ro x im adamen te -5 ° C (5 ° C below the Tm of the probe); a "r ig u ro s id a d a lta" a a p ro x im a d a m e n t 5 -10 ° C after the Tm; a "r ig u ro s id a d in te rm e d ia" a a p ro x im a d a m e n te 10 -20 ° C below the Tm of the probe; and a "r ig u ro s id a d ba ja" a a p ro x im a d a m e n t 20 -25 ° C po under the Tm. F unc io na lm en te, in order to identify, it is known that the identity is trite or almost identical with the sound of the ridiculous sound, because Use of coarse conditions for a maximum period; However, in order to identify the surrounding areas in which they are responsible for readings, they can be used with hybridization guidelines with a rigid stripe. d in te rm ed ia o ba ja. The con d ic io n e s of h ib rid ac ion with rig u ro s id a d m o d e ra d a y lta are very good in the technical field. An example of rigging conditions in which there is an average rate of 42 ° C in the form of 50%, Sc 5x, On the basis of D enhancer t 5X, 0.5% SDS and 100 pg / ml of DNA from the right side of your brain, it is guided by the vad or two times in 2 XSSC and SDS at 0.5% longer than two times and twice more at 0.1 XSSC and SDS at 0.5% at 42 ° C. C om oe jem for the conditions of rigors of the day, there is an in fl ower in the night at 37 ° C in a solution that includes of 20% fo rm am id, 5X SSC (N to C l 150 mM, it is to be used in 15 mM), 50 mM sodium fo (fa 7.6), In the case of D enha rd t 5X, its value of 10% dextra and 20 mg / ml of DNA from Spain is co-ordinated by your plan, it is indicated or from the filter or in the 1X SSC filters at a temperature of 37 ° C to 37 ° C to 50 ° C. The experts in the ma- ria know how comfortable you are, the fu e rza ica ica, after con, is, as necessary, to adapt easily. le s com o la long itud de la so ndays im ila res.

El te rm in o "h id ro ca rb u ro " se re fie re a co m p u e s to s qu im icos que con tienen los e lem e n tos ca rb on o (C ) e h id rogeno (H). T o d o s los h id ro ca rb u ro s con s tan de un e sq u e le to de ca rb o n o y de a to m o s de h id ro g e n o un id os a ese esque le to . A lg u n a s vece s , el te rm in o se u tiliza com o una fo rm a a b re v ia d a de la e xp res ion "h id ro c a rb u ro a lifa tico ". B a s ica m e n te hay tres tip o s de h id ro ca rbu ros : (1) h id ro ca rb u ro s a ro m atico s , que tien en al m en os a p ro x im a d a m e n te un an illo a ro m a tico ; (2) h id ro ca rbu ros sa tu rados , tam b ien con oc idos com o a lcanos, que ca recen de en laces dob les, tr ip le s o a ro m a tico s ; y (3) h id ro ca rb u ro s in sa tu ra dos , que en tre los a to m o s de ca rb on o tien en uno o m as en la ces do b les o tr ip le s e inc luyen , po r e jem p lo , a lqu eno s (p o r e jem p lo , d ien os ) y a lqu inos.

The rm in o "h id ro ca rb u ro" is referred to the chemicals that have the elements of the caRb on o (C) and the id rh (r). T o d o s the h id ro ca rb u ro s with s so of a sq e e of u r e r o u o s of m o s of h id ro g e n o an id es to that scheme. Sometime later, the tea is used as a form of the "h id ro c a rb u ro a ro a r a u r a u r a u r a u r a u r a u" a lifelic ". Basically, there are three types of ideas: (1) You have a lot of attics, which have a minimum of one year. robotic; (2) h id ro ca rbu ros sa tu rados, also with common ears or to lcanos, which are lacking in double bonds, tr ip le s or a r m ro micas; and (3) h id ro ca rb u ro s in sa tu ra ra, which between one year or another in the ces do b les or tr ip are included, because re jem p lo, lqu eno s (pore jem p lo, dien os) and lqu inos.

El te rm in o "iden ticos", en el con tex to de dos se cu e n c ia s de p o lin u c le o tid o s o po lip ep tido s , s ig n ifica que cua nd o las dos se cu e n c ia s se a linean pa ra o b te n e r una co rre sp o n d e n c ia m axim a , los res tos son igua les, ta l y com o se m ide u tiliza n d o un a lg o ritm o de an a lis is o co m p a ra c io n de se cu e n c ia s com o los d e sc rito s en este do cum e n to . P or e jem p lo , si cua nd o se a linean co rre c ta m en te , los se g m e n to s c o rre sp o n d ie n te s de dos se cu e n c ia s tie n e n res tos id e n tico s en 5 po s ic io ne s de 10, se d ice que las dos se cu e n c ia s tie n e n una id en tidad de l 50 %. La m ayo ria de los p ro g ra m a s b io in fo rm a tico s in d ica n el po rcen ta je de id en tidad sob re las reg io nes de se cu e n c ia a line ad as , que n o rm a lm e n te no son las m o le cu la s com p le ta s . Si un a lin e a m ie n to es lo s u fic ie n te m e n te la rgo y con tiene su fic ie n te s res tos iden ticos, se pued e ca lc u la r un v a lo r de exp ec ta tiva , que ind ica que es im p roba b le que el n ive l de id en tidad en el a lin e a m ie n to ocu rra po r azar.

The rm in or "identical" tea, in the context of two, it is a matter of being able to understand the meaning of each other, it means that when the two are If they are aligned to obtain a maximum distribution, the remainders are equal, as well as as a year or rhythm of annual analysis. This is the case of those of the rites in this process. For example, if they are aligned with each other, the reports of the two of them are the same as those with identical identities. in 5 cases of 10, it was said that both of them had an identity of 50%. The majority of the most important in-house programs in the field of identity in relation to the sectoral regions that line them, which does not They're not the best ones in the world. If a year is the only thing that tells you about it and with it it has its own ideals, then it can be explained , which indicates that it is important that the level of identity in the foreign sector is random.

Las e xp res ione s "m u ta c ion de m e jo ra " o "m u ta c io n de po tenc iac io n del ren d im ie n to " se re fie ren a una m u tac ion en una p ro te ina que co n d u ce a p ro p ieda de s a lte rad as , que con fie ren un re n d im ie n to m e jo ra d o en te rm in o s de una d ian a y /o p ro p ieda d de sea da de una p ro te ina en c o m p a ra c io n con una p ro te ina p recu rso ra .

The "best improvement" or "great power" models refer to a mu- tion in one way. I would like you to know about the radius, that with faith I will give you a better income in a few days a day and / or a day of that day. of a person compared to a person.

El te rm in o "in se rc io n " cu a n d o se usa en el co n te x to de una se cu e n c ia de po lip ep tido s , se re fie re a una inse rc io n en la se cu e n c ia de a m in o a c id o s de un po lip e p tid o p recu rso r, da ndo com o resu lta d o un po lip e p tid o m u tan te que t ie n e un a m in o a c id o que se in se rta e n tre dos a m in o a c id o s co n tig u o s e x is te n tes , es decir, res tos de a m in o a c id o s adyacen tes , que estan p re sen te s en el p o lip ep tido p recurso r. El te rm in o "inse rc ion ", cua nd o se u tiliza en el con tex to de una se cu e n c ia de po lin uc leo tidos , se re fie re a una in se rc ion de uno o m as nu c le o tido s en el p o lin uc leo tido p re cu rso r en tre dos nu c le o tid o s con tigu os ex is te n tes , es decir, nu c le o tido s ad yace n tes , que estan p re sen te s en los p o lin u c le o tid o s p recu rso res .

The rm in o "in se rc io n" when used in the context of a lipid episodes, refers to an insertion in the sect. On the other hand of a single lipid, it is common to find a small number of people so that you have an amne or an id which is inserted between two years or two years ago, that is, the rest of the countries or adjacent territories, which are present in the area lip ep tido p resource r. The rm in or "inse rtion", when used in the context of one of the most important reports, refers to an inception of one or more nuances. C o t tido s in the po lin uc le t tido p re cu rso r in two nu c le tid os with tiguos ex es tents, that is, nu c le tido s ad ad lying, which They are present in the po l uc le or tid osp rec rso res.

El te rm in o "in tro d u c id o ", en el con tex to de in tro d u c ir una secu en c ia de p o lin u c le o tid o s en una ce lu la , se re fie re a cu a lq u ie r m e tod o a d e cu a d o para tra n s fe r ir la secu en c ia de po lin u c le o tid o s a la ce lu la . D ichos m e tod os de in tro d u cc io n in c luyen , pero sin lim itac ion , fus io n de p ro to p las tos , tra n s fe cc io n , tra n s fo rm a c io n , con ju g a c io n y tra n sd u cc io n (vease, po r e jem p lo , Fe rra ri e t al., G en e tics , en H a rdw o od e t al, (eds.), B ac illu s , P le nu m P ub lish ing C orp., pags. 57 -72 , 1989).

The rm in or "in tro duc id o" tea, in the context of in tro duc go a section of the po l u u tid os in a ce lu la, is referred to cu l lq In other words, for the sake of faith, go to the police department. These methods are included in the process, but without limitation, merger of coughs, through faith, through fo rm ac tion, with J ga ga io n and ntra n sd u cc io (see, po re jem p lo, Fe rra ri et al., G in e tics, in H a rdw or od et al, (eds.), B ac illu s , P le nu m P ub lish ing C orp., Pp. 57-72, 1989).

Los te rm in o s "a is la d o " o "p u rifica d o " s ign ifican un m a te ria l que se e lim in a de su en to rno o rig ina l, po r e jem p lo , el e n to rn o na tu ra l en caso de que sea de o rige n na tu ra l, o un ca ldo de fe rm e n ta c io n si se p roduce en un m e d io de fe rm e n ta c io n de ce lu la s ho sp e d a d o ra s reco m b in an te s . S e d ice que un m a te ria l esta "p u rif ica d o " cuando esta p re sen te en una c o m p o s ic io n p a rtic u la r en una c o n ce n tra c io n m a y o r o m e n o r que la c o n c e n tra c io n que ex is te an tes de re a liza r la e ta pa o e ta pas de pu rifica c ion . P o r e jem p lo , con resp ec to a una com p o s ic io n que ha b itu a lm e n te se e n cu e n tra en un o rg a n ism o na tu ra l o de t ip o s ilves tre , d icha co m p o s ic io n se "p u rif ica " cu a n d o la co m p o s ic io n fina l no in c luye a lgun m a te ria l de la m a triz o rig ina l. C o m o otro e jem p lo , dond e una com p o s ic io n se e n cu e n tra en combinacion con otros componentes en un medio de fermentacion de celulas hospedadoras recombinantes, esa composicion se purifica cuando el medio de fermentacion se trata de una manera que se elimine algun componente de la fermentacion, por ejemplo, desechos celulares u otros productos de fermentacion, a traves de, por ejemplo, centrifugacion o destilacion. Como otro ejemplo, un polinucleotido o polipeptido natural presente en un animal vivo no esta aislado, pero el mismo polinucleotido o polipeptido, separado de algunos o de todos los materiales coexistentes en el sistema natural, esta aislado, si dicho proceso se realiza a traves de ingenieria genetica o separacion mecanica. Dichos polinucleotidos pueden ser partes de vectores. Como alternativa, dichos polinucleotidos o polipeptidos pueden ser partes de composiciones. Dichos polinucleotidos o polipeptidos pueden considerarse "aislados" porque los vectores o composiciones que los comprenden no forman parte de sus entornos naturales. En otro ejemplo, se dice que un polinucleotido o proteina se purifica si da lugar basicamente a una banda en un gel o transferencia de electroforesis.

The te rm in os "a is la do" or "pu rifica do" s ignify a material that is limited to its environment or rigth, as far as possible. rn o na tu ra l in the event that it is of or governs na ra ra, or a ca rdo of faith rm in ta if it is produced in a measure of faith rm in ta c io n de ce lu la s ho sp edado ra s reco mb in an te s. I know that a mother is "pu rif ica do" when she is present in a mainstream composition in a major convention with the greatest concentration that existed. IS BEFORE YOU REPRESENT THE COUNCIL OF PURIFICATION. P o je je p p, with respect to a com- position that has come to me in which you find yourself in one of the most important aspects of your interest, It is "pu rif ica" when the company is fine in some way in the middle of the matrix. Like another jem p lo, dond e a com posi io n en cu en tra en Combined with other components in a recombinant host cell fermentation medium, that composition is purified when the fermentation medium is treated in a manner that removes some component of the fermentation, for example, cellular debris or other fermentation products, through of, for example, centrifugation or distillation. As another example, a natural polynucleotide or polypeptide present in a live animal is not isolated, but the same polynucleotide or polypeptide, separated from some or all of the coexisting materials in the natural system, is isolated, if said process is performed through genetic engineering or mechanical separation. Such polynucleotides can be parts of vectors. Alternatively, said polynucleotides or polypeptides may be parts of compositions. Such polynucleotides or polypeptides can be considered "isolated" because the vectors or compositions comprising them are not part of their natural environments. In another example, it is said that a polynucleotide or protein is purified if it basically results in a band in a gel or electrophoresis transfer.

El termino "madura", en el contexto de una proteina, significa una forma de una proteina o peptido que se encuentra en su forma funcional final. Para ejemplificar, una forma madura de una tioesterasa de la presente divulgacion comprende los restos de aminoacidos 1-182 de la SEC ID NO: 31 expuesta en la FIG. 58.

The term "mature", in the context of a protein, means a form of a protein or peptide that is in its final functional form. To exemplify, a mature form of a thioesterase of the present disclosure comprises amino acid residues 1-182 of SEQ ID NO: 31 set forth in FIG. 58.

La expresion "derivados de acidos grasos modificados" se refiere a productos fabricados, al menos en parte, a partir de una parte de la ruta biosintetica de acidos grasos de una celula hospedadora recombinante, en donde el producto difiere del producto fabricado por dicha celula hospedadora en ausencia de la tioesterasa mutante de la divulgacion. Por tanto, cuando se introduce una tioesterasa mutante (o su equivalente de origen natural) en una celula hospedadora recombinante, se da como resultado la produccion de un derivado de acido graso que tiene un perfil de producto diferente, por ejemplo, una concentracion mas alta o mas baja de ciertos derivados de acidos grasos que tienen una longitud de cadena especifica, o una concentracion mas alta o mas baja de un cierto tipo de derivado de acidos grasos, ese material de acido graso esta "modificado" en el contexto de esta divulgacion.

The term "modified fatty acid derivatives" refers to products manufactured, at least in part, from a part of the biosynthetic fatty acid pathway of a recombinant host cell, wherein the product differs from the product manufactured by said host cell. in the absence of the thioesterase mutant of the disclosure. Therefore, when a mutant thioesterase (or its equivalent of natural origin) is introduced into a recombinant host cell, the production of a fatty acid derivative having a different product profile, for example, a higher concentration, results or lower of certain fatty acid derivatives having a specific chain length, or a higher or lower concentration of a certain type of fatty acid derivative, that fatty acid material is "modified" in the context of this disclosure .

Las expresiones "tioesterasa mutante" o "tioesterasa variante", se refieren a una tioesterasa que comprende una mutacion con referencia a una tioesterasa precursora.

The terms "mutant thioesterase" or "variant thioesterase" refer to a thioesterase comprising a mutation with reference to a precursor thioesterase.

El termino "mutacion", en el contexto de una proteina, se refiere a una modificacion de la secuencia de polinucleotidos que produce un cambio en la secuencia de un polinucleotido con referencia a una secuencia de polinucleotidos precursora. Una secuencia de polinucleotidos mutante puede referirse a una alteracion que no cambia la secuencia de aminoacidos codificada, por ejemplo, con respecto a la optimizacion de codones con fines de expresion, o que modifica un codon de tal manera que produce una modificacion de la secuencia de aminoacidos codificada. Las mutaciones pueden introducirse en un polinucleotido a traves de cualquiera de los diversos metodos conocidos por los expertos en la materia, incluyendo mutagenesis aleatoria, mutagenesis especifica de sitio, mutagenesis dirigida por oligonucleotido, transposicion de genes, tecnicas de evolucion dirigida, mutagenesis combinatoria, mutagenesis de saturacion de sitios, entre otros.

The term "mutation", in the context of a protein, refers to a modification of the polynucleotide sequence that produces a change in the sequence of a polynucleotide with reference to a precursor polynucleotide sequence. A mutant polynucleotide sequence may refer to an alteration that does not change the encoded amino acid sequence, for example, with respect to codon optimization for expression purposes, or that modifies a codon in such a way that it produces a modification of the sequence of encoded amino acids Mutations can be introduced into a polynucleotide through any of the various methods known to those skilled in the art, including random mutagenesis, site-specific mutagenesis, oligonucleotide-directed mutagenesis, gene transposition, directed evolution techniques, combinatorial mutagenesis, mutagenesis. of saturation of sites, among others.

"Mutacion" o "mutada" significa, en el contexto de una proteina, una modificacion en la secuencia de aminoacidos que produce un cambio en la secuencia de una proteina con referencia a una secuencia de proteina precursora. Una mutacion puede referirse a una sustitucion de un aminoacido por otro aminoacido, a una insercion o a una delecion de uno o mas restos de aminoacidos. Especificamente, una mutacion tambien puede ser el reemplazo de un aminoacido con un aminoacido no natural, o con un aminoacido modificado quimicamente o restos similares. Una mutacion tambien puede ser un truncamiento (por ejemplo, una delecion o interrupcion) en una secuencia o una subsecuencia de la secuencia precursora. Una mutacion tambien puede ser una adicion de una subsecuencia (por ejemplo, dos o mas aminoacidos en un tramo, que se insertan entre dos aminoacidos contiguos en una secuencia proteica precursora) dentro de una proteina, o en cualquier extremo terminal de una proteina, aumentando asi la longitud de la proteina (o alargandola). Se puede hacer una mutacion modificando la secuencia de ADN correspondiente en la proteina precursora. Las mutaciones pueden introducirse en una secuencia de proteinas mediante metodos conocidos en la materia, por ejemplo, creando secuencias sinteticas de ADN que codifican la mutacion con referencia a proteinas precursoras, o alterando quimicamente la propia proteina. Una proteina "mutante", como se usa en el presente documento, es una proteina que comprende una mutacion. Por ejemplo, tambien es posible crear un mutante, reemplazando una parte de una tioesterasa con una secuencia de tipo silvestre que corresponde a dicha parte, pero que incluye una variacion deseada en una posicion especifica que se produce de manera natural en la secuencia de tipo silvestre.

"Mutation" or "mutated" means, in the context of a protein, a modification in the amino acid sequence that produces a change in the sequence of a protein with reference to a precursor protein sequence. A mutation may refer to a substitution of one amino acid for another amino acid, an insertion or a deletion of one or more amino acid residues. Specifically, a mutation can also be the replacement of an amino acid with an unnatural amino acid, or with a chemically modified amino acid or similar moieties. A mutation can also be a truncation (for example, a deletion or interruption) in a sequence or a sub-sequence of the precursor sequence. A mutation can also be an addition of a sub-sequence (for example, two or more amino acids in a stretch, which are inserted between two contiguous amino acids in a precursor protein sequence) within a protein, or at any terminal end of a protein, increasing thus the length of the protein (or lengthening it). A mutation can be made by modifying the corresponding DNA sequence in the precursor protein. Mutations can be introduced into a protein sequence by methods known in the art, for example, by creating synthetic DNA sequences that encode the mutation with reference to precursor proteins, or by chemically altering the protein itself. A "mutant" protein, as used herein, is a protein that comprises a mutation. For example, it is also possible to create a mutant, replacing a part of a thioesterase with a wild-type sequence that corresponds to that part, but that includes a desired variation in a specific position that occurs naturally in the wild-type sequence. .

Un "equivalente natural", en el contexto de la presente invencion, se refiere a una tioesterasa natural, o a una parte de la misma, que comprende un resto natural, en donde el resto natural corresponde a una mutacion en 'TesA (por ejemplo, una mutacion en la SEC ID NO: 31 de la FIG. 58) que ha introducido una propiedad alterada deseable para 'TesA. En la FIG. 55 se proporcionan ejemplos de tioesterasas equivalentes naturales que tienen dichas modificaciones.

A "natural equivalent", in the context of the present invention, refers to a natural thioesterase, or a part thereof, which comprises a natural moiety, wherein the natural moiety corresponds to a mutation in 'TesA (for example, a mutation in SEQ ID NO: 31 of FIG. 58) that has introduced a desirable altered property for 'TesA. In FIG. Examples of natural equivalent thioesterases having such modifications are provided.

La expresion "unido operativamente", en el contexto de una secuencia de polinucleotidos, se refiere a la colocacion de una secuencia de polinucleotidos en una relacion funcional con otra secuencia de polinucleotidos. Por ejemplo, un ADN que codifica un lider de secrecion (por ejemplo, un peptido senal) esta unido operativamente a un a Dn que codifica un polipeptido si se expresa como una preproteina que participa en la secrecion del polipeptido. Un promotor o p o te n c ia d o r es ta un ido o p e ra tiva m e n te a una s e cu e n c ia c o d ifica n te si a fe c ta a la tra n s c r ip c io n de la secu en c ia . Un s itio de un ion al rib oso m a esta un ido o p e ra tiva m e n te a una se cu e n c ia co d ifica n te si es ta c o lo ca d o de m a n e ra que fa c ilite la tra d u cc io n . En genera l, "u n id o o p e ra tiva m e n te " s ig n ifica que las se cu e n c ia s de A D N que estan un idas son co n tig u a s y, en el caso de un lide r de secre c io n , co n tig u o y en el m ism o m arco de lectura.

The term "operably linked", in the context of a polynucleotide sequence, refers to the placement of a polynucleotide sequence in a functional relationship with another polynucleotide sequence. For example, a DNA encoding a secretion leader (for example, a signal peptide) is operably linked to a Dn encoding a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide. A promoter It is an operational operation that has been coded by a coding code if it is up to date to the cross-section of the section. One year from an ion to the mainland is an operational one to a coded sign if it is as well as possible to facilitate the work of the work. In general, "a positive operating id" means that the DNA signals that are attached are co-tious, in the case of a secrecy leader, co n tig uoy in the same reading arc.

La e xp re s io n "re g io n o p e ro n " se re fie re a un g ru p o de ge ne s co n tig u o s que se tra n s c r ib e n com o una so la un id ad de tra n sc r ip c io n a p a rtir de un p ro m o to r com un, y po r lo ta n to su je to a reg u lac io n con jun ta . En a lgunos aspec tos , el op e ron in c luye un gen regu lado r.

The e xp re s io n "re g io nope ro n" refers to a group of genes that were written in common with only one trad id of tra n sc r ip c io napa rtir of a p ro mo to r common, and that is why your region is together. In some respects, the op e ron in c has a regu side gene r.

La e xp res ion "a lin e a m ie n to op tim o", se re fie re al a lin e a m ie n to que da la pun tua c io n de a lin e a m ie n to g lob a l m as alta. El te rm in o "o rto lo g o s " o la exp res ion "g e n e s o rto log os", se re fie ren a ge ne s en d ife re n te s esp e c ie s que han e v o lu c io n a d o a p a rtir de un gen an ce s tra l com un po r esp ec iac io n . N o rm a lm e n te , los o rto lo g o s con se rva n la m ism a fu n c io n d u ra n te e l tra n s c u rs o de la e vo lu c io n . La id e n tifica c io n de o rto lo g o s e n cu e n tra uso en la p re d icc io n f ia b le de la fun c ion gen ica en g e n o m a s rec ien se cu en c iad os .

The expression "a lin e a m ie n to op tim o", refers to the line that gives the highest point of the line. The rm in o "o rto lo gos" or the "geneo rto log os" expression refers to the people in which they have evolved. of a gene that is common to the community. N o rm a lm e n te, the o rto lo g o s con rva n la m ism a fu n c io n d u ra n te e l n th e c u rs o of the e vo lu c io n. The identification of the o rto lo g o s e n when it was used in the p r d d icc io b le of the gene function as a new one in recent years.

La "s o b re e x p re s io n " de una ce lu la h o sp e d a d o ra o una ce lu la h o sp e d a d o ra "so b re xp re sa d a ", se p ro du ce si la e n z im a se exp resa en la ce lu la a un n ive l m as a lto que el n ive l en dond e se exp resa en una ce lu la de t ip o s ilves tre co rre spo nd ien te .

The "sob re exp re s io n" of a ce la ho sp edado ra or a ce lu la ho sp edado ra "so b re xp re sa da", if the enzyme is expressed resa in the city to a level lm more than the level in which it is expressed in a city of the same time as the spouse.

El te rm in o "p a ra lo g o " o la exp res ion "g en es pa ra logo s" se re fie ren a genes que estan re la c io n a d o s po r du p licac ion d e n tro de un genom a. M ie n tras que los o rto lo g o s con se rva n la m ism a fu n c io n en el cu rso de la evo lu c ion , los p a ra lo g o s d e sa rro lla n nu eva s fu n c io n e s , a u n q u e a lg u n a s fu n c io n e s a m en ud o estan re la c io n a d a s con las o rig ina les . C o m o e je m p lo s de genes pa ra logo s se inc luyen , pero sin lim itac ion , genes que cod ifica n la m io g lo b ina y la he m og lob in a , que su rg en de l m ism o a n te p a sa d o a n tigu o pero que e vo lu c io n a ro n pa ra te n e r d ife ren te s fun c ion es . La exp res ion "co e fic ie n te de rep a rto " s ig n ifica la co n ce n tra c io n de e q u ilib rio de un co m p u e s to en una fa se o rg an ica d iv id ida en tre la co n ce n tra c io n en eq u ilib rio en una fase acu osa (p o r e jem p lo , en un ca ldo de fe rm e n ta c io n ). En un a sp e c to del s is tem a b ifa s ico de sc rito en el p re sen te docum e n to , la fase o rg an ica esta fo rm a d a po r el de riva do de ac ido g raso d u ran te el p roceso de p roducc ion . En de te rm in a d a s c ircu ns tan c ias , ta m b ie n puede p ro p o rc io n a rse una fase o rgan ica , po r e jem p lo , para fa c ilita r la sep a ra c ion de l p roduc to , puede p ro p o rc io n a rse una cap a de o c ta no al ca ldo de fe rm e n ta c io n . C u ando se de sc rib e un s is tem a de dos fases, el coe fic ie n te de reparto , P, g e n e ra lm e n te se expone en te rm in o s de logP. Un co m p u e s to con un logP de 1 te n d ria un rep a rto de 10: 1 en la fase o rgan ica . Un c o m p u e s to con un logP de -1 te n d ria un rep a rto de 1:10 en la fase o rgan ica . A l e le g ir un ca ldo de fe rm e n ta c io n y una fa se o rg a n ic a a d e cu a d o s , un d e riva d o de ac ido g ra so con un v a lo r logP a lto se se p a ra ra en la fa se o rg a n ica in c luso a c o n ce n tra c io n e s m uy ba jas en el rec ip ie n te de fe rm en tac io n .

The te rm in o "para lo go" or the expression "g en es para ra logo s" refers to genes that are linked to the placement of a genom a . M e n after the prayers have been carried out with the same moment in the course of the evolution, the gospel will save the number of evasions, Although some of them have been nested in you or are laughing with the leaders. How do you think about genes that include genes, but without limitation, genes that encode my own and that you have, that your rg is lm ism? Oan te pa sa doan tigu o but that e vo lu c io na ro n pa ra te nerd ife ren te s fun c ion es. The "co-fiction of a" term means the equi tional balance of a cost in a single phase of the id id between the conn tion in eq u ilio rio in an aqueous phase (for example, in a situation of faith in that period). In one year of the only science of science in the world, the phase or phase of the program is that of the private sector. P roduction p roduction. In the course of such an irration, such a phase or rganic phase may also be possible, for example, to facilitate the sep- aration of lp roduc However, there may be an occasional captain not to the end of faith in this situation. With a two-phase system, the distribution coefficient, P, g e n e ra lm e n, is presented in rm in or s of logP. A co-p u e s with a logP of 1 would give a 10: 1 ratio in the phase or organ. A c o m p u e s with a logP of -1 would not have a ratio of 1:10 in the phase or organ. You will have a time of faith in this situation and a family will be able to do so, one of the most important r ... It will be only in the same way as in the case of faith in the tac io n.

Las e x p re s io n e s "p o rce n ta je de id en tidad de se cu e n c ia " "p o rce n ta je (% ) de id en tidad de se cu e n c ia de am ino ac id os ", "p o rce n ta je de id en tidad de se cu e n c ia gen ica ", y /o "p o rce n ta je de id en tidad de se cu e n c ia de p o lin uc leo tidos ", con resp ec to a dos se cu e n c ia s de po lip ep tido s , p o lin u c le o tid o s y /o de genes (seg un co rre spo nda ), se re fie ren al po rcen ta je de restos que son iden ticos en las dos se cu e n c ia s cuando las s e cu e n c ia s estan a line ad as de m anera op tim a . P o r lo tan to , una id en tidad de se cu e n c ia de a m in o a c id o s de 80 % s ig n ifica que, en dos s e cu e n c ia s de po lip e p tid o s a lin e a d a s op tim am e n te , el 80 % de los a m in o a c id o s son iden ticos.

The "po rce n ta je of identity in the sense of security" "power of identity (%) of identity in the sense of love", "po rce n ta je of identity in the sense of genetics ", and / or" power of identity in the sense of security of the population ", with respect to two When it comes to some epitomial lipids, po u u le le tid osy / o of genes (according to sporadically), they refer to the potency of remains that are identical in both cases. When you know what they are, they are in line with each other. First of all, an identity of 80% or 80% means that, in two cases, it is possible to have a good idea of timeline. am en te, 80% of the am in o id are identical.

El te rm in o "p la sm id o " se re fie re a una con s tru cc io n de A D N b ica ten a rio (bc) c ircu la r que se u tiliza com o v e c to r de c lo na c ion , y que fo rm a un e lem e n to ge n e tico a u to rre p lica n te e x tra c ro m o s o m ic o en a lg u n o s e u ca rio ta s o p ro ca rio tas , o se in te g ra en el c ro m o so m a de l hospedador.

The rm in o "p la sm id o" refers to one with basic DNA DNA structure (bc) c ircu la r that is used as a national sheep This is the reason for the development of a common ethnic lem ture that will also be taken into account, or that it is intended g ra in the mo mo so ma of the host.

La e xp re s io n " tio e s te ra s a p re cu rso ra " se re fie re a una p ro te in a t io e s te ra s a de la que pu ede o b te n e rse la tio e s te ra s a m u tan te de la d ivu lg ac ion , a tra ve s de, po r e jem p lo , m ed io s re co m b in a n te s o qu im icos . Son e je m p lo s de t io e s te ra sa s p re cu rso ras las t io e s te ra sa s na tu ra les o de t ip o s ilves tre que p roceden de p lan tas, an im a le s o m ic ro b io s . U na tio e s te ra s a p re cu rso ra ta m b ie n pu e d e s e r una tio e s te ra s a no na tu ra l. Un e jem p lo de una tio e s te ra s a no na tu ra l es una t io e s te ra sa fab ricad a , po r e jem p lo , m ed ia n te m u ta c io n a lea to ria , s in tes is qu im ica , evo luc ion m o lecu la r, o m u ta g e n e s is d ir ig ida a s itio , que puede s e rv ir com o un pun to de pa rtida util desde el cua l d is e n a r y/o c re a r las tio e s te ra s a s m u tan tes de la d ivu lg ac ion .

The e xp re s io n "uncle es te ra sap re cu rso ra" refers to a p ro te in at io es te ra sa from which the uncle can get you is the sam sam so I d d u lg ac ation, through, p re jem p lo, m ed io s re co mb in an te so chemicals. They are the only thing about you, what is the reason that the people are the ones that are the ones that come from the streets, an im a le som ic ro b io s. U na uncle e s te ra s a p re cu rso ra ta m b ie n pu e d e s e r an uncle e s te ra s a no na tu ra l. An e jem p about an uncle is te ra sa not na tu ra l is an un tio un te ra sa fab ricad a, po re jem p lo, m ed ia n te mu ta c io na read to ria, s What is chemical? Evo luc ion mo lecu la r, omu ta g e nes is d go ig so asio, which can be used as a useful starting point since the fourth year. / oc re ar las unio es te ra sasmu tan te de la d ivu lg ac ion.

Un "ce b a d o r" es un o ligo nu c le o tido , p ro du c ido de m an e ra na tu ra l, co m o en una m ue s tra de d iges tion pu rifica da por res tricc ion , o p ro du c ido s in te ticam e n te , que es cap az de a c tu a r com o un pun to de in ic io de s in tes is cu a n d o se co loca en co n d ic io n e s en las que se induce la s in te s is de un p ro du c to de ex te ns io n c e b a d o r que es c o m p le m e n ta rio a una cad en a de p o lin u c le o tid o s de re fe renc ia . Las co n d ic io n e s a d ecu ad as inc luyen , po r e jem p lo , la p resenc ia de nu c le o tid o s y de un agen te in d u c to r ta l com o una A D N po lim e rasa , y una te m p e ra tu ra y un pH adecuados . P re fe re n te m e n te , para o b te n e r una e fica c ia m ax im a en la am p lifica c io n , un c e b a d o r es m on oca te na rio , pero, com o a lte rna tiva , pued e s e r b ica ten a rio . Si es b ica te n a rio (do b le cadena), un c e b a d o r pued e tra ta rs e p rim e ro para se p a ra r sus cad en as an tes de u tiliza rlo para p re p a ra r p ro du c to s de ex te ns io n . En asp ec tos pa rticu la res , un c e b a d o r es un o lig o d e so x irr ib o n u c le o tid o . En c ie rto s asp e c to s p re fe ridos , un c e b a d o r es lo s u fic ie n te m e n te la rgo co m o pa ra in ic ia r la s in te s is de p ro du c to s de ex te ns io n en p re sen c ia de un agen te induc to r. Las lo n g itu d e s exa c tas de los ce b a d o re s d e p e n d e ra n de d ive rso s fa c to re s , in c lu ye n d o la te m p e ra tu ra , la fu e n te de l ceb ad o r, y de los m e to d o s u tiliza d o s para la a m p lifica c io n .

A "primer" is one of the most important reasons, but also in one of the most important ways in which it can be derived, op ro du c gone s in te ticam en te, which is the cap of ac tu ar com o a point of in i of s in tes is where it is co-located in situations in which it is induced the in s you s is a p ro du to c tio ns of ex te ncebador is comp le men ta rio a cad in a lin uc po tid or you will re ia renc faith. The com nd ic io nesad ecu ad as inc show, po re jem p lo, the presence of a new agency that leads to a common DNA, and a you measure your ra and an adequate pH. P re fe re n te ment, in order to obtain a maximum efficiency in the am l p ict, a primer is only national, but, as a matter of fact, it can Eserbica have the river. If it is basic to you (do b le chain), a primer can be used to stop your body before using it for the first time. s ex ex te ns io n. In aspic cough, a primer is one of the two sources of irrigation. In this context, a primer is the only thing that is the same as the first step of the day. Not only in the presence of an agent. The exact slopes of the primaries It will depend on the number of people, in which way you will be able to measure it, the future of the program, and the methods will be used for the qualification. io n.

El te rm in o "so n d a " se re fie re a un o lig o n u c le o tid o , p ro d u c id o de m an e ra na tu ra l com o en una d iges tion p u rifica da po r res tricc ion o p ro du c ido de m an e ra s in te tica , re co m b in a n te o m ed ia n te am p lifica c io n po r PCR, que es capaz de h ib rid a rse con o tro o lig o n u c le o tid o de in te res. Una son da puede s e r m o n o ca te n a ria o b ica tenaria . Las sondas son u tiles en la de tecc io n , id e n tifica c io n y a is la m ie n to de s e cu e n c ia s g e ne ticas pa rticu la res . S e co n tem p la que c u a lq u ie r so n d a u tilizad a en la p re se n te d ivu lg a c io n se m arq ue con cu a lq u ie r "m o le cu la in d ica do ra ", po r lo que se ra de te c ta b le en cu a lq u ie r s is tem a de de tecc ion , inc luyendo , pero sin lim itac ion , s is te m a s e n z im a tico s (p o r e jem p lo , ensayos de t ip o E L IS A u o tros e n sayo s h is to q u im ico s ba sad os en enz im as), flu o re sce n te s , ra d io a c tivo s y lu m in iscen te s . No se p re te nd e lim ita r la p re sen te inven c ion a n ingun s is te m a o m arca je de de tecc io n pa rticu la r.

The rm in o "so nda" refers to a legal entity, but also in a common way in a public decision. Tr trickle op ro du c gone from m an e ra s in t tica, re co mb in an te om ed ia n t he p l p tio n t r r PCR, which is capable of h ib rid to go with another or lig onuc le or tid o of interest. A son s day can be basic or basic. The probes are useful in the tection, identification and identification method, and it is the most important method for the country. S e co n temp p la que lq u ie r so ndau tilizad a in p re se n te d ivu lg ac i n arq ue with what a lq u ie r "mo le cu la in d ica dica ra ", because what will be heard about the situation, including, but without limitation, is immantic mastery (for example) For example, t ip or EL IS essays in sayo sh is to chem s ba sad os in enzymes), flow or re sce n te s, the active day s and lu m in iscen te s. It is not possible to limit the invention to any way but also to the archeology of the country.

Un "h o s p e d a d o r de p ro du cc ion " es una ce lu la u tiliza d a pa ra p ro d u c ir p ro du c to s . T a l com o se d e sve la en el p re se n te do cum e n to , un h o sp e d a d o r de p ro du cc ion se m od ifica para e x p re s a r o s o b re e x p re s a r genes se lecc io na do s , o para te n e r una exp res ion a te nua da de genes se lecc io na do s . C o m o e je m p lo s no lim itan te s de ho sp e d a d o re s de p ro d u cc io n se in c luye n ce lu la s de p lan tas, an im a les , se re s hum anos, ba c te ria s , le vad u ras , c ia n o b a c te ria s , a lg a s y /o de h o ng os fila m en to sos .

A "h o s p e d a d o r of p ro du cc ion" is a use that can be used to go to the city. In the sense that in the country it is possible to do so, a person who is age-old is able to expire in order to express genes that were read, or to have an expiration to any of the genes was read na s s. How do you know what is wrong with you now? S p o du r s p ro du cc io n in c luye n ce lu la s de p lantas, an im a les, se re s hu years, ba I would laugh at you, I would laugh at you, I would like to be in line.

Un "p ro m o to r" es una secu en c ia de p o lin u c le o tid o s que fu n c io n a para d ir ig ir la tra n s c r ip c io n de un gen en d irecc ion 3'. En asp e c to s p re fe ridos, el p ro m o to r es ap rop iad o para la ce lu la h o spe da do ra en la que se exp resa el gen diana. El p rom oto r, ju n to con o tras se cu e n c ia s de p o lin u c le o tid o s reg u la d o ra s de la tra n sc r ip c io n y de la tradu cc io n (tam b ien de n o m in a d a s "se cu e n c ia s de con tro l"), es n e cesa rio para e xp re sa r un gen dado. En gene ra l, las s e cu e n c ia s reg u la d o ra s de la tra n sc r ip c io n y de la tra d u cc io n inc luyen , pero sin lim itac ion , se cu e n c ia s p rom oto ras, s itio s de un ion a ribosom as, se cu e n c ia s de in ic io y te rm in a c io n de la tra n sc rip c io n , s e cu e n c ia s de in ic io y te rm in a c io n de la tra d u cc io n y s e cu e n c ia s p o te n c ia d o ra s o ac tiva do ras .

A "p ro m o to r" is a section of p o lin u c le o tid s s that was able to go the route of a gene in direction 3 '. In asp e c to s p re fe r rides, the p r m o to r is suitable for the time in which the target gene is expressed. The p rum autumn, with or after, is the source of the regu la tors of the tra c sc r ip c io n and of the translation cc (tam b In the name of "they are aware of each other"), it is necessary for a given gene. In general, the registers of the tra n sc r ip c io n and of the tra du ctio inc include, but without limitation, it is en c After a ribosome ion, it is the beginning of the process and the process of the work is carried out, the beginning of the process and the rm In addition to the work carried out and the spoils were active.

En e u ca rio ta s , los te rm in o s "p ro m o to re s " o "p o te n c ia d o re s " se re fie ren a sen a les de con tro l de la tra n sc rip c io n . Los p ro m o to re s y p o te n c ia d o re s co n s is te n en m a trice s co rta s de se c u e n c ia s de A D N que in te ra cc io n a n e sp e c ifica m e n te con p ro te inas ce lu la re s im p lica da s en la tra n sc rip c io n (M a n ia tis et al., S c ience , 236 :1237 , 1987). S e han a is lado e le m e n to s p ro m o to re s y po te n c ia d o re s de una va rie d a d de fu e n te s euca rio tas , in c luye nd o genes de ce lu las de levadura , insecto , m a m ife ro y ve g e ta le s . T a m b ie n se han a is lad o e le m e n to s p ro m o to re s y p o te n c ia d o re s de v irus. En p ro ca rio tas ta m b ie n se e n cue n tran e lem e n tos de con tro l an a logo s , ta le s com o p ro m o to re s y p o tenc iad o res . La se lecc io n de un p ro m o to r y p o te n c ia d o r p a rticu la r de pende de l t ipo de ce lu la u tilizad o para e xp re sa r la p ro te ina de in te res. A lg u n o s p ro m o to re s y po te n c ia d o re s e u ca rio tas y p ro ca rio tas tien en una am p lia gam a de ce lu las h o sp e d a d o ra s de p ro du cc ion , m ie n tra s que o tros son fu n c io n a le s en un s u b co n ju n to lim itado de ce lu las h o sp e d a d o ra s de p ro du cc ion (vease, por ejemplo, V oss e t al., T re n d s B iochem . Sci., 11 :287 , 1986; Maniatis et al., 1987, a n te rio rm e n te c itado). Las e xp res ione s "e le m e n to p ro m oto r", "p rom o to re s ", o "se cu e n c ia p rom oto ra ", se re fie ren a una s e cu e n c ia de A D N que fu n c io n a co m o un in te rru p to r que a c tiva la e xp re s io n de un gen. Si el gen esta ac tivado , se d ice que se transc ribe , o que pa rtic ipa en la tra n sc rip c io n . La tra n sc rip c io n im p lica la s in tes is de A R N m a p a rtir de l gen. P o r lo tan to , el p rom oto r, s irve com o un e le m e n to re g u la d o r de la tra n sc r ip c io n y tam b ien p ro p o rc io n a un sitio para el in ic io de la tra n sc r ip c io n del gen en el A R N m .

In the United States, the te rm in os "p ro mo to re s" or "po te nc ia do re s" are referred to by the traffic control team. n. P ro mo to re sypo te nc ia do re s co ns te n in ma trice s corta of DNA se cts that in te c tio nane sp e nf e c tive nf e c tive with p ro te ins ce The implication is in the process (M an ia tis et al., S c ience, 236: 1237, 1987). If they have a side of the road to the potential of a variety of risks in the world, in which they have yeast genes, insect, mammal and go geta le s. They have also been to the men and women of the world. In this case, it is also possible to find lem tures with other participants, such as the tenders and tenants. The lecture of a p r mopo rypo te nc ia dorpa rticu la r de pende de lt ipo de ce lu u used for e xp re sa r p p te te in in res. Some people have a chance to meet them and they are in a friendly way with the houses that have been approved by the government, more than After all, they are not only in a limited sub-section of the housing services (see, for example, Vs et al., T re nds B iochem. Sci., 11: 287, 1986; Maniatis et al., 1987, before rm in te te cted). The e xp res ione s "e le men to p ro m oto r", "p rom o to re s", or "se cu enc ia p rom oto ra", refer to a se c on DNA that was necessary as an initiative that activates the gene expression. If the gene is activated, it is said that it is trans- ferred, or that it is part of the trait sc rip c io n. The tra c sc rip c io n imp p lica is the synthesis of RNA map rtir de l gen. For the most part, this is the place in which the site of the work is also available as well as a place to start io of the tra n sc r ip c io n of the gene in RNA m.

El te rm in o "p rop ied ad ", en el con tex to de una p ro te ina , se re fie re a cu a lq u ie r ca ra c te ris tica o a tr ib u to de un p o lin u c le o tid o que pued a se le c c io n a rs e o d e te c ta rse . E stas p ro p ie d a d e s in c luyen , pero sin lim itac ion , una p ro p ieda d que a fe c ta a la un ion a un po lip ep tido , una p ro p ieda d con fe rid a a una ce lu la que co m p re n d e un po lin uc leo tido pa rticu la r, una p rop iedad que a fe c ta a la tra n sc rip c io n de ge ne s (p o r e jem p lo , fu e rza de l p rom oto r, reco no c im ie n to de l p rom oto r, reg u lac io n de l p rom oto r, fu n c io n po tenc iad o ra ), una p rop iedad que a fe c ta al p ro ce sa m ie n to de A R N (p o r e jem p lo , corte y e m p a lm e de A R N , e s ta b ilida d de A R N , c o n fo rm a c io n de A R N , y m od ifica c ion po s tra n sc rip c io n a l), una p rop iedad que a fe c ta a la tra d u cc io n (p o r e jem p lo , n ivel, reg u lac io n , un ion de A R N m a p ro te in a s r ib oso m icas , m o d ifica c io n p o s tra du cc ion a l). P o r e jem p lo , un s itio de un ion pa ra un fa c to r de tra n sc rip c io n , una po lim e rasa , un fa c to r regu lado r, y s im ila res , de un p o lin uc leo tido , puede a lte ra rse para p ro d u c ir ca ra c te ris tica s d e se a d a s o para id e n tifica r ca ra c te ris tica s no deseab les .

The rm in or "p rop ied ad" tea, in the context of a p rst, will refer to what is the most important or tr tr u u r What can you be told? These are, but without limitation, a limitation that, at the time of the union, of an epitomized lip, a pledge of faith with a ridiculous faith. That is the reason that a small group has read it for a period of time, a company that at the end of the year has become the most important (for the first time, it is possible). of lp rom oto r, reco no c im ie np of lp rom oto r, region of lp rom oto r, fu nc io n po tenc iad (ra), a property that fe c ta For the sake of RNA (for example, for the cutting of RNA, it is also for RNA, with RNA purpose, and for the odors of this process). sc rip c io nal), a property that at the time of the work (for example, level, region, an RNA map map, as well as it was icas, modifies c io npos tra du cc ion al). P ore jem p lo, an ion site for a work fara t sc rip cio n, a small limo, a regular fa r to r, and im ila res, of a If you read it, you may want to go to the country to get rich in order to identify yourself as unwelcome.

El te rm in o "p ro p ie d a d ", en el co n te x to de una p ro te ina , se re fie re a cu a lq u ie r ca ra c te ris tica o a tr ib u to de una p ro te ina que pued a s e le cc io n a rse o de tec ta rse .

The r rm in o "p ro p e rity", in the context of a p r o te rn, refers to what is the most critical or tr One person who can tell you how to be born or of tech.

En el p re sen te docum e n to , los te rm in o s "p ro te in a " y "p o lip e p tid o " se u tilizan in d is tin tam en te . A lo la rgo de la p re sen te d ivu lg ac ion , para los res tos de am ino ac id os , se u tiliza el cod ig o de 3 le tras as i com o el cod ig o de 1 le tra co m o se de fin e de c o n fo rm id a d con la C o m is io n C o n ju n ta sob re N o m e n c la tu ra B io q u im ica (JC B N ) de la IU P A C -IU B . T a m b ie n se en tiende que, deb ido a la de ge ne ra c io n de l cod ig o gene tico , m as de una secu en c ia de p o lin uc leo tidos pued e un c o d ifica r un po lipep tido . U na e n z im a es una pro te ina .

In the document, the rm in o s "p ro te in a" and "p o lip e p tid o" are used in d is tin tam in te. In accordance with the law of the law, for the rest of the years, the code of 3 is used after that as well as the code of 1 How to end the con rm id ad with the C om is io n C on ju n ta sob re N omenc la tu b io quA ica ica (JC BN) of the UI PAC-IU B. It is also understood that, due to the generation of the genetic code, more than one section of the population can be coded for a few years. lipid tido. U na e n z im a is a protein.

En el p re sen te d o cum e n to , los te rm in o s "re n d im ie n to p ro p o rc io n a l" y "re n d im ie n to po rcen tu a l" se u tilizan in d is tin tam en te . S e re fie ren a la can tidad de un p roduc to de sea do en re lac io n con o tros p roduc tos que estan den tro de la m ism a m ezc la p ro du c ida po r un h o s p e d a d o r re co m b in a n te de la p resen te d ivu lg ac ion . P o r e jem p lo , el re n d im ie n to p ro p o rc io n a l de un p ro du c to de sea do puede m e jo ra rse de m odo que sea m as p re do m ina n te sob re los o tros c o m p o n e n te s en la m ezc la de l p ro du c to para re d u c ir la ca rg a de la pu rifica c ion . En o tro e jem p lo , el ren d im ien to p ro p o rc io n a l de un p ro du c to no de sea do (es decir, un com p o n e n te que d e be ra e lim in a rse de l p ro du c to de sea do ) puede re d u c irse de m od o que sea m en os p re do m ina n te sob re el co m p o n e n te de sea do en la m ezc la de l p roducto para lo g ra r el m ism o fin. En el p resen te d o cu m e n to los ren d im ien to s p ro p o rc io n a le s se exp resan en fo rm a de "X fre n te a o tros de riva d o s de ac idos g rasos", que com p ara n la can tidad de X, que es un tipo de de riva do de ac ido g ra so (p o r e jem p lo , un e s te r g raso , un d e riva d o de a c id o g ra so de una lo ng itud de ca d e n a pa rticu la r), y la exp res ion "o tro s de riva dos de ac idos g ra sos" s ig n ifica la can tidad a n ad ida de to d o s los de m as de riva dos de ac ido s grasos d is tin to s de X que se p ro du cen en el m ism o e xp e rim e n to , cu ltivo o c ic lo de fe rm en tac io n .

In the p re sen te do cum en to, the te rm in os "re nd im ie n to p ro po rc io nal" and "re nd im ie n to po rc tu al al" are used in d is tin also in tea It is true to the amount of a product of a person in control with other products that are within the same period of time as a host for co mb in an te de la p resen te d ivu lg ac ion. P o jem p lo, the re nd im ie n to p ro po rc io nal of a p ro du c t of sea do can be improved in a way that is more p re do m ina n te so re re the or other components in the department of the French Republic to reduce the status of the government. In other cases, the legal representation of a person's property (that is, a component that will be limited to the law) ro du c to de do do) it can be reduced from m od or that it is m in os p re do m ina n on the co m t of the do in the m ezc that of lp roducto for lo g ra r The same is the end. In the report, the ren- im rents of sp ro po rc io na s s are reviewed in the form of "X fre n te ao tros de riva dos de acid acids gos", which com For the amount of X, which is a type of source of interest (for example, one of these, a source of interest for one thing) ng itud de ca dena pa rticu la r), and the exp ion "o tro s de riva dos of acidic acids" means the amount added to all those of more riva two of the fatty acids of X which are carried out in the same period, in which quantum of faith in tac io n.

El te rm in o "p ro se q u e n c ia " se re fie re a una secu en c ia de am in o a c id o s e n tre la se cu e n c ia sena l y la p ro te ina m adura que es ne cesa ria para la sec re c io n de la p ro te ina . La esc is ion de la p ro se cu e n c ia puede c o n d u c ir a una p ro te in a /e n z im a a c tiva m adura en c ie rta s c ircu n s ta n c ia s y co n d ic io n e s ad ecu ad as .

The rm in o "p ro se que c ia" refers to a section in the middle of the in A A ³ n A A A A A A A A ± A A A A A A A ¡A A A ¡A ¡nA A nA A A A A A A A A A A A A A A A A A A A‚A‚AA‚AA‚A‚AA AM A? for the sec- tion of the church. The analysis of the project can be carried out for a short period of time in the center of the capital and also for the rest of the year.

El te rm in o "re co m b in a n te " cu a n d o se usa para m o d ifica r el te rm in o "ce lu la " o "ve c to r" en el p resen te docum e n to , se re fie re a una ce lu la o a un v e c to r que se ha m o d ifica d o in tro d u c ie n d o una s e cu e n c ia de p o lin u c le o tid o s he te ro loga , o que la ce lu la p rocede de una ce lu la que se ha m od ifica do de esta m anera . P o r tan to , po r e jem p lo , las ce lu las re co m b in a n te s exp resan ge ne s que no se en cu e n tra n en la m ism a fo rm a de n tro de la fo rm a na tiva (no re co m b in a n te ) de las ce lu las o, com o resu ltad o de una in te rve n c io n hu m an a in te nc iona da , exp resan genes na tivos que, de lo con tra rio , se exp resan de fo rm a anom a la , se in fra exp resan o no se exp resan en abso lu to . Los te rm in o s "re co m b in a c io n " "re co m b in a n d o " y la g e ne rac io n de un po lin u c le o tid o "re co m b in a d o ", se re fie ren g e n e ra lm e n te al e n sa m b la je de dos o m as fra g m e n to s de p o lin u c le o tid o s en dond e el e n sa m b la je da lu g a r a un po lin uc leo tido q u im e rico fa b rica d o a p a rtir de las pa rtes en sam b lad as .

The rm in or "re co mb in an te" tea is used to modify the rm in or "ce lu la" or "ve c to r" tea in the docum te pum in all, re faithful re to a city or a neighbor that has been modified in tro duce a se se c ony of l o l u l le le tid os ro lo lo lo, or that the ce la p roce of a It is the one that has been modified in this way. P o so to, po re jem p lo, the ce r re m m in an te s exp res ge ne s that i dont know how much in the same is fo rm a of the fo rm a national (not re co mb in an te) of the cities, com, or resu lt or of an inte r nn tio n h e an inte n tio n, there are native genes that, of otherwise, they expire from one to another, they enter or do not expire in abso luto. The rm in os "re co mb in ac" n "" re co mb in ando "and the generation of a small group" re co mb in ado ", re faith ren gene ra lm en te al in sa mb the two-man or the gmen fra s of po lin uc le o tid os in dond e el in sa mb the je da lu gara a po lin uc read tido qu im e rich fa b rich doapa rtir of the parts in sam b lad as.

Las e xp re s io n e s "se g m e n to reg u lado r", "se c u e n c ia s re g u la d o ra s ", o "se c u e n c ia de con tro l de e xp res ion ", se re fie ren a una se cu e n c ia de p o lin u c le o tid o s que es ta un ida o p e ra tiva m e n te a o tra s e cu e n c ia de p o lin u c le o tid o s que cod ifica la secu en c ia de a m in o a c id o s de una cad en a de po lip e p tid o s para e fe c tu a r la exp res ion de esa se cu e n c ia de a m in o a c id o s cod ifica da . La se cu e n c ia re g u la d o ra puede inh ib ir, rep rim ir, p rom ove r, o in c luso d ir ig ir la exp res ion de la se cu e n c ia de p o lin u c le o tid o s un ida o p e ra tiva m e n te que cod ifica la se cu e n c ia de am ino ac id os .

The e xp re s io nes "se gmen to reg u side r", "is counted s re gu la do ra s", or "is counted as con tr l of e xp res ion", re faithful re at one point it is possible that it is an operative step out of the way that codifies the sequence that codifies the sector in am o o The rights of a cad in a group of people for the faith of the expiration of that body are in accordance with the codified codes. The guideline can guide you to go, go to the country, go to the street, or even use the expiration of the signal to get it right now. an operative step that codifies the signal of love.

Las exp re s io n e s "m a rc a d o r de se le cc io n " o "m a rc a d o r se lec tivo ", se re fie ren a un p o lin uc leo tido (p o r e jem p lo , un ge n ) ca p a z de e xp re sa rse en una ce lu la h o spe da do ra , lo que pe rm ite fa c ilita r la se lecc io n de aq u e llo s h o sp e d a d o re s que con tien en el vec to r. C o m o e jem p los de m arca d o re s de se lecc io n se inc luyen , en tre o tros, m a rca do res a n tim ic ro b ia n o s . P o r tan to , la e xp re s io n "m a rc a d o r de se le cc io n " se re fie re a un gen que p ro p o rc io n a una in d ica c io n cu a n d o una ce lu la h o sp e d a d o ra ha ca p ta d o una se cu e n c ia e n tra n te de in te res o cu a n d o se ha p ro d u c id o a lgu na otra reacc ion . N o rm a lm e n te , los m a rca d o re s de se lecc io n son genes que con fie re n res is te nc ia a n tim ic ro b ia n a o una v e n ta ja m e ta b o lica en las ce lu las ho sp e d a d o ra s para p e rm itir que las ce lu la s que con tien en las secu en c ias e xo ge na s se d is tin g a n de las ce lu la s que no han rec ib id o las s e cu e n c ia s exogenas . Un "m a rc a d o r de se lecc ion res ide n te " es uno que se lo ca liza en el c ro m o so m a de l m ic ro o rg a n ism o que se va a trans fo rm a r. Un m a rca d o r de se lecc io n res iden te co d ifica un gen que es d ife re n te de l m a rc a d o r de se le cc io n en la con s tru cc io n tra n s fo rm a d o ra . Los e xp e rto s en la m a te ria conocen m a rca d o re s se lec tivo s . C o m o se in d ico an te rio rm e n te , a d e cu a d a m e n te el m a rc a d o r es un m a rc a d o r res is te n te a a n tim ic ro b ia n o s , in c luye nd o , po r e jem p lo , a m p R; f le o R; e s p e c R; k a n R; e r iR; te tR; c m p R; y n e oR. V ea se , po r e jem p lo , G u e ro t-F le u ry , G ene, 167 :335 -337 , 1995; P a lm ero s e t al., G ene, 247 :255 -264, 2000 ; y T r ie u -C u o t e t al., G ene, 23 :331 -341 , 1983. O tro s m a rca d o re s u tiles de acu e rd o con la d ivu lg ac ion inc luyen, pero s in lim itac ion , m a rca d o re s a u xo tro fos , ta le s com o trip to fa n o ; y m a rca d o re s de d e tecc io n , ta le s com o 6-g a la c to s ida sa .

The exp "s" ma rc ador de se le cc io n "or" ma rc ador se se le t tive ", refer to a small group of people (for example, a person) peace From this point of view, in a city, the home of what happened is the lesson of which the people who have been in the neighborhood. C o m o e jem p those of the arca d o re s se lection are included, among others, m a rca do res a n tim ic ro b ia n o s. For example, the e xp re s io n "ma rc ador de se le c io n" refers to a gene that has only an idyllic idea when a ce A resident has been able to sign up in the process of interest or when there has been another reaction. N o rm a lm en te, the macaca re re s se nion are genes that, with faith re n res is te nc ia an tim ic ro b ia nao, a ve ve ta ja me ta basica in the ce They have been hoped to see that they are the ones they have in the sectors that have been granted the decisions that they have not received. It is exogenous. A "m a rc a d o r de se lection res ide n te" is one that can be used in the same way as the one that is going to be transferred. A brand of identifiable languages is a gene that is different from the one in which it is read in the con- structor of the gene. The e xp e rts in the m a ria know m a rca d o re s se le t tive s. How rm in te rio in te was introduced, in addition to the fact that the master is a master who is important to us, in which he or she is not able to do so, , amp R; f le or R; e s p e c R; k to n R; e r iR; te tR; c m p R; and n e oR. See, po r e jem p lo, G u e ro t-F le u ry, Jan, 167: 335-337, 1995; P a lm ero s e t al., Jan 247: 255-264, 2000; and T r ie u -C uotet al., G Jan, 23: 331-341, 1983. Another reason for the use of water is the same, but without lim ITAC ION, MARCA DE RE SAU XO TRO FOS, TER S S COM OR TRIP TRO FA NO; and more than 6 months, the com 6a to the first leg.

La exp res ion "se cu e n c ia de nu c le o tido s que cod ifica un m a rc a d o r de se le cc io n " se re fie re a una secu en c ia de p o lin u c le o tid o s que es cap az de e xp resa rse en las ce lu las ho sp e d a d o ra s y donde la exp res ion del m a rc a d o r de se lecc io n co n fie re a las ce lu las que con tienen el gen exp resa do , la cap ac idad de c re c e r en p re sen c ia de un agen te se le c tivo co rre sp o n d ie n te o en a u se n c ia de uno o m as nu trien te s esenc ia les .

The expression "is made up of the nuances encoding a master of the seer" refers to a section of the population that is The captain of the xp resa rse in the houses of the pastors and where the expiration of the master of the lesson with those who have the expired gene, the CAPACITY OF REPENSION IN A REPRESENTATIVE OF AN AGENT IS LEARNED IN SPIRITUAL OR IN ORDER OF ANOTHER OUR NUMBER THREE ESSENTIALS.

U na "se cu e n c ia sena l" o "p ep tido sen a l" se re fie re a una se cu e n c ia de po lin u c le o tid o s o de am in o a c id o s que pa rtic ip a en la secre c io n de las fo rm a s m a d u ra o p re cu rso ra de una p ro te ina . Esta de fin ic ion de se cu e n c ia sen a l es fun c ion a l, y s ig n ifica que in c luye to d a s aq u e lla s se cu e n c ia s de a m in o a c id o s c o d ifica d a s po r la pa rte N -te rm ina l del gen de la p ro te ina , que pa rtic ip an en la rea liza c io n de la sec re c io n de p ro te inas . A m enudo , pero no g lob a lm e n te , es ta n u n id as a la pa rte N -te rm in a l de una p ro te in a o a la pa rte N -te rm in a l de una p ro te ina p re cu rso ra . La se cu e n c ia sen a l puede s e r e n d o ge na o exogena . La se cu e n c ia sen a l puede s e r una que no rm a lm e n te esta aso c ia d a con la p ro te ina (p o r e jem p lo , t io e s te ra sa ), o puede s e r una o rig in a d a o que p rocede de un gen que co d ifica o tra p ro te ina sec re ta da . Una secu en c ia sena l exo ge na e je m p la r com p ren de los s ie te p rim e ros res tos de a m in o a c id o s de la se cu e n c ia sena l de sub tilis in a de Bacillus subtilis fu s io n a d a con el resto de la secu en c ia sena l de la sub tilis ina de Bacillus lentus (A T C C 21536). O tra s e cu e n c ia sen a l e je m p la r c o m p re n d e la s e cu e n c ia sena l de T e s A que se e lim in a para p ro d u c ir 'TesA .

A "be aware of the signal" or "pertaining to the referent" refers to a signal from the United States or those that the IP address the secretariat of the asmadura op op cu qura ra of a p ro te ina. The end of this statement is that it is func- tional, and it means that it includes all of those who are involved in the process of decoding them. For the N-te rm ina l of the gene of p ro te ina, which pa rtic ip an in the realm of the section of p ro te inas. At the same time, but I don't know how to do it, it is still a part of the N-te rm in al of a p ro te in aoa the N-te rm in al of a p ro te ina p re cu rso ra. The signal can be learned or exogenous. The signal may be one that does not rm a lm in which you are associated with the group (for example, this is the case), or it may be one or more rigid. that comes from a gene that encodes or works for you. A section in the signal exo ge na e je mp the r com p r of the se s p te r p e r e s ress of am in or c of the signal of the signal of sub tilis in a of Bacillus subtilis was nothing with the rest of the secu in the sign of the sub tilis of Bacillus lentus (ATCC 21536). Another issue is that the rcomp re nde the signal with the sign of T is A that is limited for the purpose of going to TesA.

La exp res ion "su s ta n c ia lm e n te id en ticos", en el con tex to de dos po lin u c le o tid o s o dos po lip ep tido s , se re fie re a un po lin u c le o tid o o a un p o lip ep tido que com p ren de una id en tidad de secu en c ia de al m en os 70 %, po r e jem p lo , al m en os 75 %, al m en os 80 %, al m en os 85 %, al m en os 90 %, al m enos 91 %, al m en os 92 %, al m en os 93 %, al m en os 94 %, al m en os 95 %, al m en os 96 %, al m en os 97 %, al m en os 98 % o al m en os 99 % de id en tidad de se cu e n c ia en c o m p a ra c io n con una se cu e n c ia de re fe re nc ia u tiliza n d o los p ro g ra m as o a lg o ritm o s (p o r e jem p lo , B LA S T, A L IG N , C L U S T A L ) u tilizan do p a ram e tro s es tanda r. Una in d ica c io n de que dos po lip ep tido s son su s ta n c ia lm e n te id e n tico s puede s e r que el p r im e r p o lip ep tido te n g a in m u n o rre a c tiv id a d c ru za d a con el seg un do po lip ep tido . N o rm a lm e n te , los po lip ep tido s que d ifie ren po r su s titu c io n e s de a m ino ac id os con se rva tivas , tien en in m u n o rre a c tiv id a d c ruzada . P o r tan to , un p o lip e p tid o es s u s ta n c ia lm e n te iden tico a un seg un do po lipep tido , por e jem p lo , cuando los dos pe p tid os d ifie ren so lo en una su s titu c io n con se rva tiva . O tra in d ica c io n de que dos s e cu e n c ia s de p o lin u c le o tid o s son su s ta n c ia lm e n te iden ticas es que las dos m o lecu las se h ib rid an en tre s i en co n d ic io n e s rig u rosa s (p o r e jem p lo , en un in te rva lo de r ig u ros ida d m ed io a m axim o).

The expression "your s ta nc ia lm in identical te", in the context of two po u u le le tid bear two po lip ep tido s, refers to a po lin uc le o t o o a po lip epid t that has a seity identity of at least 70%, at least 75%, at 80%, at 80%, at m in os 85%, to m in os 90%, to m inos 91%, to m in os 92%, to m in os 93%, to m in os 94%, to m in os 95%, to m in os 96%, to m in os 97%, to m in os 98 % or to m in os 99 % of the identity of the person in charge Comparison with a re c n re c cio n re c n u ia or use of the p ro g ra m as or lg or rhythms (for example, B LA ST, AL IG N, CLUSTAL) or use do pa ram e tro s is tanda r. One indication that two epithelial lipids are their only importance may be that the epiphytic lipprince is denied to you in many cases. he gives with the second one epid lipid. N o rm a lm in te, the epiphytic poisons that d ifie ren ren r r its subtitle of love in se riv tives, have in rune ac tiv id adc ruc. Therefore, a po lip e p tid o is its ta lc lia lm in the same way to a seg a dopo lipo lipid, for example, when the two losses of ifie ren only so A substitution with se rva tiva. Another trait of the fact that the two are aware of their potential is that the two are the only ones that were ridiculous between them. In co nd ic io nes rig u rosa s (pore jem p lo, in a rva u r r u u ida dm ed io am axim o).

M o lecu las "su s ta n c ia lm e n te p u rifica das" s ig n ifica m o lecu las que al m en os a p ro x im a d a m e n te en un 60 %, p re fe re n te m e n te al m en os a p ro x im a d a m e n te en un 75 %, a p ro x im a d a m e n te en un 80 %, a p ro x im a d a m e n te en un 85 % y m as p re fe re n te m e n te al m en os a p ro x im a d a m e n te en un 90 %, carecen de o tros c o m p o n e n te s con los que es ta n a s o c ia d o s de fo rm a na tu ra l. T a l co m o se usa en el p re se n te do cu m e n to , los te rm in o s "p u rif ic a d o " o "pu rifica r", ta m b ie n se re fie ren a la e lim in a c io n de co n ta m in a n te s de una m uestra . P o r e jem p lo , la e lim in a c io n de co n ta m in a n te s pued e p ro d u c ir un a u m e n to en el p o rce n ta je de d e riva d o s de a c id o s g ra so s de in te res en una m uestra . P o r e jem p lo , de sp u e s de que los de riva dos de ac idos g ra sos se exp resen en ce lu la s ho sp e d a d o ra s de p ro du cc ion de p lan tas, bac te rias , le va d u ra s o m am ife ros , los d e riva d o s de ac ido s g ra sos pueden p u rifica rse , po r e jem p lo , m ed ian te e lim in a c io n de p ro te inas de la ce lu la h o spe da do ra de p ro du cc ion . Esta e tapa , d e n o m in a d a ta m b ie n e tapa de recu pe rac io n , im p lica la se p a ra c io n y el p ro ce sa m ie n to de la com p o s ic io n de d e riva d o s de ac ido s g ra sos de m odo que la com p o s ic io n sea util en a p lica c io n e s in du s tria le s , po r e jem p lo , com o co m b u s tib le o p ro du c to qu im ico . D e spu es de la pu rifica c ion , el po rcen ta je de d e riva d o s de ac ido s g ra sos en la m ue s tra se aum en ta . El te rm in o p u rif ica d o no requ ie re pu reza ab so lu ta ; m as bien, se en tiende com o un te rm in o re la tivo . P o r tan to , po r e jem p lo , una p re pa rac io n de de riva do de ac ido g raso p u rifica do es una en la que el p ro du c to esta m as co n ce n tra d o que el p ro du c to en su e n to rno de n tro de una ce lu la . P o r e jem p lo , un e s te r g ra so pu rifica do es uno que es ta s u s ta n c ia lm e n te se p a ra d o de c o m p o n e n te s ce lu la re s (p o r e jem p lo , p o lin u c le o tid o s , lip idos, h id ra to s de c a rb o n o y o tros p e p tid o s ) que pueden aco m p a n a rlo . En o tro e jem p lo , una p re pa rac io n de e s te r g raso pu rifica do es una en la que el e s te r graso ca re ce su s ta n c ia lm e n te de c o n ta m in a n te s , co m o los que puede h a b e r de sp u e s de la fe rm e n ta c io n . P o r e jem p lo , se d ice que un e s te r g ra so esta "p u rif ica d o " cu a n d o al m en os a p ro x im a d a m e n te el 50 % en peso de una m u e s tra esta c o m p u e s to po r el e s te r graso . En o tro e jem p lo , cua nd o al m en os a p ro x im a d a m e n te el 60 %, 70 %, 80 %, 85 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 96 %, 97 %, 98 % o el 99 % o m as en peso de una m u e s tra esta com p ue s to po r el e s te r graso.

M o lecu the "su s ta nc ia lm en te p rifica das" means only the ones that at the same time in 60%, for the most important time at the same time in ap ro x im adamen te in 75%, ap ro x im adamen te in 80%, ap ro x im adamen te in 85% and more p re faith re n te ment to me in ap ap ro x im adamen te in 90%, they lack or other components with which it is ta nas nas ia dos de fo rm a na tu ra l. As it is used in the past, the te rm in os "pu rif ic ado" or "pu rifica r", also refers to the e lim in ac Co n ta m in an te s of a sample. P o jem p lo, the e lim in ac tion of co n ta m in an te s can go ep ro duc go an increase in the power of riva n ta je de riva dos de a c id osg ra so s de in laugh at a sample. P o jem p lo, on the basis that those of riva dos de g acid are present at the same time as p o du p e r ents of p ro du cc ion du p lan tas, bac te rias , it is going to be easy for you, those from Riva dos de Ac A³ Sg Ra SOS can be certified, by the way they are also limited to the action of the churches. the hospitality of p ro du cc ion. This cover, called in addition to the cover of recovery, implies the separation and the interest of the community of de riva dos de de It is a matter of course that the com- position be useful in the practice of the trusts, as well as how to conduct the optic duo to the chemical. . In the case of the public, the power of human rights is also increased in the sample. The tea rm in or p rif ica d o does not require re pe reza ab so lu ta; rather, it is understood as a rm in or re tive tea. P o so to, po re jem p lo, a p re pa ration of public de riva do p o r is a one in which the property is more common that the p du du t in its environment within one of a city. P o jem p lo, an es rg ra so pu rifica is one that is your ta nc ia lm in which you are made up of the components (pore jem p lo, po lin uc le o tid os, lipids, h id ra to s of ca rb onoyo tros p e p tid os) which can accommodate it. In other cases, a p r pa r ration of this public property is one in which the fatty acid falls into its own state in relation to the con in an te s, like those that can be of sp u e s of the faith rm in ta c io n. P o jem p lo, it was said that one of these is "pu rif ica do" when he goes to me in about 50% by weight of a sample is composed of He is fat. In other cases, at least in 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95 %, 96%, 97%, 98% or 99% or more by weight of a sample is composed of fatty oil.

"S u s titu c io n " s ig n ifica re e m p la z a r un a m in o a c id o en la se cu e n c ia de una p ro te ina p re cu rso ra con o tro a m in o a c id o en una pos ic ion pa rticu la r, da n d o com o resu ltad o un m u tan te de la p ro te ina p re cu rso ra . El am in o a c id o u tilizado com o su s titu to puede se r un a m in o a c id o na tura l, o pued e s e r un a m in o a c id o s in te tico o no na tura l.

"S us titu c io n" s ig n ificare re emp la zar an am in o a c id o in the se c on of a p ro te na p re qu rso ra con o tro am in o a c id or in a position to pay for the damage, or damage to a person or a member of the family. The am in o a c id or used as its substitute can be a m a n in o c id o na tura l, or it can be a m a m in o a c in o a t in t in tico or no na tura l.

El te rm in o "te n s io a c tivo s " se re fie re a su s ta n c ia s que son ca p a ce s de re d u c ir la te n s io n su p e rfic ia l de un liqu id o en do nde se d isue lven . N o rm a lm e n te , se co m p o n e n de una cab eza s o lu b le en ag ua y de una cadena o co la de h id ro ca rbu ro . La cabeza so lub le en agua es h id ro fila y puede s e r ion ica o no ion ica . La cad en a de h id ro ca rbu ro es h id ro foba . Los te n s io a c tivo s se u tilizan en una va rie d a d de p roduc tos , in c luye nd o de te rg e n te s y lim p ia do res , y ta m b ie n se u tilizan com o a u x ilia re s para tex tile s , cue ro y pape l, en p ro ceso s qu im icos , en p ro du c to s co sm e tico s y fa rm a ce u tico s , en la in du s tria a lim en ta ria , en la ag ricu ltu ra , y en la recu pe rac io n de ace ites . A de m as , pueden u tiliza rse para a yu d a r en la ex tracc ion y el a is la m ie n to de ace ites c rud os que se e n cue n tran en en to rno s de d ific il acce so o en e m u ls io n e s de agua. H ay cua tro tip o s de te n s io a c tiv o s c a ra c te riza d o s po r d ife ren te s usos. Los te n s io a c tiv o s an io n ico s tien en una ac tiv idad s im ila r al de te rge n te y se u tilizan ge n e ra lm e n te para ap lica c io n e s de lim p ieza . Los te n s io a c tiv o s ca tio n icos con tien en h id ro ca rb u ro s de cad en a la rga y a m en ud o se u tilizan para tra ta r p ro te in a s y p o lim e ro s s in te tico s o son c o m p o n e n te s de s u a v iz a n te s pa ra te jid o s y a co n d ic io n a d o re s pa ra el cabe llo . Los te n s io a c tivo s an fo te ro s ta m b ie n con tien en h id ro ca rb u ro s de cad en a la rga y no rm a lm e n te u tilizan en cham pus. Los te n s io a c tiv o s no io n ico s a m en ud o se u tilizan en p ro du c to s de lim pieza.

The tea rm in o "te ns io ac tivo s" refers to its s ta nc ia s which are capable of reducing the cost of a liquid id in Where is it? N o rm a lm e n te, it is understood as a single head and a chain or with a chain of water. The head is only in water and can be ionic or non ionic. The body of h id ro ca rbu ro is h id ro foba. The active agents are used in a variety of products, in c te r e nd o te rg in te s and lim pia do res, and are also used as a product. for tex tile s, cue and paper l, for chemical reasons, for cooperative and professional purposes, in the industrial sector in Lima He laughed, in the agoutu lutra, and in the collection of acets. In addition, they can be used to help in the ex- traction and the year is the most important way to find them in the day of access. or in emu ls io nes of water. There are four types of te s s io a c tiv o s c a ra c te riza d o s po r d ife ren te s uses. Te n anio n c a s tio n s an annico s s s have a similar activity and are used for general purposes of cleaning. Ca tio n tio ns ca tio nicos with ten in h id ro ca rb u ro s de cad en a rga yam en u o are used to treat r t ro te in asypo limm e ro ss in tetico So they are composed of soften iz an te s pa ra te jid osya co nd ic io nado re s para la cabe llo. The te n s io a c tivo s an fo te ro s ta m b ie with ten a d ca rb u ro s of cad in a rga and not rm a lm e n use u in cham pus. The te n s io a c tiv o s are not unique to you or are used as part of your piece.

El te rm in o "s in ta sa " se re fie re a una en z im a que ca ta liza un p ro ceso de s in tes is . T a l com o se usa en el p resen te do cum e n to , el te rm in o "s in ta sa " in c luye s in ta sa s y s in te tasas .

The rm in or "s in ta sa" tea refers to one in which a period of syndromes is possible. As used in the present, the rm in tea or "s in ta sa" in c luye s in ta sa s and s in te rates.

La exp res ion "p ro p ie d a d d ian a " se re fie re a una p ro p ieda d del gen de pa rtida que se p re te n d e a lte rar.

The expression "p ro p ie d a d d ian a" refers to a p r p p eda d of the partid gene that is present in the rarity.

El te rm in o "tio e s te ra sa " se re fie re a una en z im a que tien e ac tiv id a d tio e s te ra sa . Las tio e s te ra s a s inc luyen t io e s te r h id ro lasa s , que se id e n tifica n co m o m ie m b ro s de la C la s ifica c io n de E nz im as , EC 3.1.2 , y que p u ed en o b te n e rs e de una v a rie d a d de fuen tes . P o r e jem p lo , en V o e lk e r y D av ies, J. Bact., Vol., 176, n.° 23, pags. 7320 -27 , 1994 y en las pa ten tes de E s tad os U n idos n.° 5.667.997 y n.° 5.455.167 , se de sc rib en t io e s te ra sa s ve g e ta le s . Las t io e s te ra sa s ta m b ie n pu ed en o b te n e rse de fu e n te s m ic ro b ia na s , ta le s co m o las d e sc rita s en A ko h e t al., P rog. L ip id Res., vo l. 43, n.° 6, pags. 534 -52 , 2004 ; D icz fa lu sy y A le xso n , A rch . B iochem . B iophys., vo l. 334, n.° 1, pags. 104-12, 1996; La rson y K o la ttukudy , A rch . B iochem . B iophys., vo l. 237, n.° 1, pags. 27 -37 , 1985; Law son et al., B io che m is try , vo l. 33, n.° 32, pags. 9382 -88 , 1994; Lee e t al., Eur. J. B iochem ., vo l. 184, n.° 1, pags. 21 -28 , 1989; N a gg e rt e t al., J. Biol. C hem ., vo l. 266, n.° 17, pags. 11044-50 , 1991; N ie e t al., B io che m is try , vo l. 47, n.° 29, pags. 7744 -51 ,2008 ; S ea y y Luek ing , B io che m is try , vo l. 25, n.° 9, pags. 2480 -85 , 1986; S p e n ce r e t al., J. Biol. C hem ., vo l. 253, n.° 17, pags.

The rm in tea or "uncle es te ra sa" refers to one in which you have ac tiv id ad unio es te ra sa. The uncle is te ra sas inc luyen un io e s te r r id id las las s s, which is identified in the form of m i m m ro s of the C the meaning of E nz im as, EC 3.1.2 , and that I am able to obtain a variety of sources. P ore jem p lo, in V oe lk ery D av ies, J. Bact., Vol., 176, no. 23, p. 7320-27, 1994 and in the patents of the United States No. 5,667,997 and No. 5,455,167, it is discussed in this context. . The guys are te sa s ta mb ie n pu ed in ob te ne de fu en te sm ic ro b ia na s, as well as those of sc rita s in A ko het al., P rog . L ip id Res., Vo l. 43, No. 6, p. 534-52, 2004; D icz fa lu sy and A le xso n, A rch. B iochem. B iophys., Vo l. 334, No. 1, p. 104-12, 1996; The rson and K or the ttukudy, A rch. B iochem. B iophys., Vo l. 237, No. 1, p. 27-37, 1985; Law son et al., B io che m is try, vo l. 33, No. 32, p. 9382-88, 1994; Lee et al., Eur. J. B iochem., Vo l. 184, # 1, p. 21-28, 1989; N a gg e rt et al., J. Biol. C hem., Vo l. 266, No. 17, p. 11044-50, 1991; N ie et al., B io che m is try, vo l. 47, no. 29, p. 7744-51, 2008; S ea yy Luek ing, B io che m is try, vo l. 25, No. 9, p. 2480-85, 1986; S pen ce ret al., J. Biol. C hem., Vo l. 253, No. 17, p.

5922 -26 , 1978; y en Z h u a n g et al., B io che m is try , vol. 47, n.° 9, pags. 2789 -96 , 2008. T am b ien pueden ob tene rse t io e s te ra sa s , po r e jem p lo , de c ia n o b a c te ria s , a lgas, m am ife ros , in sec tos y hongos. Una tio e s te ra sa puede te n e r ac tiv ida d d is tin ta de ac tiv idad tio e s te ra sa , po r e jem p lo , ac tiv idad p ro te o litica o ac tiv ida d de h id ro lis is de e s te r y ox igeno . Una tio e s te ra sa pa rticu la rm e n te util es la en z im a 'T e sA (o t io e s te ra sa I) de E. coli, que es una ve rs ion tru n c a d a de la e n z im a T e s A se rin a t io e s te ra s a T e s A de lo ng itud c o m p le ta que se de sc rib e en C ho y C ronan , J. B io l. C hem ., vo l., 268, n.° 13, pags. 9238 -45 , 1992. Un p o lip ep tido 'T e sA de E. coli c o m p re n d e 182 a m in o a c id o s , y es el p ro d u c to de una re a cc io n de e sc is io n en d o n d e se e lim in a la s e c u e n c ia lid e r de 26 a m in o a c id o s de T e s A de E. coli. El p o lip e p tid o 'T e s A de E. coli, po r e jem p lo , t ie n e la s e cu e n c ia de a m in o a c id o s de la S E Q ID NO: 31 e xp u e s ta en la FIG. 58.

5922-26, 1978; and in Z huang et al., B io che m is try, vol. 47, No. 9, p. 2789-96, 2008. You can also obtain uncle es te ra sa s, po re jem p lo, from c a nobac te la s, to lgas, m am ife ros, in sec coughs and fungi. An uncle is te ra sa can have ac tiv ida dd is tin ta de ac tiv idad un un es ra ra, po re jem p lo, ac tiv idad p ro te o litica o ac tiv ida d de h id ro lis It is from you and oxygen. An uncle is te r sa pa rticu the rm in te useful is the en z im a 'T e sA (ot io es te ra sa I) of E. coli, which is a tru nion of the enzyme im T es A surrender at t es te ra sa T es A from the ng itud comp le ta that is sc rib e in C ho and C ronan, J. B io l. C hem., Vo l., 268, no. 13, p. 9238-45, 1992. A small group of E coli T e sA from 182 am in o c ids, and is the product of a re e ctio n of e sc isio n where it is limited to the 26 am in line sequence or T of A is E. coli. The po lip e p tid o 'T is A from E. coli, po re jem p lo, t ie ne the se am o of am in o c o id of SEQ ID NO: 31 e xp uesta in FIG. 58.

La exp res ion "a c tiv id a d tio e s te ra s a " se re fie re a la c a p a c id a d de c a ta liz a r una rea cc io n de esc is io n de l t io e s te r, que g e n e ra lm e n te im p lica la h id ro lis is de un t io e s te r en un g ru p o tio l en un ac ido y un tio l, pero ta m b ie n pued e in c lu ir una e ta pa de tra n se s te r if ica c io n en la que se esc inde un en la ce t io e s te r y se fo rm a un nuevo en lace este r. En genera l, una a c il-A C P tio e s te ra s a es cap az de c a ta liz a r la esc is ion h id ro litica de tio e s te re s de ac il p ro te in a tra n s p o rta d o ra de ac ilo y /o tio e s te re s de a c il-co e n z im a A grasa . C o m o e jem p los de e n z im a s que tie n e n ac tiv ida d t io e s te ra s a se in c luye n a ce til-C o A h id ro lasa , p a lm ito il-C o A h id ro lasa , su cc in il-C o A h id ro lasa , fo rm il-C o A h id ro lasa , ac il-C o A h id ro lasa , pa lm ito il-p ro te in a tio e s te ra s a y ub iq u itin a tio le s te ra sa . La ac tiv ida d t io e s te ra sa puede es ta b le ce rse m ed ia n te cu a lq u ie ra de los s ig u ie n te s ensayos:

The term "ac tiv id ad un un es es ra ra" refers to the capacity of the ca l l l e l a rea sion reaction of lt io e s te r, which generates lm in imp lica the h id ro lis is of an un io e s te r in a g ru po unio l in an acid and an uncle l, but ta mb ie n can go in c lu go a e ta In this sense, it is the case in which a discussion is made in the ce r io e s te r r and a new one is established in the latter. In general, an ac-cio CP unio es te ra sa is the capital of the political analysis of uncle e s te re s of ac il p ro te in through the nspo rta do Acyl and / or uncle is a fatty acid. Like jem p those of enzymes that have ac tiv ida dt io es te ra sa se in c luye na ce til-C o A h id ro lasa, pa lm ito il-C or A h id ro lasa, your cc in il-C or A h id ro lasa, fo rm il-C or A h id ro lasa, ac il-C or A h id ro lasa, pa lm ito il-p ro te in a uncle es te ra say ub iq u itin a uncle le s te ra sa. The activity of this process may also be carried out by the essays of the following essays:

Ensayo de hidrolisis de acil-CoA:

Acyl-CoA hydrolysis test:

P ara p re p a ra r una m ezc la de en sayo com p le ta , se u tiliza un ta m p o n T ris -H C l, 0,1 M, pH 8,0; P a lm ito il-C o A , 5 pM; D TN B , 0,01 M en ta m p o n de fo s fa to de po tas io 0,1 M, pH 7,0. P o r tan to , la m ezc la de e n sayo con tien e una c o n ce n tra c io n fina l de 10 p m o l de ta m p o n T ris -H C l, pH 8,0, 0 ,05 pm o l de D TN B y 0,01 pm o l de pa lm ito il-C o A . D espues, la m ezc la de e n sayo c o m p le ta se m ezc la con la tio e s te ra sa , en un v o lu m e n fina l de 2 ,0 ml.

For a comparison of a co-test, a T ris-H C l, 0.1 M, pH 8.0 is used; P a lm ito il-C or A, 5 pM; D TN B, 0.01 M at the same time of 0.1 M, pH 7.0. For example, the test in question has a fine concentration of 10 pmol of T ris-HC l, pH 8.0, 0.05 p.m. D TN B and 0.01 pm ol of pa lm ito il-C or A. Then, the m ezc that of e n sayo c o m p le ta m ezc la with uncle e s te ra sa, in a fine time l of 2.0 ml.

La v e lo c id a d de e sc is io n de l sus tra to a c il-C o A se m ide co n tro la n d o el c a m b io en la a b so rb a n c ia a 405 nm, u tilizan do un co e fic ie n te de ex tin c io n m o la r de 13 600 M-1 cm -1.

The view of the sciences of his C-C or A treatment is shown as the change in the 405 nm abb ance, using a co In fact, it is the r of 13 600 M-1 cm -1.

Ensayo in v iv o :

In v iv test or:

La tio e s te ra sa de in te res se exp resa en un h o sp e d a d o r adecu ad o , ta l com o E. coli. D e spu es de la exp res ion de la p ro te ina , el cu ltivo se ac id ifica con HCl 1 N hasta un pH fina l de a p ro x im a d a m e n te 2 ,5 y de spu es se ex trae con un m ism o vo lu m e n de a ce ta to de etilo . Los ac ido s g ra sos lib res en la fase o rg a n ica se d e riva tizan con h id ro x ido de te tra m e tila m o n io (T M A H ) para g e n e ra r los es te re s m e tilico s resp ec tivo s , que de sp u e s se ana lizan en un c ro m a to g ra fo de ga ses d o tado de un d e te c to r de io n iza c io n de llam a.

Uncle es te ra sa de in te res is expressed in an appropriate age, such as E. coli. After the expiration of the project, the quantification is validated with 1 N HCl up to a fine pH of approximately 2 and 5 and then it is extracted with a very low volume of ethyl acetate. Sg ra are free in the phase or rg a n ica riva tizan with h id ro x gone te te me me tila mon io (TMAH) to generate the re te s t he s til t s s e s ec Yes, they are analyzed in a major way of guaranteeing a call center.

Ensayo de hidrolisis de tiolactona:

Thiolactone hydrolysis assay:

En p r im e r lugar, se p re pa ra una so luc io n de rea c tivo que co n tie n e L -h o m o c is te in a t io la c to n a (L -H cyT ) 25 m M y ac ido 5 ,5 -d itio -b is -2 -n itro b e n z o ic o (D T N B ) 0 ,5 m M en ta m p o n H E P E S 0,1 M (pH 7,3). D espues, a la so luc io n de reactivo se la anade en z im a y la h id ro lis is de L -H cyT se con tro la d e te c ta n d o el g rupo tio l lib re con D TN B a 412 nm ( = 13 600 M-1 cm-1 pa ra el ac ido 5 -tio -2 -n itro b e n zo ico ).

First of all, there is a reactive solution that takes place in the city (L-H cyT) 25 mM and ac 5 , 5-d itio -b is -2 -n itro benzo ic or (DTNB) 0.5 mM in size 0.1 M HEPES (pH 7.3). Then, to the reagent solution, it is added in zim a and the r h lols of L-H cyT is shown in the same way as the group of people with D TN B a 412 nm ( = 13 600 M-1 cm-1 for the acid 5 -tio -2 -n itro ben zo ico).

Ensayo de 4-MU-6S-Palm-pGlc:

4-MU-6S-Palm-pGlc test:

En p r im e r lugar, se p re pa ra una m ezc la de rea cc io n que co n tie n e 10 pl de e n z im a tio e s te ra s a y 20 pl de so lu c io n de sus tra to . La so luc io n de su s tra to con tien e M U -6 S -P a lm -p -G lc 0 ,64 m M , d itio tre ito l (D T T ) 15 m M , T riton X -100 al 0 ,375 % (p /v ) y 0,1 U de p -g lu co s id a sa de a lm e n d ra s en tam po n fo s fa to /c itra to de M c llva in , pH 4,0. La m ezc la de rea cc io n se in cuba d u ra n te 1 hora a 37 °C. S e anad e p -g lu co s id a sa de a lm e n d ra exo g e n a para h id ro liza r el p ro du c to in te rm e d io de reacc ion , M U -6 -tio -p -g lu co s id o , cua n tita tiva m en te . La reacc ion de h id ro lis is fina liza con la ad ic ion de 200 pl de ca rb o n a to de sod io 0 ,5 M, pH 10,7, que con tiene T rito n X -100 al 0 ,025 % y la f lu o re sce n c ia de la 4 -m e tilu m b e life ro n a (M U ) libe ra da se m ide en un flu o ro m e tro (Aex = 372, Aem 445 nm).

In the first place, there is a reaction that takes place in 10 years of the work and 20 pl of their work. The solution of its system with MU -6 S -P a lm -p -G lc 0.64 mM, di thio tre ito l (DTT) 15 mM, T riton X -100 al 0, 375% (w / v) and 0.1 U of p-g lu co s id a sa from a lm end ra s in m a s p to fa c / m itra to M c llva in, pH 4 , 0. The reaction mixture is incubated for 1 hour at 37 ° C. Add ep -g lu co s id a sa de a lm end ra exo gena for h id ro liza r el p ro du c to in te rm ed io reaction, MU -6 -thio -p -g lu How do you know, how much you are. The reaction of human idols is fine with the addition of 200 µl of 0.5M sodium carbonate, pH 10.7, which with Rite n X -100 at 0.025 % and the f lu o re sce nc ia of the 4-me tilu mbe life ro na (MU) is released in a flow or meter (Aex = 372, Aem 445 nm).

Ensayo de lisofosfolipasa:

Lysophospholipase Assay:

S e p repa ra una m ezc la de reacc ion que con tiene 10 pl de t io e s te ra sa m ezc lad a con 10 pl de 1 -o leo ilfo s fa tid ile ta n o la m in a 3 m M , 25 pl de T ris -H C l 100 m M (pH 7 ,0 ) y 5 pl de E D T A 5 m M . La reacc ion fin a liza con la ad ic io n de 1,5 m l de C H C h: C H ³ O H (1: 2), seg u ido de la ad ic io n de agua para lle va r el vo lu m e n acu oso to ta l a 0 ,9 ml. D espues , la fase o rg a n ica se an a liza po r c ro m a to g ra fia de capa fina ju n to con pa tro ne s a d ecu ad os , u tilizan do p lacas p re p a ra d a s con 40 g de ge l de s ilice H su sp e n d id o en 95 m l de te tra b o ra to de sod io 1 m M . El s is tem a d iso lve n te co n s is te en C H C l.³ :C H ³ O H :H ² O (95 :35 :5 ).

If you have a reaction to a reaction that has 10 pl of uncle, it is more than 10 pl of the first letter that is not the minimum in 3 mM, 25 pl of T ris -HC l 100 µM (pH 7.0) and 5 pl of 5 µM EDTA. The end-to-end reaction with the addition of 1.5 ml CHC h: CH ³ OH (1: 2), following the addition of water to bring the water volume bear to ta 0.9 ml. Then, the single phase is analyzed by the thin-layer film with the help of sad parents, using the lacquers for 40 g. of silica gel H its sp end id in 95 ml of 1 mM sodium tea. The s is tem ad iso lve n te co ns is te in CHC l. ³ : CH ³ OH: H ² O (95: 35: 5).

Ensayo de sustrato de proteasa:

Protease Substrate Assay:

S e p re pa ra una m ezc la de reacc ion que con tien e 10 pl de e n z im a m ezc lad a con 800 pl de T ris -H C l 12,5 m M (pH 8 ,0 ) que co n tie n e T rito n X -100 al 0 ,25 % y 10 |jl de C b z -P h e -O N p d isu e lto en D M S O . El p -n itro fen o l libe ra do a tra ve s de la esc is ion del sus tra to se m ide c o n tro la n d o la a b so rb a n c ia a 405 nm.

S ep re for a reaction mixture with 10 pl of enzyme with 800 pl of T ris -HC l 12.5 mM (pH 8, 0) which has T rhythm X -100 at 0.25 % and 10 | jl of C bz -P he -ON pd isu e lto in DMSO. The phenomenon frees up through the analysis of its treatment with the open source at 405 nm.

Ensayo de hidrolisis de acil-PNP graso:

Acyl-PNP fatty hydrolysis assay:

En p r im e r lugar, se p re pa ra una so luc io n de rea c tivo que co n tie n e T riton X -100 al 2 % en fo s fa to de sod io 50 m M , pH 7 ,0 y C i² -p -n itro fe n o l (a c il-P N P ) 10 m M en ace tona . D espues, se p re pa ra una so luc io n de tra b a jo de C ¹² -P N P m e zc la n d o 600 j l de C ¹² -P N P 10 m M en un ta m p o n fo s fa to de 9 ,4 ml.

First, there is a reagent solution that has a 2% X-100 rhythm in 50 mM sodium pH, pH 7.0 and C i ² -p -n itro fe nol (ac il-P NP) 10 μM in acetone. Then, a work solution of C ¹² -PNP was carried out with the 600 jl node of C ¹² -PNP 10 mM in a size of 9, 4 ml.

El en sayo se rea liza anad ien do , a cada poc illo de una p laca de 96 poc illos , 40 j l de la so luc io n de tra b a jo de ac il-PN P , seg u ido de la rap ida ad ic ion de 40 j l de enz im a . La so luc ion se m ezc la du ran te 15 se g un do s y el cam b io de a b so rb a n c ia se lee a 405 nm en un le c to r de p lacas de m ic ro titu lac ion a una te m p e ra tu ra de 25 °C.

The essay is carried out by adding, to each well of a 96-pound plate, 40 jl of the work solution of the National Park, according to the rapid transit ad ic ion of 40 jl of enzyme im. The solution shows the 15th week and the exchange rate is read at 405 nm in a reading of the lacquer plates at one time. it measures your ra at 25 ° C.

Formacion de ester a partir de tioester:

Formation of ester from thioester:

S e p re p a ra una m e zc la de rea cc io n que co n tie n e e n z im a tio e s te ra s a 1,5 jM , m ir is to il-C o A 100 jM , m e tan o l a l 10 % (v /v ) y fo s fa to de sod io 50 m M , a un pH de 7,0. La m ezc la de reacc ion se in cub a du ran te 1 hora a 20 °C y fina liza con la a d ic io n de H C l 1 N pa ra d is m in u ir el pH a a p ro x im a d a m e n te 2 ,5. La m ezc la se e x tra e con un m ism o vo lu m e n de a ce ta to de e tilo y la can tidad de e s te r g ra so p ro d u c id a se d e te rm in a m e d ia n te G C -M S u o tros m e to d o s e s ta n d a r ta le s com o G C -F ID , LC -M S , o c ro m a to g ra fia en cap a fina.

S ep re para ra me a reaction area that has a 1.5%, it is 10% (100%). / v) and fo s fa to sodium 50 mM, at a pH of 7.0. The reaction mixture is covered in 1 hour at 20 ° C and finely with the addition of HC l 1 N pa ra d is m in u go to pH aap ro x im adamen te 2, 5. The m ezc is extracted with a very low volume and the amount of money will be given only by the GC in the middle of GC -Mo tros m e to dos ta ndar ta le s com o GC -F ID, LC -MS, oc o mary in fine cap.

Formacion de ester a partir de ester:

Formation of ester from ester:

S e p re pa ra una m ezc la de reacc ion que con tiene e n z im a tio e s te ra sa 1,5 jM , la u ro il-C o A 300 jM , m e tan o l al 10 % (v /v ) y fo s fa to de sod io 50 m M , a un pH de 7,0. La m ezc la de reacc ion se in cub a du ran te 1 hora a 20 °C y fina liza con la a d ic io n de H C l 1 N pa ra d is m in u ir e l pH a a p ro x im a d a m e n te 2 ,5. La m e zc la se e x tra e con un m ism o vo lu m e n de a ce ta to de e tilo y la can tida d de lauril e s te r p ro du c ida se de te rm in a m ed ia n te G C -M S u o tros m e tod os e s ta n d a r ta le s com o G C -F ID , LC -M S , o c ro m a to g ra fia en cap a fina.

S ep re for a reaction period that has an enzyme of 1.5 jM, the u-il C or A 300 jM, so ol 10% (v / v) and fo s fa to sodium 50 mM, at a pH of 7.0. The reaction mixture is covered in 1 hour at 20 ° C and finely with the addition of HC l 1 N pa ra d is m in u go to pH aap ro x im adamen te 2, 5. The media is extracted with a minimum volume of money, and the amount of money is given to you in the year in GC. -My other methods is common as GC-F ID, LC-MS, with a high level of trust.

Las e xp res ione s ce lu la " tra n s fo rm a d a " o "tra n s fo rm a d a de m an e ra estab le ", se re fie ren a una ce lu la que tien e una se cu e n c ia de p o lin u c le o tid o s no na tiva (h e te ro lo g a ) in te g ra da en su g e no m a o com o un p lasm ido ep iso m a l que se co n se rva d u ran te al m en os dos ge ne rac io ne s .

The e xp res ione s ce lu the "tra ns fo rm ada" or "tra ns fo rm ada de m an e ra esta le", are referred to a ce that has a se c Po lin uc le o tid os not na tiva (he ro lo lo ga) in te g ra da in its ge no mao com o a p plasmid gone ep iso bad that co n rva du ran te to me in two o'clock ge ne rac io ne s.

La e xp res ion "p ro te in a tra n s p o rta d o ra " se re fie re a una p ro te ina que fac ilita el m o v im ie n to de uno o m as co m p u e s to s de n tro y /o fue ra de un o rg an ism o u o rganu lo . En a lgu nos aspec tos , el h o sp e d a d o r de p roducc ion e xp resa ra fu n c io n a lm e n te una se cu e n c ia de A D N exo g e n a que cod ifica una p ro te ina tra n s p o rta d o ra de ca se te s de un ion a A T P (A B C , ATP-Binding Cassette) de m odo que d icho h o s p e d a d o r e xp o rte el d e riva d o de ac ido g ra so al m ed io de cu ltivo . Las p ro te inas tra n sp o rta d o ra s de A B C se en cue n tran en m uchos o rg an ism os, ta le s com o Caenorhabditis elegans, Arabidopsis thaliana, Alcaligenes eutrophus (p o s te rio rm e n te ren om b rado com o Ralstonia eutropha) o R h od ococcu s e ry th ro p o lis . C o m o e je m p lo s no lim ita tivo s de p ro te in a s tra n s p o rta d o ra s de A B C se in c luye n C E R 5, A tM R P 5 , A m iS 2 y A tP G P 1. En un a sp e c to p re fe rid o , la p ro te in a tra n s p o rta d o ra de A B C es C E R 5 (p o r e jem p lo , A Y 734542 ). En o tros asp ec tos , la p ro te ina tra n sp o rta d o ra es una p ro te ina de sa lida s e le cc io n a d a de: A crA B , T o lC o A c rE F de E. coli o tll1618 , t ll1619 y t ll0139 de Thermosynechococcus elongatus BP-1. En asp e c to s ad ic io na le s , la p ro te ina tra n s p o rta d o ra es una p ro te in a tra n s p o rta d o ra de ac ido s g ra so s (F A T P , fatty acid transport protein) s e le c c io n a d a de Drosophila melanogaster, Caenorhabditis elegans, Mycobacterium tuberculosis o Saccharomyces cerevisiae o de cu a lq u ie ra de las F A TP de m am ife ros con oc idas en la m a te ria . Las p ro te inas tra n sp o rta d o ra s son utiles, por e jem p lo , para m e jo ra r la sec re c io n o libe ra c io n de p ro du c to s que de otro m od o no son cap ace s de se c re ta r e sp o n ta n e a m e n te el p roducto . T am b ien son u tiles cu a n d o las ce lu las ho sp e d a d o ra s g e n o m a n ip u la d a s son capaces de s e c re ta r o lib e ra r e s p o n ta n e a m e n te el p roducto , a u nq ue lo libe ren le n tam e n te o de m an e ra in com p le ta . En esas c ircu n s ta n c ia s , las p ro te inas tra n s p o rta d o ra s pu ed en m e jo ra r la sec re c io n al a c e le ra r la e ta pa de sec re c io n o lle va r a cab o la sec re c io n hasta su fina liza c io n .

The "p ro te in a tra nspo rta do ra" expression refers to a project that facilitates the movement of one or more other persons and / or it was ra of an o rg an ism ouo rganu lo. In some respects, the ho age of production and xp resa ra fu nc io na lm in which a sign of exogenous DNA coding for a road through the capital You have an ATP (ABC, ATP-Binding Cassette) ion of the method that said host xp or rt of the riva do de r ado gone ra to the quantitative environment. The first steps of ABC are found in many countries, such as Caenorhabditis elegans, Arabidopsis thaliana, Alcaligenes eutrophus (post r r r in te ren om b rado com or Ralstonia eutropha) or R h od ococcu se ry th ro po lis. How do you think that is not limited to ABC's activities? It is included in CER 5, A RP RP 5, A 2, and GP GP 1. In a A sp ec to p re fe rid o, the p r te in through nspo rta do ra of ABC is CER 5 (pore jem p lo, AY 734542). In other cases, the plan for the transfer is an exit from which: A crA B, T o lC or A c rE F de E. coli or tll1618, t ll1619 and t ll0139 of Thermosynechococcus elongatus BP-1. In asp ec to s ad ic io na le s, the p ro te na tra nspo rta do ra is a p ro te in th e trans rs do s ra ra so s (FATP, fatty acid transport protein) is He has nothing to do with Drosophila melanogaster, Caenorhabditis elegans, Mycobacterium tuberculosis or Saccharomyces cerevisiae or with one of the most common AF AFs that occur in the world. P ro te ines tra n sp o rta do ra s are useful, for example, for the sake of the sector, they do not release them from other countries or they are not captains of se c re ta re sp on ta neamen te the product. They are also useful when they have been released by the sgenoman ip u the days they are able to respond directly to the product, even if they release it at the same time. I o m in e p ra in com p le ta. In these cases, the institutions that will be able to improve the sec- ond section of the sector will not be able to do so. It will be up to the end until it is fine.

En el p re sen te d o cum e n to , los te rm in o s "va ria n te " y "m u ta n te " se u tilizan in d is tin tam en te .

In the past, the rm in o s "va ria n te" and "mu ta n te" are used in d is tin tam in te.

Un "ve c to r" se re fie re a una co n s tru cc io n p o lin u c le o tid ica d ise n a d a pa ra in tro d u c ir p o lin u c le o tid o s en uno o m as tipo s de ce lu las . Los ve c to re s inc luyen ve c to re s de c lonac ion , ve c to re s de exp res ion , ve c to re s lanzadera , p lasm idos, ca se te s y s im ila res . En a lg u n o s aspec tos , la co n s tru cc io n p o lin u c le o tid ica co m p re n d e una s e cu e n c ia p o lin u c le o tid ica que co d ifica una tio e s te ra sa (p o r e jem p lo , una t io e s te ra s a p re cu rso ra o m ad u ra ) que esta un ida o p e ra tiva m e n te a una p ro secu enc ia a d ecu ad a (p o r e jem p lo , una p ro se cu e n c ia se c re to ra ) cap az de e fe c tu a r la exp res ion del po lin u c le o tid o o gen en un h o sp e d a d o r adecuado .

A "ve c to r" refers to a co ls tru cc io npo lin uc le or tidica d ise nad para ra in tro duc go po lin uc le tid os in one or more other types of ce lu las. The ve c to re s inc see ve c to re s of the canvas, see the expiration, see the shuttle, plasmids, ca se s s and im ila res. In some respects, the co-op ct npo lin uc le or tidica co mp re nde a co n tio c a l p l e ct o c u l le or tidica that an uncle co ifies is te ra sa (pore jem p What, one guy is te sap re qu rso ra om ad u ra) which is an operative way only a se se se c on ad equ ad a (pore jem p lo, a p ro se cu en ia se c re to ra) chapter of the faith with the expiration of the po u u le le tid oo gene in a suitable age.

U na "ce ra " es una su s ta n c ia que com p ren de , al m en os en parte, es te res g rasos. En d e te rm in a d o s aspec tos , un e s te r g raso tien e un lado A y un lado B, c o m p re n d ie n d o cada uno de e llos ca d e n a s de c a rb o n o m e d ia n a s y la rgas. A d e m a s de es te res grasos, una cera puede co m p re n d e r o tros co m p on en te s . P o r e jem p lo , una ce ra puede co m p re n d e r h id ro ca rbu ros , es te res de este ro l, a lde h ido s a lifa ticos, a lcoh o le s , ce tonas , be ta -d ice ton as , tr ia c ilg lic e ro le s y s im ila res . N o rm a lm e n te , una ce ra es un so lido a te m p e ra tu ra am b ie n te , po r e jem p lo , a 20 °C.

A "ce ra" is one of the only things that you can buy from, at least in part, this is the reason. In terms of specific aspects, one of them has a side A and a side B, each one of which is considered to be nasty and nasty. In addition to these fatty substances, a wax may also contain other components. P o jem p lo, a ce can be co r rd nderh id ro ca rbu ros, this is about this r l, to the sa l r a ry, to lcoh or le s, ce tonas, be ta-d ice ton as, tr ia c ilg lic e ro le sys im ila res. No rm a lm in te, a wax is a solid to you, too, at least 20 ° C.

"T ip o s ilves tre " s ign ifica , en el con tex to de un gen o una p ro te ina , una se cu e n c ia de p o lin u c le o tid o s o de p ro te inas que se p ro du ce en la na tu ra leza . En a lgu nos aspec tos , la secu en c ia de t ip o s ilves tre se re fie re a una secu en c ia de in te res que es un pu n to de pa rtida para el d iseno de p ro te inas m ed ia n te in ge n ie ria gene tica .

"T ip os ilves tre" s means, in the context of a gene or a group, a group of people who has been able to protect them. In nature . In this regard, the sectoral section of the IPs refers to a section of interest that is a starting point for the design of the project. I am an engineer in genetics.

Produccion de derivados de acidos grasosProduction of fatty acid derivatives

De acu e rd o con un asp ec to de la p resen te d ivu lg ac ion , las nuevas tio e s te ra s a s se exp resa n en una ce lu la h o sp e d a d o ra que es cap az de tra n s fo rm a r una fue n te de ca rb on o en un de riva do de ac ido graso . La d ivu lg ac ion se re fie re a dos a sp e c to s d is tin tos : (1) el d e scu b rim ie n to de que pu ede u tiliza rse una t io e s te ra s a m u tan te para o p tim iza r y /o "d ise n a r" una com p o s ic io n de de riva do de ac ido g ra so para h a ce r que d ichas co m p o s ic io n e s sean m as u tiles y que d ife re n te s m u ta c io n e s p ro po rc ion en d ife ren te s p ro p ieda de s d iana ; y (2) el d e scu b rim ie n to de que la t io e s te ra sa ac tu a ra en una ce lu la h o sp e d a d o ra re co m b in a n te para p ro d u c ir d ire c ta m e n te p ro du c to s de e s te r graso, sin la p re se n c ia de una e n z im a ce ra s in ta sa o e s te r s in tasa .

Based on an aspiring idea, the new uncle is only in a new house that is now a captain of his work. A source was a source of a fatty acid source. The decision refers to two questions: (1) that of the fact that an uncle can be used is the sam so much for you. It is a "d ise n ar" a com ivation of a riva do de r a c a g ra so so that these com- positions are more useful and that they should be i smu ta c io nesp ro po rc ion in d ife ren te sp ro p ieda de sd iana; and (2) that of scu b rim ie n to the fact that the uncle is in one place at a time of the year in order to go to the direction of the men It is fatty, without the presence of an enzyme that is in place or at the rate.

De a cu e rd o con un aspecto , las com p o s ic io n e s de de riva dos de ac ido s g rasos p ro du c ida s segun los m e todos, los ve c to re s y las ce lu la s del p re sen te do cum e n to , tie n e n p ro p ie d a d e s m o d ifica d a s o a lte rad as en com p a ra c io n con los d e riva d o s de ac ido s g rasos p ro du c ido s u tilizan do ce lu las ho sp e d a d o ra s que no com p ren den las va r ia n te s de t io e s te ra sa de la d ivu lg ac ion . P o r e jem p lo , com o ta m b ie n se d e sc rib e en el p re sen te d o cum e n to , u tilizan do las tio e s te ra s a s de la p re sen te d ivu lg ac ion , es pos ib le d e s a rro lla r p ro ceso s de fa b rica c io n que p ro du zcan de riva dos de ac ido s grasos, que, en com p a ra c io n con un p ro ceso s im ila r que im p liq ue una t io e s te ra sa de t ip o s ilves tre , tien e pe rfile s de com p o s ic io n a lte rados , po r e jem p lo , po rcen ta jes a lte rad os de una serie de, o de un g ru po ac ilo de long itud de cadena de ca rb on o espec ifica , g ru po s ac ilo sa tu ra do s o in sa tu ra dos , pos ic ion de in sa tu ra c ion es , g rupos ac ilo ram ificados , pos ic ion de ram ificac io n , g ru po s h id ro x ilo -ac ilo , g ru po s ce to -ac ilo , p ropo rc ion de es te res o ac idos g ra sos lib res en el p roducto , p ropo rc ion de de riva dos de ac ido s g rasos de cadena corta (p o r e jem p lo , C8, Cg, C ¹⁰ , C ¹¹ , C ¹² , C ¹³ y /o C ¹⁴ ) fren te a cadena la rga (p o r e jem p lo , C ¹⁵ , Cia, C ¹⁷ , C ¹⁸ , C igy /o C ²⁰ ), o re n d im ie n to de de riva dos de ac ido s g rasos. P o r co n s ig u ie n te , pu eden g e n o m a n ip u la rs e p ro d u c to s con d ive rsa s p ro p ie d a d e s de sea b le s , de ta l m an e ra que te n g a n nu m ero s de cetano , ind ices de oc tano , e s ta b ilid a d ox ida tiva , lub ric idad , pun tos de in flam ac ion , v isco s id a d , p u n tos de e b u llic ion , pu n tos de fus io n , p u n tos de flu idez , p u n tos de e n tu rb a m ie n to , pu n to de o b s tru cc io n de l filtro en frio , ca ra c te ris tica s de flu jo en frio , a ro m a tic id a d y /o nu m ero s de yo d o op tim iza dos .

With respect to one aspect, the riva two-year-old composites according to the methods, see them and see them in the p I feel that I have a lot of time, I have no idea that I want to see the radius in comparison with those of the two in the second century, but they have branded it as the last hours. Those who do not understand the values of uncle are the members of the government. P o jem p lo, as well as sc rib e in the p re sen te do cum in to, u use the uncle s teas sas de la p re sen te d ivu lg ac ion, is After that, the most important factor in the process of making two fatty acids, which, in comparison with a small number of people, is that im p liq ue a un io es te ra sa de t ip os ilves tre, te e sa rfile s de com pos ic io na lte rados, po re jem p lo, po r ta ta jes a lte rad os of a series of , or of a specific group of long-chain of specific chain, it is also available for you, for your position, it is your position. g branched acyls, branching position, g po po sh id ro x ilo -ac ilo, g ru po s ce to -ac ilo, p ropo rtion of this acid or acids g ra es lib lib in the product, p ropo rc ion de de riva dos de ac a sg short chain (for example, C8, Cg, C ¹⁰ , C ¹¹ , C ¹² , C ¹³ and / or C ¹⁴ ) stop the chain rga (pore jem p lo, C ¹⁵ , Cia, C ¹⁷ , C ¹⁸ , C igy / o C ²⁰ ), or re r nd im e n of de riva dos de acido sg rasos. For example, they can genoman ip u la rs ep ro duc to s with d ive rsa sp ro p ie dades de sea b le s, so l n an e ra that n den u number s Of cetane, indexes of the ocean, it is also important, lubicity, flash points, video ID, boiling points, melting points Fluid points, points in your life, cold filter blockage, cold flow, high temperature id ady / o num m ero s de yo do op tim iza dos.

Los d e riva d o s de ac ido s g ra sos son u tiles com o, o com o c o m p o n e n te s de, b io co m b u s tib le s y p ro du c to s qu im icos e sp ec ia lizad os . Los de riva d o s de ac ido s g rasos y los p ro du c to s fa b rica d o s a pa rtir de e llos inc luyen com b ustib les , ad itivo s de com b ustib le , a le a c io n e s de com b us tib le , d e te rg e n te s y te n s io a c tivo s , co m p le m e n to s nu tric iona les , po lim eros , ree m p la zos de pa ra fina , lu b rica n tes , d iso lve n tes , p ro du c to s para el cu ida do pe rsona l, ad itivo s de p ro ce sa m ie n to de caucho , in h ib id o res de co rros ion , e m u ls io na n tes , p las ticos, te jido s , cosm e ticos , p ro du c to s de pape l, recu b rim ien to s , liqu id os para la m e ta lu rg ia , d ie le c tricos , en g ra sa n te s y e m o lie n tes . Los m e to d o s y co m p o s ic io n e s d e s v e la d o s en es te d o cu m e n to p e rm iten la p ro d u cc io n de d e riva d o s de a c id o s g ra so s con p u n tos de ram ificac io n , n ive les de sa tu ra c io n y long itud de la cad en a de ca rbono , pa rticu la res . Los m e tod os y co m p o s ic io n e s de este d o cu m e n to ta m b ie n pe rm iten la p ro du cc ion de una m a yo r p ro po rc ion de es te res g ra sos fren te a o tros p roduc tos , o com o a lte rna tiva , una m e n o r p ro po rc ion de este res g ra so s fre n te a o tros p roductos, de p e n d ie n d o de si es d e se a b le un m a yo r re n d im ie n to p ro p o rc io n a l o po rcen tu a l de es te re s g ra sos o un m e n o r re n d im ie n to p ro p o rc io n a l o po rcen tu a l de es te res grasos. E spe c ificam en te , po r e jem p lo , los m e tod os y c o m p o s ic io n e s de este d o cum e n to pe rm iten la p ro du cc ion de una m a yo r p ro po rc ion de es te res de ac ido s g ra sos fre n te a ac ido s g ra sos lib res, o en o tras pa lab ras, pe rm iten un m a yo r ren d im ie n to p ro p o rc io n a l o po rcen tu a l de e s te re s de ac ido s g ra sos fre n te a ac idos g ra sos lib res. C o m o a lte rna tiva , po r e jem p lo , los m e tod os y com p o s ic io n e s de este d o cu m e n to pe rm iten la p ro d u cc io n de una p ro po rc ion m as pe q u e n a de es te re s de ac ido s g ra sos fre n te a ac ido s g ra so s lib res, cu a n d o no se desean g ra n d e s ca n tid a d e s de es te re s de ac ido s g rasos. A s im ism o , los m e tod os y c o m p o s ic io n e s de este d o cu m e n to pe rm iten la p ro du cc ion de un re n d im ie n to m e jo rad o de de riva d o s de ac ido s grasos.

The diva rives of these actions are useful as, or comm c o m p o n e n s of, b io co m b u s tib le s and p ro du c to s chemicos e sp ec ia lizad os. Those from Riva Dos de Acido Sg Rasos and Los Pro Du C to S FaB Rica dosa rtir de l l Inc include com b ustib les, com adivesivo de com b ustib le, a com ions of com b Us tib le, from te rg in te s and te ns io ac tivo s, co mp le ment to s nu tric iona les, po e eros, ree mp zos de pa ra ra fine, lu b rica n tes, d iso lve In this case, for the first time of the rubber, rubber adhesion, rubber inlets, emu ls io na n tes , p las ticos, i jido s, cosmetics, p ro du c to s de pape l, rec b rim ien to s, liquids for the meta l rg ia, d ie le c tricos, in g ra sa n te syemo lie n tes. The two methods and comm ies nesveve the two in which you are interested in making the p r du du tion of de riva dos de a c id osg ra so s with branching points, Levels of health and long-term care of the cad in a ca rbono, pa rticu la res. The methods and co-ops of this do c men t ta mb ie n pe rm iten in the p ro du tion of a ma me rp ro po rc ion of this res g ra sos fren te ao tros products, or as a matter of course, a minor share of the latter in terms of products, depending on whether it is of a major nature. ie n to p ro po rc io nalo po rc io n this re sg ra son or a lesser re nd im ie n to p ro po rc io n r p io n r p io n r r io n o r r io n po n po r n im n alo n p n p n p n p n p p p n po n po n p n p p po n po n p n I spe c ticam in te, po re jem p lo, the methods and components of this do cum in to pe rm it in the p ro du ction of a mother and rp ro po rion of this te In this way, you will be freed in this way, or in or after words, but in a better way you will not be able to do it. This is what you say in the future. How do I know how to do it, the methods and com positions of this do in order to do a better job than This is the reason why people are still free, when they don't want to know what they are going to do next. In addition, the methods and components of this document have the effect of making a reimbursement of a riva two of ac fatty

C o m o e je m p lo s no lim ita tivo s de m ic ro o rg a n ism o s que pueden u tiliza rse com o ho sp e d a d o re s de p ro du cc ion para p ro d u c ir de riva dos de ac idos g ra sos se inc luyen c ia no bac te rias , a lgas, bacte rias , le vad u ras u hongos fila m en to sos . O tro s e jem p los no lim itan te s de ho sp e d a d o re s de p roducc ion ad e cu a d o s in c luyen ce lu las ve g e ta le s , an im a le s o hum anas.

How do I know what is not limited to me or what can be used as a source of protection for two ducts of two acids? If they are included, they will not bac rias, then, bacte rias, vad u ras u fungi row m in all. O s t he jem p not limited to ho sp e d a d o re s de p roduction io ad e cu a d o s in c lu lu c ve lu e ve le g e ta le s, an im a le or o hu anas.

Los h o sp e d a d o re s de p ro du cc ion d e sc rito s en es te do cu m e n to , pu ed en p ro d u c ir a lco h o le s (de cad en a corta , cad en a la rga , ram ificada o insa tu ra da ). D ichos a lcoh o le s pueden u tiliza rse d ire c ta m e n te com o co m b u s tib le s o pueden u tiliza rse para c re a r un e s te r graso. Los es te res g rasos, so los o en com b in ac io n con o tros de riva dos de ac idos g ra sos de sc rito s en el p resen te do cu m e n to , ta m b ie n son u tiles com o co m b u s tib le s o com o c o m p o n e n te s de com b us tib les .

Ho sp edado re s de p ro du cc ion de sc rito s en es te do cu men to, pu ed en p ro duc go to lco ho le s (from cad to short, cad to to rga, ram ified or insa your ra). Sayings to lcoh or s can be used as a common source or can be used to create a fatty substance. These are the only ones that are shared with others of two major acidic acids in the country where they are also useful. How do you know how to make them?

De m an era s im ila r, los h id ro ca rb u ro s p ro d u c id o s a p a rtir de los h o sp e d a d o re s de p ro du cc ion d e sc rito s en el p re sen te do cu m e n to , pu ed en u tiliza rse co m o b io co m b u s tib le s o com o c o m p o n e n te s de b io co m b u s tib le s . D ichos co m b u s tib le s ba sad os en h id ro ca rb u ro s pueden d ise n a rse pa ra que con teng an co n te n e r pu n tos de ram ificac io n , g ra do s de sa tu ra c io n d e fin id o s y lo ng itude s de ca rb o n o esp ec ificas , u tilizan do las en se n a n za s p ro p o rc io n a d a s en este do cu m e n to . C u a n d o se u tilizan com o b io co m b u s tib le s so los o en co m b in a c io n con o tros d e riva d o s de ac ido s grasos, los h id ro ca rbu ros pueden co m b in a rse con ad itivo s a d ecu ad os u o tros co m b u s tib le s tra d ic io n a le s (p o r e jem p lo , a lcoh o le s , d iese l p ro ce d e n te de tr ig lic e r id o s y co m b u s tib le s ba sa d o s en pe tro leo).

In the same way, they are the same as the duo id osapa rtir de los ho sp edado re s de p ro du cc ion de sc rito s in the p re sen te do cu men to It is possible to use co mob io co mbus tib le so as to compose te b io co mbus tib le s. These co-busses can be found in other countries so that they may have ramifications, branches of interest Your aim is for the purposes of this type of use, using the signs indicated by this group. When they are used as common use only with others or with other types of fatty acids, those who are able to take care of them itivo sad ecu ad os uo tros co mbus tib le s tra d ic io na le s (pore jem p lo, lcoh o le s, say lp ro ce den te de tr ig lic e r id osy co mbus tib le s ba sa two in pe tro tro).

El nu m ero de ce ta n o (N C ), la v isco s id a d , el pun to de fu s io n y el c a lo r de com b us tio n de d ive rso s es te res g ra sos se han ca ra c te riza d o en K no the , Fuel P roce ss in g T e ch n o lo g y 86 :1059 -1070 , 2005. Un h o sp e d a d o r de p roducc ion puede g e n o m a n ip u la rs e para p ro d u c ir cu a lq u ie ra de los e s te re s g ra sos d e sc rito s en Knothe, u tilizan do las e n se n a n za s p ro p o rc io n a d a s en este docum en to .

The number of ceta no (NC), the v isco s id ad, the point of fu s io n and the com ca r r of the com ive of the ive rso s these are the rest It is in K no the, Fuel P roce ss in g T e ch no lo gy 86: 1059 -1070, 2005. A ho p age of p roduction can genoman ip u the rs e for p ro duc Go to one of the re sg ra es of sc rites in Knothe, use the signs indicated in this document in this document.

I. Produccion de derivados de acidos grasos y modificaciones para mejorar la produccion/rendimiento El h o s p e d a d o r de p ro du cc ion u tilizado para p ro d u c ir a c il-C o A y /o d e riva d o s de ac ido s g ra sos pued e m o d ifica rse de fo rm a re co m b in a n te para que in c luya se cu e n c ia s de p o lin u c le o tid o s que sob re e xp re se n pep tidos . P o r e jem p lo , el h o s p e d a d o r de p roducc ion puede m o d ifica rse para a u m e n ta r la p roducc ion de a c il-C o A y re d u c ir el ca ta b o lism o de d e riva d o s de ac ido s g ra sos y de p ro du c to s in te rm e d io s en la ruta b ios in te tica de ac idos grasos, o para re d u c ir la in h ib ic ion po r re tro a lim e n ta c io n en pun tos e sp e c ifico s de la ru ta b ios in te tica de ac idos grasos. A d e m a s de m o d ifica r los ge ne s d e sc rito s en el p re se n te do cu m e n to , se pu ed en d e d ic a r re cu rso s c e lu la re s a d ic io n a le s pa ra s o b re p ro d u c ir ac ido s g rasos. P o r e jem p lo , las ru tas de lac ta to , su cc in a to y /o ace ta to p u ed en a te nua rse , y la a ce til-C o A ca rb ox ilasa (acc) puede sob re e xp re sa rse . Las m od ifica c io n e s en el h o sp e d a d o r de p ro du cc ion de sc ritas en el p resen te d o cu m e n to pu ed en re a liza rse a tra v e s de a lte ra c io n e s g e no m icas , ad ic ion de s is te m a s de e xp res ion reco m b in an te , o co m b in a c io n e s de las m ism as. P o r e jem p lo , una o m as t io e s te ra sa s en d o g e n a s de un h o s p e d a d o r de p roducc ion p a rticu la r pueden m o d ifica rse u tilizan do te c n ic a s ad e cu a d a s de m od o que el t io e s te r m u ta n te te n g a al m en os una p ro p ieda d a lte ra d a en c o m p a ra c io n con el p re cu rso r de t io e s te ra s a e n do ge no , o de m o d o que la ce lu la h o sp e d a d o ra m u e s tre al m en os una p rop iedad a lte rada , en com p a ra c io n con la m ism a ce lu la ho spe d a d o ra an tes de que este su je ta a las e ta pas de a lte rac ion genom ica .

I. Production of fatty acid derivatives and modifications to improve production / yield The host of the use or use for the benefit of the A-C or A and / or riva two of the acid can be followed. Sign up for help in order to see how much you need to know. P o jem p lo, the host of modification can be modified to increase the production of ac-C or A and reduce the cost of the two-year accrual. In this way, in the path of fatty acids, or in order to reduce the risk to reduce the limitation in this area. In points of the ecological route of fatty acids. In addition to modifying the sc rites in the president, it is possible to recount what happened to the sad sadness of the country. re p ro duc go ac se sg rasos. For example, lacta to routes, their cc in a to and / or aceta to pu ed en a nua rse, and the ail til-C or A ca rb ox ilasa (acc) can sob re e xp re sa rse. The metrics in the ho age of the sc ritas of the r p in the past are made in the process of being carried out through the past In addition, there is more than one of the most important reactions, or commbitions of the same. P o jem p lo, one or more of them is in the days of a host of patriotic production that can be modified or used as part of the process that the uncle will ruminate you in m a p p p p eda da lte ra da in comparison with the p r r o of un tio es te ra sa sa do do ge no, or so that the card has been shown at a time to one year, in comparison with the same time as the letter given before it is your boss at the age of the genomic ration.

Las ru tas b ios in te ticas de ac idos g ra sos im p lica da s se ilus tran en las FIGS. 2-5. Los s ig u ie n tes su b a p a rta d o s A -G de sc rib en las e ta pas en estas rutas. D ive rsas e n z im a s ca ta lizan d ive rsas e ta pas en la ruta. P o r con s ig u ie n te , cada e ta pa es un po s ib le lu g a r para la so b re e xp re s io n de l gen pa ra p ro d u c ir m as una o m as e n z im a s y, po r lo tan to , im p u ls a r la p ro du cc ion de m as ac idos g ra sos y de riva d o s de ac ido s grasos. T am b ien pueden an a d irse genes que cod ifiq uen las en z im a s ne cesa rias para la ru ta de m an e ra re co m b in a n te a un h o s p e d a d o r de p ro du cc ion que ca re zca de d ich a s enz im as. P o r u ltim o, para a u m e n ta r la p ro du cc ion de los p ro du c to s de sea do s , las e ta pas que co m p e tiria n con la ru ta que co n d u ce a la p ro du cc ion de ac ido s g ra sos y de riva dos de ac ido s g ra sos pueden a te n u a rse o b loquea rse .

The theoretical routes of acidic acids are implied in FIGS. 2-5. The experts in their b a p a rta d o s A -G of sc rib in the states on these routes. D ive rsas e n z im a s ca ta lizan d ive rsas e ta pas on the route. For example, each country is a place to go for the purpose of the gene to go beyond one or more other enzymes. Therefore, it is possible to increase the fatty acids and the two fatty acids. You can also add genes that code if you need them for the route of the journey to a host that falls from d ich as enzy im as. For the purpose of increasing the p r du du tion of the p ro du c to s of sea do s, the countries that would cope with the route that leads to the p ro du c ion of ac s s sos dos dos y y y y dos dos pueden pueden pueden pueden pueden pueden pueden pueden pueden pueden pueden pueden pueden pueden pueden pueden pueden pueden pueden pueden pueden

De acu e rd o con las d ivu lg a c io n e s de este do cu m e n to , un exp erto en la m a te ria puede u tiliz a r las tio e s te ra s a s de la d ivu lg a c io n para p re p a ra r m ic ro o rg a n ism o s que p roduzcan d e riva d o s de ac ido s g ra sos y para fa b ric a r d ive rsos d e riva d o s de ac ido s g ra sos u tiliza n d o d icho s m ic ro o rg a n ism o s , en dond e d icho s de riva dos de ac ido s g ra sos tienen p ro p ieda de s a lte rad as . T a m b ie n es pos ib le p re p a ra r m ic ro o rg a n ism o s que p ro du zcan d icho s d e riva d o s de ac idos g ra sos de m a n e ra m as e ficaz al te n e r los n ive les de ren d im ien to , p ro du c tiv ida d o titu lo desea do s , du ran te las fe rm e n ta c io n e s .

In accordance with the provisions of this document, an expert in the ma- rior can use the uncles as part of the decisions to pay for him. ra rm ic ro or rg an ism os p rdudu de riva dos de acido sg ra sos and for fa b ric ard ive rsos de riva dos de acido sg ra sos u use ndod icho sm ic ro o rg an ism os , in which riva dos de riva dos de ac ido sg ra sos have p ro p ieda de sa lte rad as. It is also possible that there are more important reasons that two of the most acidic acids are more efficient in terms of levels. If you wish to do so, you may wish to do so, if you have faith in them.

A. Acetil-CoA - Malonil-CoA a Acil-ACP

A. Acetyl-CoA - Malonyl-CoA to Acil-ACP

La ac ido g ra so s in ta sa (F A S ) es un g ru po de pe p tid o s que c a ta liza el in ic io y el a la rg a m ie n to de las ca d e n a s de ac ilo (M a rra kch i e t al., B io che m ica l S oc ie ty , 30 :1050 -1055 , 2002 ). La p ro te ina tra n s p o rta d o ra de ac ilo (A C P ), ju n to con las en z im a s de la ru ta FAS, con tro la n la long itud , el g ra do de sa tu ra c io n y la ram ificac io n de los ac ido s grasos p ro du c ido s . Las e ta pas de es ta ru ta estan c a ta liza d a s po r e n z im a s de las fa m ilia s de genes de la b ios in tes is de ac ido s g ra sos (fab) y a c e til-C o A ca rb o x ila sa (acc). D e pe nd ien do de l p ro du c to deseado , uno o m as de estos genes pueden e s ta r a te n u a d o s o sob ree xp resa do s .

The ac i g ra so s in ta sa (FAS) is a group of credits that can be used for the beginning and the most important of the acyl chains (M a rra kch iet al., B io che m ica l S oc ie ty, 30: 1050-1055, 2002). The plan is for you through the Acroport (ACP), along with the areas of the FAS route, including the long route, the group of employees. n and the ramification of fatty acids for the past. The stages of this route are categorized by the factions of the genes of the genesis of the sg ra sos ( fab) lies til-C or A ca rb ox ila sa ( acc). Due to the desired desire, one or more of these genes may be nuanced as well as possible.

I. R u ta b ios in te tica de ac ido s g rasos: a ce til-C o A o m a lo n il-C o A a ac il-A C P

I. R u ta b ios in the art of ac ect cs in general: a ce til-C or A or m a n il-C or A a ac il-A C P

La ru ta b io s in te tica de ac ido s g ra sos en el h o s p e d a d o r de p ro du cc ion u tiliza la a ce til-C o A y m a lo n il-C o A precu rso ras . Las e ta pas de esta ru ta estan ca ta liza d a s po r en z im a s de las fa m ilia s de genes de la b ios in tes is de ac ido s g rasos (fab) y a c e til-C o A c a rb o x ila sa (acc). E sta ru ta se de sc rib e en H eath e t al., P rog. L ip id Res., 40 (6 ):467 -97 , 2001. La a ce til-C o A ca rb o x ila sa (Acc, una e n z im a con m u ltip le s su b u n id a d e s co d ifica d a po r cua tro genes d is tin tos , accABCD) ca rb o x ila la a ce til-C o A para fo rm a r m a lon il-C o A . La m a lon il-C o A : A C P tra n s a c ila s a (F a b D ) tra n s fie re el g ru po m a lo n a to a A C P para fo rm a r m a lon il-A C P . D espues, se p roduce una reacc ion de con d e n sa c io n , donde la m a lo n il-A C P se fu s io n a con la ace til-C o A , dando com o resu ltad o la p -ce to ac il-A C P . La p -ce to a c il-A C P s in ta sa III (F a b H ) in ic ia el c ic lo FAS, m ie n tras que la p -ce to a c il-A C P s in ta sa I (F a b B ) y la p -ce to a c il-A C P s in ta sa II (F abF ) es tan im p lica d a s en c ic los pos te rio res .

The pathway in the art of safekeeping is in the host of the service or uses the C-A or C y A il C-C or A pre c rs o ras. The stages of this route are categorized by the faculties of genes of the genesis of national relations ( fab) lying til-C or A ca rb ox ila sa ( acc). This route is discussed in H eath et al., P rog. L ip id Res., 40 (6): 467-97, 2001. The company is C-ox or A ca rb ox ila sa (Acc, an enzyme with more than one bun id ades code For four different genes, accABCD) ca rb ox ila la a ce til-C or A for fo rm weapon lon il-C or A. The longest il-C or A: ACP tra nsac ila sa (F ab D) tra ns faithful re the ru ru ma ma to the ACP for the lonm-A CP weapon. Then, a reaction of condemnation is produced, where the majority of the CP went only with the acetyl-C or A, giving com or result or the power to ac il-A CP. The p-ce to ac il-A CP s in ta sa III (F ab H) in ic the FAS code, after the p-ce to ac il-A CP s in ta sa I ( F ab B) and the p-ce to ac il-A CP s in ta sa II (F abF) is as implied in the possible positions.

A co n tin u a c io n , se rep ite un c ic lo de e ta p a s ha s ta o b te n e r un a c id o g ra so sa tu ra d o de lo ng itud a p rop iad a . En p rim er lugar, la N A D P H reduce la p -ce to a c il-A C P para fo rm a r p -h id ro x ia c il-A C P . Esta e tapa la ca ta liza la p -ce to a c il-A C P red u c ta sa (FabG ). D espues, la p -h id ro x ia c il-A C P se de sh id ra ta para fo rm a r tra n s -2 -e n o il-A C P . La p -h id ro x ia c il-A C P d e s h id ra ta s a /is o m e ra s a (F a b A ) o la p -h id ro x ia c il-A C P d e s h id ra ta sa (F abZ ) ca ta liza n esta e tapa . La tra n s -2 -e n o ilA C P re d u c ta sa I, II o III (F ab I, FabK , o FabL , re sp e c tiva m e n te ) d e p e n d ie n te de N A D P H , red uce la tra n s -2 -e n o il-A C P para fo rm a r ac il-A C P . Los c ic los po s te rio res se in ic ian a tra ve s de la co n d e n sa c io n de m a lon il-A C P con ac il-A C P m ed ia n te la p -ce to a c il-A C P s in ta sa I o la p -ce to a c il-A C P s in ta sa II (F abB o FabF, resp ec tiva m en te ).

In accordance with this, a cycle of this country has been repeated so that it is possible to obtain your clothing. First, NADPH reduces the p-ce to ac il-A CP for fo rm arp -h id ro x ia c il-A CP. This covers the p-ce to ac il-A CP red uc ta sa (FabG). Then, the p-h id ro x ia c il-A CP is sh id ra ta for fo rm ar tra ns -2 -eno il-A CP. The p-h id ro x ia c il-A CP desh id ra ta sa / is ome ra sa (F ab A) or the p-h id ro x ia c il-A CP desh id ra ta sa (F abZ ) This is the cover. Tra ns -2 -eno ilA CP re duc ta sa I, II or III (F ab I, FabK, or FabL, re sp e c tive ment) depending on NADPH, red it translates ns -2 -eno il-A CP for fo rm ar ac il-A CP. The powers are initiated through the longest-to-first CP with the highest number of members. -A CP s in ta sa I or the p-ce to ac il-A CP s in ta sa II (F abB or FabF, resp. M in te).

II. M od ifica c io n de la ru ta b io s in te tica de ac ido s g ra sos para a u m e n ta r la p ro du cc ion de ac il-A C P

II. M od ifica tio n the route in ac tica de ac a tica s g ra sos for a u m e n ta r la p ro du c ation of ac il-A C P

Los o rg a n ism o s ho sp e d a d o re s de p ro du cc ion pueden g e n o m a n ip u la rse para s o b re p ro d u c ir a ce til-C o A y m a lo n il-C oA . D ichos o rg a n ism o s ho sp e d a d o re s de p ro du cc ion inc luyen ce lu las ve g e ta le s , an im a les o hum anas. C om o h o sp e d a d o re s de p roducc ion pueden u tiliza rse m ic ro o rg a n ism o s ta le s com o c ia n o b a c te ria s , a lgas, bacterias, le va d u ra s u hongos fila m en to sos . C om o e je m p lo s no lim ita tivos de m ic ro o rg a n ism o s que pueden u tiliza rse com o h o sp e d a d o re s de p ro du cc ion se in c luyen E. coli, Saccharomyces cerevisiae, Candida lipolytica, Synechococcus, Synechocystis, Clamydomonas, Arthrobacter AK 19, Rhodotorula glutinins, Acinetobacter sp. cepa M-1, Candida lipolytica, y o tros m ic ro o rg a n ism o s o lea g in oso s . P ara o b te n e r una m a y o r p ro du cc ion de a ce til-C o A /m a lo n il-C o A /a c id o s g ra sos y d e riva d o s de ac ido s g rasos, se pueden h a ce r va r ia s m od ifica c io n e s d ife ren te s , ya sea en co m b in a c io n o in d iv idu a lm e n te , en el h o s p e d a d o r de p roducc ion .

Those who have been given the right to be able to do so can go to go to C-C or A and even in C-A. Sayings or rg an ism os ho sp edado re s de p ro du cc ion inc lu ve c ve lu ve le ta ta s, an im ales or hu annas. As long as possible, they can be used by themselves or not only as long as they are, however, bacteria, their fungi are going to be in line. . For example, it is not possible to limit the benefits that can be used as a basis for the protection of E. coli, Saccharomyces cerevisiae, Candida lipolytica , Synechococcus, Synechocystis, Clamydomonas, Arthrobacter AK 19, Rhodotorula glutinins, Acinetobacter sp. strain M-1, Candida lipolytica, yo tros m ic ro or rg an ism oso lea g in oso s. In order to obtain a greater degree of interest in C-A or A / m at least C or A / C id. r ia sm od ifica c io nesd ife ren te s, whether in co mb in ac io not in d iv idu a lm in te, in the host of production.

P o r e jem p lo , para a u m e n ta r la p ro d u cc io n de ace til-C o A , en un h o sp e d a d o r de p ro du cc ion puede exp resa rse uno o m as de los s ig u ie n te s genes: pdh, panK, aceEF (que cod ifica el co m p o n e n te E 1p d e s h id ro g e n a sa y el com p o n e n te E 2p d ih id ro lip o a m id a a c iltra n s fe ra sa de los com p le jos de p iruva to y 2 -o xo g lu ta ra to de sh id ro g e n a sa ), fabH, fabD, fabG, acpP y fabF. En o tros e jem p los , en el h o s p e d a d o r de p ro du cc ion pueden exp re sa rse ge ne s a d ic io n a le s que co d ifica n a c il-C o A re d u c ta sa s g ra sas y a ld e h id o de sca rb o x ila sa s . En la m a te ria se sabe que puede co n s tru irse un p lasm id o que con te n g a uno o m as de los genes m e n c io n a d o s a n te rio rm en te , to d o s e llos ba jo el con tro l de un p ro m o to r co n s titu tivo , o de o tro m odo, con tro la b le . En los p a re n te s is se in d ica n los n u m e ro s de reg is tro de G e n B a n k e je m p la re s de estos genes: pdh (B A B 34380 , A A C 73227 , A A C 73226 ), panK ( tam b ien con oc ido com o coaA, A A C 76952 ), aceEF (A A C 73227 , A A C 73226 ), fabH (A A C 74175 ), fabD (A A C 74176 ), fabG (A A C 74177 ), acpP (A A C 74178 ) y fab F (A A C 74179 ).

P o jem p lo, in order to increase the pro duction of acetyl-C or A, one or more of the six years may expire The genes are: pdh, panK, aceEF (which codifies the co mpone te E 1p desh id ro gena sa and the com put te E 2p d ih id ro lip oam id aac iltra ns fe ra sa of the com p le jos from p iruva to y 2 -o xo g lu ta ra to from sh id ro gena sa), fabH, fabD, fabG, acpP and fabF. In other countries, in the host of the pipeline, it can be expressed that it is known as a national or C or A duc ta sa sg ra sas ya ld eh id of sca rb ox ila sa s. In the morning, it is known that there can be a long way to go with one or more of the genes mentioned in the world, taking them under the control of a person By way of constituent, or in a way, with the bank. In the countries of the registers of G in B anke je mp the re of these genes: pdh (BAB 34380, AAC 73227, AAC 73226), panK (also with a common agreement, AAC 76952), aceEF (AAC 73227, AAC 73226), fabH (AAC 74175), fabD (AAC 74176), fabG (AAC 74177), acpP (AAC 74178) and fab F (AAC 74179).

A d ic io n a lm e n te , los n ive les de exp res ion de fadE, gpsA, ldhA, pflb, adhE, pta, poxB, ackA y /o ackB, pueden a te n u a rse o in ac tiva rse en el m ic ro o rg a n ism o g e n o m a n ip u la d o m ed ia n te tra n s fo rm a c io n con p lasm idos co n d ic io n a lm e n te re p lica tivo s o no re p lica tivo s que con tienen m u ta c io n e s nu las o po r de le c ion de los genes c o rre sp o n d ie n te s , o su s titu ye n d o las s e cu e n c ia s p ro m o to ra s o p o tenc iad o ras . En los p a ren te s is se ind ican los nu m ero s de reg is tro de G e n B a n k e je m p la re s de estos genes: fadE (A A C 73325 ), gspA (A A C 76632 ), ldhA (A A C 74462 ), pflb (A A C 73989 ), adhE (A A C 74323 ), pta (A A C 75357 ), poxB (A A C 73958 ), ackA (A A C 75356 ) y ackB (B A B 81430 ). Los ho sp e d a d o re s de p ro du cc ion g e n o m a n ip u la d o s re su lta n te s tie n e n n ive les de p ro du cc ion de ace til-C o A au m e n ta d o s cua nd o se cu ltivan en un e n to rno ap rop iado .

On a daily basis, the export levels of fadE, gpsA, ldhA, pflb, adhE, pta, poxB, ackA and / or ackB, can be added or activated in the My ic o rg an ism ogenoman ip u la dom ed ia n te tra ns fo rm ac io n with plasmids co nd ic io na lm in te p r tica tivo so no re p lica tivo s they have mu This is also the case of the genes of the common genes, or their location is the same as the tenor. In the countries the registration numbers of G are indicated in B anke je mp the re of these genes: fadE (AAC 73325), gspA (AAC 76632), ldhA (AAC 74462) , pflb (AAC 73989), adhE (AAC 74323), pta (AAC 75357), poxB (AAC 73958), ackA (AAC 75356) and ackB (BAB 81430). The most recent employees of Genoman IP are the two most important in the case of acetyl-C or A two-month increase. in an en ap rno ap rop iado.

A d e m a s , la so b re p ro d u cc io n de m a lo n il-C o A puede ve rse a fe c ta d a g e n o m a n ip u la n d o el h o s p e d a d o r de p roducc ion co m o se d e sc rib e a n te rio rm e n te con accABCD (p o r e jem p lo , nu m ero de reg is tro A A C 73296 de G en B ank, EC 6.4.1.2 ) in c lu ido en el p lasm id o s in te tiza d o de novo. La s o b re p ro d u cc io n de ac ido s g ra sos puede rea liza rse in c luye nd o a d ic io n a lm e n te un gen que cod ifica la lipasa (p o r e jem p lo , num eros de reg is tro C A A 89087 y C A A 98876 de G e n B a n k ) en el p la sm id o s in te tiza d o de novo.

In addition, the underwriting of ma n-C or A can be seen to date in which the host of production as well as sc rib ean te rio rm in te with accABCD (pore jem p lo, registration number AAC 73296 de G en B ank, EC 6.4.1.2) in c p gone in the pasm id os in te tiza do de de de novo. The surplus of the accent of the following can be carried out in a gene that encodes the lipase (pore jem p lo, numbers of reg. Tro CAA). 89087 and CAA 98876 of G in B ank) in the p la sm id os in te tiza do de de novo.

C o m o resu ltad o , en a lg u n o s e jem p los , una a c e til-C o A ca rb o x ila sa se s o b re e x p re s a pa ra a u m e n ta r su c o n ce n tra c io n in tra ce lu la r en al m en os a p ro x im a d a m e n te 2 vece s , al m en os a p ro x im a d a m e n te 5 vece s o al m enos a p ro x im a d a m e n te 10 vece s , con resp ec to a los n ive les de exp re s io n na tivos.

As a result, in some cases, an acetyl-C or A ca rb ox ila sa is overdue to increase its con- traction with the intra-ce r at al m in os ap ro x im adamen te 2 times s, at m in os ap ro x im adamen te 5 times so at m apos ap ro x im adamen te 10 times s, with respect to the levels They are native speakers.

A d e m a s , para a u m e n ta r la can tida d de a c il-C o A d ispon ib le , puede u tiliza rse un m u tan te D 311 E P lsB (p o r e jem p lo , nu m ero de reg is tro A A C 77011 de G enB ank).

In addition, to increase the amount of ac-C or A d ispon ib le, a person can be used as D 311 EP lsB (for example, registration number AAC 77011 of G enB ank).

A d e m a s , la so b re e xp re s io n de un gen sfa (s u p re s o r de FabA , po r e jem p lo , nu m ero de reg is tro A A N 79592 de G e n B a n k ) puede in c lu irse en el h o sp e d a d o r de p ro du cc ion para a u m e n ta r la p ro du cc ion de ac idos grasos m o n o in sa tu ra d o s (R o cke t al., J. B ac te rio logy , 178 :5382 -5387 , 1996).

In addition, the so b re e xp re s io n of a sfa gene (supra de fab sor, po re jem p lo, registration number AAN 79592 of G in B ank) can in c Go to the old age of P ro du cc ion to increase the p r du du rion of fatty acids mono in sa tu ra two (R o cke t al., J. B ac te rio logy, 178 : 5382-5387, 1996).

B. Acil-ACP y/o Acil-CoA a ester graso utilizando tioesterasa

B. Acyl-ACP and / or Acyl-CoA to fatty ester using thioesterase

En un m od e lo tip ico de p ro ceso m ic ro b ia n o de s in tes is de a c id o s g rasos, la a c e til-C o A y la m a lo n il-C o A se tra n s fo rm a n a tra ve s de una se rie de e ta pas para fo rm a r las cad en as de ac il-A C P . despues , la A c il-A C P se tra n s fo rm a , m ed ia n te una se rie de e ta pas en z im a tica s a lte rna tivas , a d ive rso s p ro du c to s fina les , in c luye nd o d e riva d o s de ac ido s grasos. P o r e jem p lo , no rm a lm e n te , la a c il-A C P se tra n s fo rm a en e s te re s g ra sos m ed ia n te las re a cc io n e s co n se cu tiva s co m b in a d a s de una tio e s te ra sa , una a c il-C o A lig a sa /s in te ta sa y una e s te r s in tasa . Una lim itac io n al uso com e rc ia l de es ta s e n z im a s en una ru ta m e ta b o lica es la ne ces idad de p ro d u c ir el sus tra to de acil C o A g ra so a p a rtir de un p re cu rso r de ac il A C P graso , que requ ie re al m e n o s dos e ta pas e n z im a tica s y el gasto de e n e rg ia m e ta b o lica de dos en laces fo s fo a n h id rid o s . La p roducc ion d irec ta de es te res g ra sos con t io e s te ra sa m itiga la pe rd ida de A T P ca u sa d a po r es ta s dos e ta pas en z im a ticas . R e c ie n te m e n te se ha d e m o s tra d o que las lipasas (cuyo sus tra to de "a lcoh o l" na tu ra l es el ag ua ) ta m b ie n pueden u tiliza rse in vitro pa ra c a ta liza r la reacc ion de tra n s e s te r if ic a c io n que p ro d u ce el b iod iese l (es decir, la tra n s fo rm a c io n de tr ia c il g lice rid o y m e tan o l en e s te r m e tilico de ac ido g raso y g lice ro l). S in em bargo , las lipasa s son g e n e ra lm e n te to x ica s en las ce lu la s cua nd o se p roducen in tra ce lu la rm en te .

In a typical way of doing so, it is not a matter of symbols, the ace-C or A and the most C-A or A are For the purpose of a laugh at this country, for the purposes of ac-A-CP. afterwards, the A c il-A CP will be carried out, by means of a laugh at this point of interest, ad ive rso sp ro du c to s fina les, In c luye nd ode riva two of fatty acids. P o jem p lo, not rm a lm in te, the ac il-A CP will be the only one in which you will be re g ected in re n a cions Co mb in adas un un es es ra ra, an ac il-C or A lig a sa / s in te ta sa and one e s te rs in rate. A limitation to the common use of this sen- sity in a basic route is the need for money to go through the accrual treatment C o A g ra so apa It is the first step of the ACP fatty, which requires at least two enzymatic countries and the expenditure of two basic medical expenses in the foor fo anh fo ridicul. you The direct production of these rights with you is the reason why ATP's loss falls within two months. R ec ie n it has been shown to you that lipases (whose treatment of "a lcoh ol" na tu ra l is ag ua) can also be used in vitro for the purpose of liza r the reaction of tra nses te r if ic ac io n that p ro du ce the b iod iese l (that is, the tra ns fo rm a tio of tr ia c il g lice rid oyme so ol in that s I love you, lime de ac gone g sato yg lice ro l). However, the lipases are generally in toxic terms in which they are produced in rm in te rm.

A p e sa r de te n e r una e sp e c ific id a d p u b licad a po r el agua , la p re se n te d ivu lg a c io n d e sc rib e el d e scu b rim ie n to de que, en p re sen c ia de una can tidad su fic ie n te de a lcoho l, el a lcoh o l puede lle g a r a s e r un sus tra to a ce p tab le para una tio e s te ra sa . En este caso , las t io e s te ra sa s pueden c a ta liz a r la a lcoh o lis is de los p ro du c to s in te rm e d io s de las en z im a s de ac ilo graso , al igua l que hace una lipasa in vitro. P or tan to , en las co n d ic io n e s ad ecu ad as , puede u tiliza rse una en z im a que ace p te un e s te r g ra so com o sus tra to para fo rm a r un p ro du c to in te rm e d ia rio de en z im a g ra sa que po s te rio rm e n te se esc inda po r h id ro lis is o tra n se s te r if ica c io n para s in te tiz a r es te res de ac ido s grasos d e s e a d o s si se p ro p o rc io n a un n ive l s u fic ie n te de un a lcoh o l a d e cu a d o pa ra a c tiv a r la a lcoh o lis is . C o m o e jem p los de en z im a s que tien en esta cap ac idad , que pueden p ro d u c ir e s te re s d ire c ta m e n te a p a rtir de ac il-A C P se inc luyen , ad e m a s de tio e s te ra sa s , a c iltran s fe rasas , lipasas, es te ra sas y p ro teasas. Las t io e s te ra sa s u tiles pueden se r t io e s te ra sa s na tu ra les y/o p re cu rso ra s com o se de fine en el p resen te d o cum e n to , o pueden s e r tio e s te ra sa s m u tan tes p re pa rada s segun se de sve la en el p re sen te do cum e n to . Un exp erto hab itua l en la m a te ria es capaz de d e te rm in a r la co n ve n ie n c ia de u tiliza r una e n z im a p a rticu la r pa ra p ro d u c ir d ire c ta m e n te e s te re s g ra sos a p a rtir de A c il-A C P . P o r e jem p lo , los e n sa yo s p ro p o rc io n a d o s en el E je m p lo 32 son u tiles pa ra d e te rm in a r la p ro du cc ion d irec ta de este r.

In spite of having a special idea for water, the reason why the program is that of scu b rim ie n that In the sense of a quantity that is enough for him, the spirit may be able to make a deal with him for an uncle. In this case, the types that can be used for the purposes of the fatty acids in the fatty acid areas, at the same time l that makes a lipase in vitro. Therefore, in the ad hoc regions, one can be used in this way so that one can only be treated as his or her treats to promote a certain period of time. c to in te rm ed ia rio de en z im ag ra sa that po s te rio rm en te es in inda po rh id ro lis is o tra n se r te ica c io n for s in te tiz ar These are the desired fatty acids if a single level of income is allowed for the purpose of acquiring the age. How do you think that you have in this capacity, that you can go to see if you are going to leave this area? Uncle is te ra sa s, ac iltran s fe rasas, lipasas, es te ra sas yp ro teasas. Those who are useful are those who are aware of their lives and / or how they are fine in the country, or they may be uncle It is te sa sauu so tes p re pa rada s as per sve la in the p re sen te cum en to. An ordinary resident in the ma- ria is able to tell you how to use an enzymatic device to carry out the duo go to the direction of the men This is what you need to say in A c il-A CP. P o jem p lo, those in sa me sp ro po rc io swim in the E je mp lo 32 are useful for the r direct in the direct direction of this r r.

S egun este asp e c to de la d ivu lg ac ion , la t io e s te ra sa puede u tiliza rse para p ro d u c ir d ire c ta m e n te e s te re s g ra sos en p re se n c ia o en au se n c ia de una e s te r s in ta sa y /o una ac il C o A lig a sa /s in te ta sa grasa . P o r e jem p lo , la exp res ion de una t io e s te ra s a que puede c a ta liz a r la p ro du cc ion d irec ta de es te res g ra so s en una cepa ho spe d a d o ra re co m b in a n te , pued e u tiliza rse pa ra c o m p le m e n ta r la p ro d u cc io n de e s te r g ra so do n d e la cep a ta m b ie n exp re sa una e s te r s in tasa . A d ic io n a lm e n te , la exp res ion de una t io e s te ra s a que puede c a ta liz a r la p ro du cc ion d irec ta de este res g ra sos en una ce lu la h o sp e d a d o ra reco m b in an te , puede u tiliza rse dond e no hay exp res ion de e s te r s in ta sa o esta e xp res ion es baja.

According to this aim of the decision, the uncle can be used to go to the direction of his life. In this case, you will be in and / or a joint venture will be fat. For the sake of an uncle's expression, it is possible that he can determine the direction of the latter in a household strain given by him or her. In addition, you can use it to make sure that you are happy with the idea that you will be at a rate of interest. On a daily basis, the expiration of an uncle is that it may be possible to direct the direct debit of the latter in a household. In this case, it can be used where there is no expiration of this one or this is low.

P uede u tiliza rse una t io e s te ra sa m u tan te que se ha m od ifica do para que te n g a p ro p ieda de s a lte ra d a s en c o m p a ra c io n con la t io e s te ra s a p recu rso ra .

It is possible for you to become an uncle, it is very much so that you have been measured so that you will never be able to compare yourself with the uncle. ra.

C. Acil-ACP a acido graso

C. Acyl-ACP to fatty acid

I. R u ta b ios in te tica de ac ido s g rasos: a c il-A C P a ac ido s g rasos

I. R u ta b ios in the art of ac i s satin: a c il-A C P a ac i s s satin

T a l com o se ha de sc rito an te rio rm e n te , la a c e til-C o A y la m a lo n il-C o A se p rocesan en va ria s e ta pas para fo rm a r ca d e n a s de ac il-A C P . La e n z im a sn -g lice ro l-3 -fo s fa to a c iltra n s fe ra sa (P lsB ) ca ta liza la tra n s fe re n c ia de un grupo ac ilo de ac il-A C P o a c il-C o A a la po s ic io n sn-1 de l g lice ro l-3 -fo s fa to . P o r tan to , la P lsB es una e n z im a reg u lado ra c la ve en la s in tes is de fos fo lip ido s , que fo rm a de la ru ta de los ac ido s g rasos. La inh ib ic ion de P lsB con du ce a un au m e n to en los n ive les de ac il-A C P de cadena larga, cuya re tro a lim e n ta c io n in h ib ira e ta pas te m p ra n a s en la ruta, que im p lica genes ta le s com o , po r e jem p lo , accABCD, fabHy fabl. El d e sa co p la m ie n to de esta reg u lac io n , por e jem p lo , po r so b re e xp re s io n de tio e s te ra sa , co n d u ce a una m a yo r p ro du cc ion de ac ido s grasos.

As usual, r r r e rm in te, acetyl-C or A and most of the C-A are in contact with each other for the purpose of accruals. il-A CP. The enzyme im sn -g lice ro l-3 -fo s fa to ac iltra ns fe ra sa (P lsB) can be used as the basis of an acyl group of ac il-A CP oac il -C or A at the sn sn-1 of lg lice ro l-3 -fo s fa to. Therefore, the P lsB is a regular agency that sees it in the symposiums of phosphorus, which is the path of the future events. The introduction of P lsB with an increase in the levels of long-chain CPs, whose return is limited in h ib will go to the country. On the route, which involves genes that are common, such as accABCD, fabHy fabl. The one in this region, for example, because it is uncle's name, it is known to a mother ro du cc ion of fatty acids.

II. M od ifica c ion de la ruta b ios in te tica de ac ido s g rasos para p ro d u c ir los tip o s d e sea do s o las p ro po rc ion es d e se a d a s de ac ido s g ra sos

II. M od ification of the route b ios in te tica de ac a s gs satos para p ro d u c go the types of the two or the p o po r r ion ion d e se a d a s a d a s de ac a s g ra sos

S egun la p re sen te d ivu lg ac ion , la tio e s te ra s a e xp resa da tien e p ro p ie d a d e s a lte rad as en com p a ra c io n con la t io e s te ra s a na tiva o e n d o ge na en la cepa ho spe da dora . P a ra d is e n a r un h o s p e d a d o r de p ro du cc ion para la p ro du cc ion de una po b lac ion h o m og ene a de de riva dos de ac ido s grasos, uno o m as genes en d o g e n o s pueden a te n u a rse o de le c ion a rse fu n c io n a lm e n te y, com o resu ltado , pueden e xp re sa rse una o m as t io e s te ra sa s segun la d ivu lg ac ion . P o r e jem p lo , a te n u a n d o una tio e s te ra s a C ¹⁸ (p o r e jem p lo , nu m ero s de reg is tro A A C 73596 y P 0A D A 1 de G en B ank), que u tiliza C ¹⁸ : ¹-A C P , y exp re sa n d o una tio e s te ra s a a lte rada que tien e una m a y o r e sp ec ific ida d po r y/o ac tiv ida d (p o r e jem p lo , v e lo c id a d ca ta litica ) con re sp e c to a sus tra to s C ¹⁰ (es decir, sus tra to s que com p ren den , cada uno de e llos, una long itud de 10 ca rb o n o s en la cad en a de ca rb on os), pu ed en p ro du c irse de riva dos de ac ido s g ra sos C ¹⁰ (es decir, d e riva d o s de ac ido s g ra sos que com p ren den , cada uno de e llos, una long itud de 10 ca rb o n o s en la cad en a de ca rb on os). Esto da com o resu ltad o una po b lac ion m as ho m o g e n e a de de riva d o s de ac ido s g rasos que tie n e n un a u m en to en ac idos g ra sos que tien en una lo ng itud de 10 ca rb on os en la cadena . En otro e jem plo , a te n u a n d o tio e s te ra s a s en d o g e n a s que p roducen ac ido s g ra sos n o -C ¹² y exp re sa n d o una tio e s te ra sa a lte rad a que tie n e una m a yo r e sp ec ific ida d po r y /o ac tiv idad (es decir, v e lo c id a d ca ta litica ) con resp ec to a su s tra to s C ¹² , pueden p ro du c irse de riva dos de ac ido s g rasos C ¹². En otro e jem p lo , a te n u a n d o t io e s te ra sa s en d o g e n a s que p roducen a c id o s g ra so s no C ¹⁴ y e x p re s a n d o una tio e s te ra s a a lte ra d a que te n g a una m a y o r e sp e c ific id a d po r y /o ac tiv id a d (es decir, v e lo c id a d ca ta litica ) con resp ec to a sus tra to s C ¹⁴ , pueden p ro du c irse d e riva d o s de ac ido s g ra sos C ¹⁴. En otro e jem p lo , en la m ezc la de l p ro du c to puede o b tene rse un re n d im ie n to p ro p o rc io n a l m as a lto de d e riva d o s de ac idos g ra so s de ca d e n a co rta (p o r e jem p lo , C8, Cg, C ¹⁰ , C ¹¹ , C ¹² , C ¹³ y /o C ¹⁴ ) fre n te a o tros d e riva d o s de a c id o s g ra so s de cad en a no corta. En otro e jem p lo m as, en la m ezc la del p ro du c to ta m b ie n puede o b tene rse un ren d im ien to p ro p o rc io n a l m as ba jo de de riva dos de ac ido s g ra sos de cadena co rta (p o r e jem p lo , C8, Cg, C ¹⁰ , C ¹¹ , C ¹² , C ¹³ y/o C ¹⁴ ) fren te a o tros de riva dos de ac ido s g ra sos no de cadena corta . En un e jem p lo a d ic io na l, exp re sa n d o una t io e s te ra s a a lte ra d a que te n g a ve lo c id a d ca ta litica y /o p ro d u cc io n o re n d im ie n to m e jo rad os in vivo, puede p ro du c irse un rendimiento mas alto y/o mejorado de derivados de acidos grasos libres. En otro ejemplo mas, aplicando una o mas de ciertas mutantes de tioesterasa puede producirse un rendimiento proporcional o porcentual de esteres grasos mas alto o mas bajo frente a otros productos, tales como acidos grasos libres. La sobreproduccion de acetil-CoA, malonil-CoA y acidos grasos puede verificarse utilizando metodos conocidos en la materia, por ejemplo, por precursores radiactivos, HPLC, LC-Ms y GC-MS posteriores a la lisis celular.

According to the p re sen te d ivu lg ac ion, the uncle is te sa sa xp resa da tien ep ro p ie dadesa lte rad as in comparison with the un io is te ra sa na tiva oend o Gen na in the strain spe spe da dora. P a ra d is e n ar a host of p ro du cc ion for the p ro du cc ion of a hom og e a r ive of two fatty acids, one or more genes in d o genes can be added or read to go fu nc io na lm en te and, as a result, they can be an other person according to their decision . P o jem p lo, when an uncle is tera sa C ¹⁸ (for example, reg. Number AAC 73596 and P 0A DA 1 of G in B ank), which uses C ¹⁸ : ¹ -ACP, and exp re sa ndo an uncle is te ra saa lte rada who has a major sp ec ific ida d po ry / o ac tiv ida d (pore jem p lo, see c id ad ca ta litica ) with respect to their C ^{10 treatments} (that is, their contracts that comprise, each of them, a length of 10 years in the cadre of each of them) os), in order to leave from Riva Dos de Ac Ido Sg Ra SOS ¹⁰ (that is, from Riva Dos de Ac Ido Sg Ra, so that each one of them may have a long history) of 10 cars in the city of ca). This results in a more or more recent population of two countries that have an increase in the number of acidic acids they have in a 10-year period. rb on os in the chain. In another example, when you are an uncle, you are in days who are still ac- cessed if you are not -C ¹² and an uncle is exposed to the rad radius of a mother I re sp ec ific ida d po ry / o ac tiv idad (that is, see the id ad ca ta lithic) with respect to its system C ¹² , can p ro du c leave riva two of Goed in C ¹² . In another jem p lo, a nuandot io is te ra sa s in d o genas that p rduced a c id osg ra so s no C ¹⁴ yexp re sando uncle es te ra saa lte ra da nga a majore sp ec ific id ad po ry / o ac tiv id ad (that is, see what id id ca l lithic) with respect to their work C ¹⁴ , they can leave riva two of acido sg ra sos C ¹⁴ . In another case, in the case of the pipeline, a net income can be obtained at the end of two years of acidic acids. cana dena co rta (pore jem p lo, C8, Cg, C ¹⁰ , C ¹¹ , C ¹² , C ¹³ and / or C ¹⁴ ) fre n te ao tros de riva dos de a c id osg ra so s de cad en to not cut. In another case, at most, in the case of the government, there can also be a renouncement to the end of the year. sg ra sos de co rta chain (pore jem p lo, C8, Cg, C ¹⁰ , C ¹¹ , C ¹² , C ¹³ and / or C ¹⁴ ) fren te ao tros de riva dos de acc sg ra sos not a chain short . In a je p p ad na io na l, exp re sa ndo un un es es te ra saa lte ra da nga see what the id id ca ca lithic y / op ro du cc io no re nd im ie n I can go live, you can leave a higher and / or improved yield of free fatty acid derivatives. In another example, by applying one or more of certain thioesterase mutants, a proportional or percentage yield of higher or lower fatty esters can be produced compared to other products, such as free fatty acids. The overproduction of acetyl-CoA, malonyl-CoA and fatty acids can be verified using methods known in the art, for example, by radioactive precursors, HPLC, LC-Ms and GC-MS subsequent to cell lysis.

En un aspecto alternativo, una tioesterasa de la divulgacion puede expresarse dentro de la cepa hospedadora en combinacion con una tioesterasa endogena. En otro aspecto alternativo, una o mas tioesterasas endogenas pueden modificarse utilizando tecnicas de alteracion genomica adecuadas que son conocidas por los expertos en la materia, de modo que las tioesterasas mutantes tengan al menos una propiedad alterada en comparacion con los precursores de tioesterasa endogenos, y/o de tal manera que la celula hospedadora presente al menos una propiedad alterada en comparacion con la celula hospedadora antes de que se apliquen dichas tecnicas de alteracion genomica.

In an alternative aspect, a thioesterase of the disclosure can be expressed within the host strain in combination with an endogenous thioesterase. In another alternative aspect, one or more endogenous thioesterase can be modified using suitable genomic alteration techniques that are known to those skilled in the art, so that the mutant thiosterases have at least one altered property compared to the endogenous thioesterase precursors, and / or in such a way that the host cell has at least one altered property compared to the host cell before such genomic alteration techniques are applied.

D. Acidos grasos en Acil-CoA

D. Fatty acids in Acil-CoA

I. Transformacion de acidos grasos en acil-CoA

I. Transformation of fatty acids in acyl-CoA

La acil-CoA sintasa (ACS) esterifica acidos grasos libres a acil-CoA mediante un mecanismo de dos etapas. En primer lugar, el acido graso libre se transforma en un producto intermedio de acil-AMP (un adenilato) a traves de la pirofosforolisis de ATP. Despues, el carbono carbonilo activado del adenilato se acopla al grupo tiol de CoA, liberando AMP y el producto final acil-CoA. Vease Shockey et al., Plant Physiol. 129:1710-1722, 2002.

Acyl-CoA synthase (ACS) esterifies free fatty acids to acyl-CoA through a two-stage mechanism. First, free fatty acid is transformed into an intermediate product of acyl-AMP (an adenylate) through the pyrophosphorolysis of ATP. Then, the activated carbonyl carbon of the adenylate is coupled to the thiol group of CoA, releasing AMP and the final acyl-CoA product. See Shockey et al., Plant Physiol. 129: 1710-1722, 2002.

La enzima ACS FadD de E. coli y la proteina de transporte de acidos grasos Fadl normalmente son componentes importantes de un sistema de absorcion de acidos grasos. Fadl actua como mediador en el transporte de acidos grasos en la celula bacteriana, y FadD actua como mediador en la formacion de esteres de acil-CoA. Cuando no se dispone de otra fuente de carbono, las bacterias absorben los acidos grasos exogenos y se transforman en esteres de acil-CoA, que se unen al factor de transcripcion FadR y desreprimen la expresion de los genes fad que codifican proteinas responsables del transporte (Fadl), la activacion (FadD) y la p-oxidacion (FadA, FadB, FadE y FadH) de los acidos grasos. Cuando se dispone de fuentes de carbono alternativas, las bacterias sintetizan acidos grasos tales como las acil-ACP, que despues se utilizan para la sintesis de fosfolipidos, en lugar de servir como sustratos para la p-oxidacion. Por tanto, la acyl-CoA y la acyl-ACP son fuentes independientes de acidos grasos que conducen a diferentes productos finales. Vease Caviglia et al., J. Biol. Chem., 279(12):1163-1169, 2004.

The ACS FadD enzyme from E. coli and the Fadl fatty acid transport protein are usually important components of a fatty acid absorption system. Fadl acts as a mediator in the transport of fatty acids in the bacterial cell, and FadD acts as a mediator in the formation of acyl-CoA esters. When no other carbon source is available, bacteria absorb exogenous fatty acids and transform into acyl-CoA esters, which bind to the FadR transcription factor and suppress the expression of fad genes that encode proteins responsible for transport ( Fadl), activation (FadD) and p-oxidation (FadA, FadB, FadE and FadH) of fatty acids. When alternative carbon sources are available, bacteria synthesize fatty acids such as acyl-ACP, which are then used for phospholipid synthesis, instead of serving as substrates for p-oxidation. Therefore, acyl-CoA and acyl-ACP are independent sources of fatty acids that lead to different end products. See Caviglia et al., J. Biol. Chem., 279 (12): 1163-1169, 2004.

II. Modificacion de la ruta biosintetica de acidos grasos para aumentar la transformacion de acidos grasos a acil-CoA

II. Modification of the biosynthetic route of fatty acids to increase the transformation of fatty acids to acyl-CoA

Los hospedadores de produccion pueden genomanipularse utilizando peptidos que se sabe que producen acidos grasos de diversas longitudes que pueden transformarse en acil-CoA. Un metodo para producir derivados de acidos grasos implica aumentar la expresion de, o expresar formas mas activas de, uno o mas peptidos de acil-CoA sintasa (EC 6.2.1.-).

Production hosts can be genomanipulated using peptides that are known to produce fatty acids of various lengths that can be transformed into acyl-CoA. A method for producing fatty acid derivatives involves increasing the expression of, or expressing more active forms of, one or more acyl-CoA synthase peptides (EC 6.2.1.-).

En la Tabla 1 se muestra una lista de acil-CoA sintasas que pueden expresarse para producir acil-CoA y derivados de acidos grasos. Estas acil-CoA sintasas pueden examinarse para optimizar cualquier ruta que utilice acil-CoA grasas como sustratos. Utilizando tecnicas de bioinformatica y genes sinteticos, pueden expresarse genes fadD heterologos en cepas de produccion y evaluar su capacidad para producir biodiesel y posiblemente biocrudo.

Table 1 shows a list of acyl-CoA synthases that can be expressed to produce acyl-CoA and fatty acid derivatives. These acyl-CoA synthases can be examined to optimize any route that uses acyl-CoA fats as substrates. Using bioinformatics techniques and synthetic genes, heterologous fadD genes can be expressed in production strains and assess their ability to produce biodiesel and possibly biocrude.

Tabla 1: Acil-CoA sintasas

Table 1: Acyl-CoA synthases

co n tin u a c io n

continuation

En fun c ion de su g rado de s im ilitud con fadD de E. coli, se s e le cc io n a ro n los s ig u ie n te s genes h o m o logo s para s in te tiza rse y eva lua rse :

Based on its degree of similitude with fadD of E. coli, the homologous genes will be used to identify and evaluate it. :

fadDD35 de M. tuberculosis H R 7 R v [N P _217021 ].

fadDD35 of M. tuberculosis HR 7 R v [NP_217021].

yh fL de B. subtilis [N P _388908 ].

and h fL of B. subtilis [NP_388908].

fadD1 de P. aeruginosa PAO1 [N P _251989 ].

fadD1 of P. aeruginosa PAO1 [NP _251989].

h o m o logo de fadD, que cod ifica Faa3p de Saccharomyces cerevisiae [N P _012257 ].

hom or fadD logo , which encodes Faa3p of Saccharomyces cerevisiae [NP _012257].

C o m o a c il-C o A s in ta sa s de ac ido s g ra sos a d ic io n a le s de o rg a n ism o s euca rio tas , que pu ed en u tiliza rse para p ro d u c ir ac il-C oA , a s i com o de riva dos de ac idos g rasos, se inc luyen las d e sc rita s en S h o cke y e t al., P la n t P hys io l., 129: 1710 -1722 , 2002 (Arabidopsis), C a v ig lia e t al., J. B iol. C hem ., 279: 1163 -1169 , 2004 (ra ta ) y en K no ll e t al., J. Biol. C hem ., 269(23): 16348-56 , 1994 (le vad u ra ). En la te cn ica se conocen se cu e n c ia s ge n icas que cod ifican estas s in te tasas . V ease , po r e jem p lo , Jo h n so n e t al., J. B io l. C hem ., 269: 18037 -18046 , 1994; S h o cke y e t al., P lan t P hys io l., 129: 1710 -1722, 2002 ; B la ck e t al., J. B iol. C hem ., 267: 25513 -25520 , 1992. E stas a c il-C o A s in ta sas eu ca rio tas , a p e sa r de ca re c e r de a lta ho m o lo g ia con s e cu e n c ia s FadD de E. coli, pu ed en co m p le m e n ta r la ac tiv idad de FadD en F adD in ac tiva dos de E. coli.

How do I know how to do it? What are you doing here? Do you want to use it to use it to get to go here? OA, as well as riva two of common acids, include sc rita s in S ho cke yet al., P la nt P hys io l., 129: 1710-1722, 2002 ( Arabidopsis) , C av ig lia et al., J. B iol. C hem., 279: 1163-1169, 2004 (ra ta) and in K no ll et al., J. Biol. C hem., 269 (23): 16348-56, 1994 (le vad u ra). In the technique, it is known that the genes that encode these rates. See, po re jem p lo, Jo hn so net al., J. B io l. C hem., 269: 18037-18046, 1994; S ho cke yet al., P lan t P hys io l., 129: 1710-1722, 2002; B la ck et al., J. B iol. C hem., 267: 25513-25520, 1992. E stas ac il-C o A s in ta sas eu ca rio tas, ap r e r r of ca l cer de lta ho mo lo gu ia con se cu en ia s FadD of E. coli, may co-ment ment the activity of FadD in F adD in ac tiva two of E. coli.

A. Acyl-CoA en alcohol graso

A. Acyl-CoA in fatty alcohol

1. T ra n s fo rm a c io n de a c il-C o A en a lcoh o l g raso

1. T ra n s fo rm a c io n de a c il-C or A en a lcoh o l g sato

La a c il-C o A se reduce a un a lde h ido g ra so m ed ia n te una a c il-C o A red u c ta sa de pe n d ie n te de N A D H (p o r e jem p lo , A cr1 ). D espues, el a lde h ido g raso se reduce a un a lco h o l g ra so m ed ia n te una a lco h o l d e sh id ro g e n a sa d e pe nd ien te de N A D P H (p o r e jem p lo , Y qhD ). C om o a lte rna tiva , la a c il-C o A re d u c ta sa fo rm a d o ra de a lcoh o l g ra so (F A R ) ca ta liza la reducc ion de una a c il-C o A en un a lcoh o l g raso y C oA S H . La FAR utiliza N A D H o N A D P H com o un c o fa c to r en esta red ucc ion de cua tro e lec tron es . A u n q u e las rea cc io nes de FA R que ge ne ran a lcoh o l p roceden a tra ve s de un p ro d u c to r in te rm ed io de a ldeh ido , no se libe ra un a ld e h id o libre. P or tan to , las FAR fo rm a d o ra s de a lcoh o l son d is tin ta s de las e n z im a s que llevan a cabo red u cc io n e s de dos e lec tron es de a c il-C o A y que p roducen a lde h ido g ra so libre com o producto . (V ea se C heng y R usse ll, J. Biol. C hem ., 279 (36 ):37789 -37797 , 2004 ; M etz e t al., P lan t P hys io l., 122 :635-644 , 2000).

The ac-C or A is reduced to one of the greatest interest in the network of the NADH network (for example, A cr1 ). Then, the age of the private sector is reduced to one year or more than one year of the age of N A D P H (p o r e jem p lo, Y qhD). C om o lte rna tiva, the ac il-C or A re duc ta sa fo rm ado ra of a lcoh olg ra so (FAR) can reduce the reduction of an ac il-C or A in a lcoh olg satin and C oA SH. The FAR uses N A D H or N A D P H com o a c o fa c to r in this network network of four languages. Even if the FA R reactions that generate lcoh olp rubbed through a du p du du r in te rm ed io de a ldeh ido, a free year is not released . For example, the FAR FORMS of a lcoh ol are two of the enzymes that carry out networks of two e lec tron are of ac il-C or A and which produce the product as free as the product. (See C heng and R usse ll, J. Biol. C hem., 279 (36): 37789-37797, 2004; M etz et al., P lan t P hys io l., 122: 635-644 , 2000).

2. M od ifica c ion de las ru tas b io s in te tica s de ac ido s g ra sos para a u m e n ta r la tra n s fo rm a c io n de a c il-C o A en a lcoho l g ra so

2. M od ifica tion of the basic routes of interest in order to improve the work of the country or A in a few days.

Los h o sp e d a d o re s de p ro du cc ion pueden g e n o m a n ip u la rse u tiliza n d o po lip ep tido s c o n o c id o s para p ro d u c ir a lcoh o le s g ra so s a p a rtir de ac il-C o A . Un m e to d o pa ra p ro d u c ir a lco h o le s g ra so s im p lica a u m e n ta r la e xp re s io n de, o e xp re sa r fo rm a s m as ac tiva s de, a c il-C o A re d u c ta sa s fo rm a d o ra s de a lcoh o l g ra so (c o d ific a d a s po r un gen ta l co m o acr1, EC 1.2.1.50 /1.1.1 ), a c il-C o A re d u c ta sa s (E C 1.2.1.50 ) y /o a lcoh o l d e sh id ro g e n a sa s (E C 1.1.1.1).

The hoped e o r ents of p ro du cc ion can be used ip u la rse u use ndo po lip ep tido sconoc id os for p ro duc go to lcoh o le sg ra so sa sa rtir de ac il-C o A. A method for going to go to lco ho le sg ra so s im p lica increases ta r la e xp re s io n de, oe xp re sa r fo rm asm more ac tiva s de, ac il -C o A du du ta sa s fo rm ado ra s of a lcoh olg ra so (cod ific adas po r a gene ta l co mo acr1, EC 1.2.1.50 /1.1.1), ac il-C or A re duc ta sa s (EC 1.2.1.50) and / or lcoh olde sh id ro gena sa s (EC 1.1.1.1).

Los alcoholes grasos se describen a menudo como tensioactivos basados en hidrocarburos. Tambien sirven como componentes adecuados de tensioactivos. Para la produccion de tensioactivos, el hospedador de produccion se modifica para que produzca un tensioactivo a partir de una fuente de carbono renovable. Dicho hospedador de produccion incluye una primera secuencia de polinucleotidos exogena que codifica una proteina capaz de transformar un acido graso en un aldehido graso y una segunda secuencia de polinucleotidos exogena que codifica una proteina capaz de transformar un aldehido graso en un alcohol. En algunos ejemplos, la primera secuencia de polinucleotidos exogena codifica una reductasa de acido graso. En un ejemplo, la segunda secuencia de polinucleotidos exogena codifica la aldehido reductasa microsomal de mamifero o la aldehido deshidrogenasa de cadena larga. En un ejemplo adicional, la primera y la segunda secuencia de polinucleotidos exogena son de Arthrobacter AK 19, Rhodotorula glutinins, Acinetobacter sp. cepa M-1 o Candida lipolytica. En un aspecto, la primera y la segunda secuencia de polinucleotidos heterologa forman un complejo multienzimatico de Acinetobacter sp. cepa M-1 o de Candida lipolytica.

Fatty alcohols are often described as hydrocarbon-based surfactants. They also serve as suitable components of surfactants. For the production of surfactants, the production host is modified to produce a surfactant from a renewable carbon source. Said production host includes a first exogenous polynucleotide sequence encoding a protein capable of transforming a fatty acid into a fatty aldehyde and a second exogenous polynucleotide sequence encoding a protein capable of transforming a fatty aldehyde into an alcohol. In some examples, the first exogenous polynucleotide sequence encodes a fatty acid reductase. In one example, the second exogenous polynucleotide sequence encodes the mammalian microsomal aldehyde reductase or the long chain aldehyde dehydrogenase. In a further example, the first and second exogenous polynucleotide sequence are from Arthrobacter AK 19, Rhodotorula glutinins, Acinetobacter sp. strain M-1 or Candida lipolytica. In one aspect, the first and second heterologous polynucleotide sequences form a multienzyme complex of Acinetobacter sp. strain M-1 or Candida lipolytica.

Como fuentes adicionales de secuencias de ADN heterologas que codifican proteinas transformadoras de acidos grasos a alcoholes de cadena larga que pueden utilizarse en la produccion de tensioactivos se incluyen, pero sin limitacion, Mortierella alpina (ATCC 32222), Cryptococcus curvatus, (denominado tambien Apiotricum curvatum), Alcanivorax jadensis (T9T = DSM 12718 = At c C 700854), Acinetobacter sp. HO1-N (ATCC 14987) y Rhodococcus opacus (PD630 DSMZ 44193).

Additional sources of heterologous DNA sequences encoding fatty acid transforming proteins to long chain alcohols that can be used in the production of surfactants include, but are not limited to, Alpine Mortierella (ATCC 32222), Cryptococcus curvatus, (also called Apiotricum curvatum) ), Alcanivorax jadensis (T9T = DSM 12718 = At c C 700854), Acinetobacter sp. HO1-N (ATCC 14987) and Rhodococcus opacus (PD630 DSMZ 44193).

En un ejemplo, el derivado de acido graso es un producto tensioactivo saturado o insaturado que, en la cadena de carbono, tiene una longitud de aproximadamente 6 a aproximadamente 36 atomos de carbono, de aproximadamente 8 a aproximadamente 30 atomos de carbono, de aproximadamente 10 a aproximadamente 26 atomos de carbono, de aproximadamente 12 a aproximadamente 20 atomos de carbono, o de aproximadamente 12 a aproximadamente 16 atomos de carbono. En otro ejemplo, el producto tensioactivo tiene una longitud de aproximadamente 10 a aproximadamente 18 atomos de carbono, o de aproximadamente 12 a aproximadamente 14 atomos de carbono en la cadena de carbono.

In one example, the fatty acid derivative is a saturated or unsaturated surfactant product that, in the carbon chain, has a length of about 6 to about 36 carbon atoms, about 8 to about 30 carbon atoms, about 10 at about 26 carbon atoms, from about 12 to about 20 carbon atoms, or from about 12 to about 16 carbon atoms. In another example, the surfactant product has a length of about 10 to about 18 carbon atoms, or about 12 to about 14 carbon atoms in the carbon chain.

Los hospedadores de produccion adecuados para producir tensioactivos incluyen microorganismos eucariotas o procariotas. Como hospedadores de producciones ejemplares se incluyen Arthrobacter AK 19, Rhodotorula glutinins, Acinetobacter sp. cepa M-1, Arabidopsis thaliana, Candida lipolytica, Saccharomyces cerevisiae, cianobacterias, tales como, Synechocystis spp. y Synechococcus spp., algas, tales como, Clamydomonas, y bacterias de E. coli genomanipuladas para sobreexpresar acetil-CoA carboxilasa. Tambien pueden utilizarse hospedadores de produccion que demuestran una capacidad innata para sintetizar altos niveles de precursores de tensioactivos en forma de lipidos y aceites, tales como Rhodococcus opacus, Arthrobacter AK 19, Rhodotorula glutinins, E. coli, genomanipuladas para expresar acetil CoA carboxilasa, y otras cianobacterias oleaginosas, bacterias, levaduras y hongos.

Suitable production hosts for producing surfactants include eukaryotic or prokaryotic microorganisms. Hosts of exemplary productions include Arthrobacter AK 19, Rhodotorula glutinins, Acinetobacter sp. strain M-1, Arabidopsis thaliana, Candida lipolytica, Saccharomyces cerevisiae, cyanobacteria, such as, Synechocystis spp. and Synechococcus spp., algae, such as, Clamydomonas, and E. coli bacteria genomanipulated to overexpress acetyl-CoA carboxylase. Production hosts can also be used that demonstrate an innate ability to synthesize high levels of surfactant precursors in the form of lipids and oils, such as Rhodococcus opacus, Arthrobacter AK 19, Rhodotorula glutinins, E. coli, genomanipulated to express acetyl CoA carboxylase, and other oleaginous cyanobacteria, bacteria, yeasts and fungi.

B. Alcoholes grasos a esteres grasos

B. Fatty alcohols to fatty esters

Los hospedadores de produccion pueden genomanipularse utilizando polipeptidos conocidos para producir esteres grasos de diversas longitudes. Un metodo para producir esteres grasos incluye aumentar la expresion de, o expresar formas mas activas de, uno o mas peptidos de alcohol O-acetiltransferasa (Ec 2.3.1.84). Estos peptidos catalizan la acetilacion de un alcohol al transformar una acetil-CoA y un alcohol en una CoA y un ester. En algunos ejemplos, los peptidos de alcohol O-acetiltransferasa pueden expresarse junto con peptidos de tioesterasa seleccionados, peptidos FAS, y peptidos formadores de alcohol graso, permitiendo asi controlar las longitudes de la cadena de carbono, los niveles de saturacion y los grados de ramificacion. En algunos casos, el operon bkd puede coexpresarse para producir precursores de acidos grasos ramificados.

Production hosts can be genomanipulated using known polypeptides to produce fatty esters of various lengths. One method of producing fatty esters includes increasing the expression of, or expressing more active forms of, one or more O-acetyltransferase alcohol peptides (Ec 2.3.1.84). These peptides catalyze the acetylation of an alcohol by transforming an acetyl-CoA and an alcohol into a CoA and an ester. In some examples, the alcohol O-acetyltransferase peptides can be expressed together with selected thioesterase peptides, FAS peptides, and fatty alcohol-forming peptides, thus allowing to control carbon chain lengths, saturation levels and branching degrees . In some cases, the bkd operon can be coexpressed to produce branched fatty acid precursors.

Tal como se usa en el presente documento, los peptidos de alcohol O-acetiltransferasa incluyen peptidos en la clasificacion de enzimas numero EC 2.3.1.84, asi como cualquier otro peptido capaz de catalizar la transformacion de una acetil-CoA y un alcohol para formar una CoA y un ester. Adicionalmente, un experto en la materia apreciara que los peptidos de alcohol O-acetiltransferasa tambien pueden catalizar otras reacciones.

As used herein, the O-acetyltransferase alcohol peptides include peptides in the classification of enzymes number EC 2.3.1.84, as well as any other peptide capable of catalyzing the transformation of an acetyl-CoA and an alcohol to form a CoA and an ester. Additionally, one skilled in the art will appreciate that O-acetyltransferase alcohol peptides can also catalyze other reactions.

Por ejemplo, algunos peptidos de alcohol O-acetiltransferasa pueden aceptar otros sustratos ademas de los alcoholes grasos y/o tioesteres de acetil-CoA, tales como otros alcoholes y otros tioesteres de acil-CoA. Por lo tanto, tambien se incluyen dichos peptidos de alcohol O-acetiltransferasa no especificos o de divergencias especificas. Diversas secuencias peptidicas de alcohol O-acetiltransferasa estan disponibles al publico. En la tecnica se conocen ensayos para medir la actividad de los peptidos de alcohol O-acetiltransferasa. Ademas, las O-aciltransferasas pueden genomanipularse para impartir nuevas actividades y/o especificidades para el grupo acilo donador o el grupo alcohol aceptor. Las enzimas genomanipuladas pueden generarse a traves de estrategias logicas y evolutivas bien documentadas.

For example, some O-acetyltransferase alcohol peptides may accept other substrates in addition to fatty alcohols and / or acetyl-CoA thioesters, such as other alcohols and other acyl-CoA thioesters. Therefore, said non-specific O-acetyltransferase or specific divergent alcohol peptides are also included. Various peptide sequences of alcohol O-acetyltransferase are available to the public. Assays for measuring the activity of the O-acetyltransferase alcohol peptides are known in the art. In addition, the O-acyltransferases can be genomanipulated to impart new activities and / or specificities for the donor acyl group or the acceptor alcohol group. Genomanipulated enzymes can be generated through well-documented logical and evolutionary strategies.

C. Acil-CoA a Esteres Grasos

C. Acil-CoA to Fatty Esters

1. Produccion de esteres grasos

1. Production of fatty esters

Los es te res g ra sos se s in te tizan m ed ia n te una ac il-C oA : a lcoh o l a c iltra n s fe ra sa g rasa (p o r e jem p lo , e s te r s in tasa), que co n ju g a un a lco h o l g ra so de ca d e n a la rga con una a c il-C o A g ra sa m e d ia n te un e n la ce este r. Las e s te r s in ta sa s y los genes co d ifica n te s son co n o c id o s a p a rtir de la p lan ta de jo jo b a y la bacte ria . Acinetobacter sp. cepa ADP1 (a n te rio rm e n te Acinetobacter calcoaceticus ADP1). La e s te r s in ta sa ba c te ria na es una e n z im a b ifu nc iona l, que m ue s tra ac tiv ida d e s te r s in ta sa y la cap ac idad de fo rm a r tr ia c ilg lice ro le s a p a rtir de sus tra to s de d ia c ilg lice ro l y ac il-C o A g ra sas (ac tiv ida d ac il-C oA : d ig lice ro l a c iltra n s fe ra sa (D G A T )). El gen wax/dgat cod ifica ta n to la e s te r s in tasa co m o la D G A T. V ea se C heng et al., J. Biol. C hem ., 279 (36 ):37798 -37807 , 2004; K a lsch e u e r y S te in bu che l, J. Biol. C hem ., 278 :8075 -8082 , 2003. Las e s te r s in ta sa s ta m b ie n pu eden u tiliza rse pa ra p ro d u c ir c ie rto s es te re s g ra so s que pu ed en u tiliza rse com o com b us tib le , ta l com o b iod iese l, com o se d e sc rib e en este docum e n to .

These are the most important things to do here: a lcoh olac iltra ns fe ra sa g rasa (pore jem p lo, e s te rs in rate), which co n play a lazy game of cana de la rga with an ac il-C o A g ra sa in the middle of this one r r. These are the coding genes that are known as the joke of the jo jo bay bay. Acinetobacter sp. strain ADP1 (before rm in Acinetobacter calcoaceticus ADP1). This is an international institution, which is an enzyme that has been carried out and the capital of the Spanish Federation c ilg lice ro will take you out of your day's work c ilg lice ro ly ac il-C o A g ra sas (ac tiv ida d ac il-C oA: d ig lice ro lac iltra ns fe ra sa (DGAT)). The wax / dgat gene encodes this rate at the rate such as DGA T. V e se C heng et al., J. Biol. C hem., 279 (36): 37798-37807, 2004 ; K a lsch euery S te in bu che l, J. Biol. C hem., 278: 8075-8082, 2003. Those who are able to use it can be used to leave it for you. Let me know how you can tell me how to use it as well as how to explore it in this document.

2. M od ifica c ion de la ruta b ios in te tica de ac ido s g ra sos para p ro d u c ir e s te re s g ra sos u tiliza n d o e s te r s in ta sa

2. M od ifica tion of the route b ios in te tica de ac a s g ra sos para p ro d u c go e s re s g ra sos u tiliza n d o e s te r s in ta sa

La p ro du cc ion de es te res g rasos, in c luye nd o ceras, a p a rtir de a c il-C o A y a lcoh o le s , puede g e n o m a n ip u la rse u tilizan do po lip e p tid o s con oc idos . Un m etod o para p ro d u c ir e s te re s g ra sos in c luye a u m e n ta r la exp res ion de, o e x p re s a r fo rm as m as ac tiva s de, una o m as e s te r s in ta sas (E C 2.3.1.20 , 2.3.1.75 ). D ive rsas se cu e n c ia s pep tid icas de e s te r s in ta sa estan d isp o n ib le s al pub lico . En la pa ten te de E s tad os U n idos N° 7.118.896 , se p ro po rc ion an m e tod os para d e te rm in a r la ac tiv ida d de la e s te r s in tasa .

The protection of these resources, whether or not waxes, can be separated from ac c-C or A and lcoh or le s, can be used or used or used as a lipid. with ears. A method for you to go to you will continue to be in order to increase the expiration of the most active activities of one or more of you. sas (EC 2.3.1.20, 2.3.1.75). D ive rsas is aware of the public relations of the public. In the country of United States No. 7,118,896, it is possible to pay for the current activity of the rate in question.

En de te rm in a d o s aspec tos , si el p ro du c to de sea do es un b io co m b u s tib le basado en este r, puede m o d ifica rse un h o s p e d a d o r de p ro du cc ion de ta l m a n e ra que p ro d u zca un e s te r a p a rtir de una fue n te de ene rg ia renovab le . D icho h o s p e d a d o r de p ro du cc ion in c luye genes e xo ge no s que cod ifica n una e s te r s in ta sa que se exp resa para co n fe r ir a d icho h o s p e d a d o r de p ro d u cc io n la ca p a c id a d de s in te tiz a r un e s te r graso , sa tu ra do , in sa tu ra d o o ra m ifica d o a p a rtir de una fuen te de e n e rg ia renovab le . En a lgu nos aspec tos , el o rg a n ism o ta m b ie n puede e x p re s a r genes que co d ifica n las s ig u ie n tes p ro te inas e jem p la re s : e lo n g a sa s de ac ido s g rasos, a c il-C o A red uc tasa s , ac iltran s fe rasas , e s te r s in tasas, ac il tra n s fe ra sa s grasas, d iac ilg lice ro l a c iltran s fe rasas , t io e s te ra sa s y/o a c il-co A a lcoho l a c iltra n s fe ra s a s de cera . En un a sp e c to a lte rn a tivo , el o rg a n is m o e xp re sa un gen que co d ifica una e s te r s in ta sa /a c il-C o A :d ia c ilg lice ro l a c iltra n s fe ra sa b ifunc iona l. P o r e jem p lo , la e s te r s in ta sa /a c il-C o A :d ia c ilg lice ro l a c iltra n s fe ra sa b ifu nc iona l pu ede se le cc io n a rse de los co m p le jo s m u ltie n z im a tico s de Simmondsia chinensis, Acinetobacter sp. cepa ADP1 (a n te rio rm e n te Acinetobacter calcoaceticus ADP1), Alcanivorax borkumensis, Pseudomonas aeruginosa, Fundibacter jadensis, Arabidopsis thaliana o Alcaligenes eutrophus (p o s te rio rm e n te ren o m b ra d a com o Ralstonia eutropha). En un aspecto , para p ro d u c ir es te res, ta le s com o e s te re s de cera o grasos, las ac ido g raso e longasas, a c il-C o A red uc tasa s o cera s in tasas, se ob tienen y /o p roceden de un com p le jo m u ltie n z im a tico de Alcaligenes eutrophus (p o s te rio rm e n te re n o m b ra d a com o Ralstonia eutropha) o de o tros o rg a n ism o s c o n o c id o s en la b ib liog ra fia .

In terms of specific aspects, if the property is a bus based on it, a host can be modified so that ro du zca one of you rapa rtir of a source of ene rg ia renovab le. The host host of the pipeline in c luye genes e xo ge no s that encodes one of these teas to be sent for the purpose of going to host the host host The peace of mind in one of the countries is fatty, in your mind, in the best sense of being part of a renewal energy source. In some respects, the r an an ism or ta mb ie n can express genes that encode the s te s u s u s s te p s te sins e jem p la re s: e lo nga sa s of ac i sg rasos, ac il-C or A red uc rate s, ac iltran s ras rasas, es s rs in rates, ac iltra ns faith ra sa s fats, d iac ilg lice ro lac iltran s fe rasas This is te s sa sa / oac il-co A a lcoho lac iltra ns fe ra sas de wax. In a sp e ct a lte rn a tivo, the o rg an is moe xp re sa a gene that encodes a te srs in ta sa / ac il-C or A: d ia c ilg lice ro lac iltra ns fe ra sa b ifunc iona l. P o jem p lo, this is the case in which the ac-c or A: day of the licentiate of the illustrious faith in which he can be born from the co mp le jo smu ltie nz imAtico s of Simmondsia chinensis, Acinetobacter sp. strain ADP1 (before rm in te Acinetobacter calcoaceticus ADP1), Alcanivorax borkumensis, Pseudomonas aeruginosa, Fundibacter jadensis, Arabidopsis thaliana or Alcaligenes eutrophus (after rm in te r omb ra da com o Ralstonia eutropha). In one aspect, for this purpose, such as wax or fatty, greasy acids and longasas, ac il-C or A red uc rate so waxes in rates , they are obtained and / or touched by an immatical comrade of Alcaligenes eutrophus (after the rm in the rename of Ralstonia eutropha) or of others or rg an ism osconoc id os in the bio liog ra trust.

C o m o fu e n te s ad ic io n a le s de s e cu e n c ia s de A D N he te ro lo g a s que cod ifica n p ro te inas de s in tes is de e s te r u tiles en la p ro du cc ion de es te res g ra sos se inc luyen , pero sin lim itac ion , Mortierella alpina (p o r e jem p lo , A T C C 32222), Cryptococcus curvatus (tam b ien d e n o m in a d a Apiotricum curvatum), Alcanivorax jadensis (p o r e jem p lo , T 9T =D S M 12718 = A T C C 700854 ), Acinetobacter sp. HO1-N, (p o r e jem p lo , A T C C 14987) y Rhodococcus opacus (p o r e jem p lo , P D 630 , D S M Z 44193).

How do you know about DNA DNA? What is the gas that codes us? Do you know how to use it? These are included, but without limitation, Alpine Mortierella (for example, ATCC 32222), Cryptococcus curvatus (also called Apiotricum curvatum), Alcanivorax jadensis (for jem p lo , T 9T = DSM 12718 = ATCC 700854), Acinetobacter sp. HO1-N, (by jem p lo, ATCC 14987) and Rhodococcus opacus (pore jem p lo, PD 630, DSMZ 44193).

Los h o sp e d a d o re s de p ro du cc ion u tiles para p ro d u c ir es te res g ra sos pueden se r m ic ro o rg a n ism o s e u ca rio tas o p ro ca rio tas . C o m o e je m p lo s no lim ita n te s de h o sp e d a d o re s de p ro du cc ion para p ro d u c ir es te res g ra sos se inc luyen Saccharomyces cerevisiae, Synechococcus, Synechocystis, Clamydomonas, Candida lipolytica, E. coli, Arthrobacter AK19, Rhodotorula glutinins, Acinetobacter sp. cepa M-1, Candida lipolytica, y o tros m ic ro o rg a n ism o s o lea g in oso s .

The past few years of use for these services can be guaranteed or is also considered to be the most important options. How do you keep it? Do you want to know what is included in Saccharomyces cerevisiae, Synechococcus, Synechocystis, Clamydomonas, Candida lipolytica, E. coli, Arthrobacter AK19, Rhodotorula glutinins, Acinetobacter sp. strain M-1, Candida lipolytica, yo tros m ic ro or rg an ism oso lea g in oso s.

En un e jem p lo , se u tiliza la e s te r s in ta sa de Acinetobacter sp. ADP1 en el lo cus A A O 17391 (de sc rita en K a lsch e u e r y S te in bu che l, J. B io l. C hem ., 278: 8075 -8082 , 2003 ). En o tro e jem p lo , se u tiliza la e s te r s in ta sa de Simmondsia chinensis en el lo cus A A D 38041.

In an example, the use of Acinetobacter sp. ADP1 in lo cus AAO 17391 (from sc rita in K to lsch euery S te in bu che l, J. B io l. C hem., 278: 8075-8082, 2003). In other cases, the use of Simmondsia chinensis is used in ADA 38041.

En d e te rm in a d o s aspec tos , los es te res p ro d u c id o s segun los m e to d o s y las com p o s ic io n e s de este do cu m e n to , se sec re ta n o libe ran de las ce lu la s h o sp e d a d o ra s y, po r lo tan to , pueden recu p e ra rse ex tra ce lu la rm e n te . O p c io n a lm e n te , pu e d e u tiliza rse un e x p o rta d o r de es te res , ta l co m o un m ie m bro de la fa m ilia de FA TP , pa ra fa c ilita r la libe ra c io n de e s te re s en el e n to rn o ex tra ce lu la r. Un e je m p lo no lim ita n te de un e x p o rta d o r de e s te r a d e cu a d o es la p ro te ina tra n s p o rta d o ra de ac ido s g ra sos (ca d e n a la rga ) C G 7400 -P A , iso fo rm a A , de Drosophila melanogaster, en el lo cus N P _524723.

In terms of specific aspects, these are according to the methods and the com- positions of this document, it is said that they are not released from the ce However, they can, in return, be able to recover extra money. O p io na lm en te, it is possible to become an exhibitor of this, as a member of the FA TP family, to facilitate the liberation of freedom. One of these is in the first or third place. An employer does not limit him or her from an ex-president of his life when he is the person who is the leader of the city of Sg ra sos (cana de la rga) CG 7400 -PA, iso fo rm a A, by Drosophila melanogaster, in lo cus NP _524723.

D. Acil-ACP, Acil-CoA a Hidrocarburo

D. Acil-ACP, Acil-CoA to Hydrocarbon

1. H id ro ca rb u ro s de m ic ro o rg a n ism o s pa rticu la res

1. H id ro ca rb u ro s de m icro o rg a n ism o s pa rticu la res

S e con oce un con ju n to d ive rso de m ic ro o rg a n ism o s que p roducen h id ro ca rbu ros , ta le s com o a lcanos, o le fin as e iso p re no ide s . M uch os de estos h id ro ca rb u ro s p roceden de la b ios in tes is de ac ido s g rasos. La p ro du cc ion de estos h id ro ca rb u ro s puede co n tro la rse c o n tro la n d o los genes aso c ia d o s con la b ios in tes is de ac ido s g rasos en los h o sp e d a d o re s de p ro du cc ion na tivos.

I know that there is a difference in the number of people that is responsible for making them known, [...] as is the case, or the end of this idea. Many of these heroes fall from the bios ios tes de s a c gone s satin. The protection of these heroes can be linked to the genes associated with the two sources of succession in the homes of families. s of p ro du cc ion na tivos.

P o r e jem p lo , la b ios in tes is de h id ro ca rb u ro s en la espec ie de a lga Botryococcus braunii se p ro du ce m ed ia n te la d e sca rb o n ila c io n de a ld e h id o s grasos. Los a ld e h id o s g rasos se p ro du cen po r la re d u cc io n de ac il tio e s te re s g rasos m ed ia n te una e n z im a ta l com o la a c il-C o A red uc tasa grasa . P o r tan to , la e s truc tu ra de los a lcan os fin a le s puede c o n tro la rse g e n o m a n ip u la n d o la espec ie B. braunii pa ra e xp re sa r ge ne s esp ec ificos , ta le s com o los de las t io e s te ra sa s , que con tro la n la lo ng itud de la ca d e n a de los ac ido s g ra sos que se can a liza n en b ios in tes is de a lcanos. La exp res ion de las en z im a s que p ro du cen la b ios in tes is de ac ido s g ra sos de cad en a ram ificada en B. braunii da ra lu g a r a la p ro du cc ion de a lcan os de cad en a ram ificada . La in tro d u cc io n de genes que a fe c ta n a la p ro du cc ion de ac ido s g ra sos de sa tu ra d o s da ra com o resu ltad o la p ro du cc ion de o le finas . Las co m b in a c io n e s ad ic io n a le s de estos ge ne s pueden p ro p o rc io n a r un con tro l ad ic io n a l sob re la e s tru c tu ra fina l de los h id ro ca rb u ro s que se p roduc iran . P ara p ro d u c ir n ive les m as a lto s de h id ro ca rb u ro s na tivos o g e n o m a n ip u la d o s , los ge ne s im p lica d o s en la b ios in tes is de ac ido s g ra sos y sus p re cu rso re s , o en la d e g ra d a c io n de o tros p ro du c to s , pu ed en exp re sa rse , s o b re e x p re s a rs e o a te nua rse . C ada una de es ta s e s tra te g ia s pu ede a p lica rse a la p ro du cc ion de a lcan os en Vibrio fumissii cep a M1 y en o tras cepas de Vibrio fumissii, que p roducen a lcan os a tra ve s de la red ucc ion de a lcoh o le s grasos. A d e m a s de Vibrio fumissii, pueden u tiliza rse o tros o rg a n ism o s p ro d u c to re s de a lca n o s que u tilizan la ru ta de los ac ido s grasos. C a da uno de estas e s tra te g ia s ta m b ie n puede ap lica rse a la p ro du cc ion de o le finas p ro du c ida s po r cepas de Micrococo leuteus, Stenotrophomonas maltofila, y m ic ro o rg a n ism o s re lac io na do s . E stos m ic ro o rg a n ism o s p roducen o le fin a s de ca d e n a la rga que p ro ced en de la co n d e n sa c io n de cab eza a ca b e za de p re cu rso re s de ac ido s g ra sos . El con tro l de la e s truc tu ra y el n ive l de los p re cu rso re s de ac ido s g ra sos u tilizan do los m e tod os d e sc rito s en este d o cu m e n to da ran com o resu lta d o la fo rm a c io n de o le fin as de d ife re n te s lo ng itude s de cadena , ca ra c te ris tica s de ra m ifica c io n y n ive le s de sa tu rac ion .

P o jem p lo, b ios in tes is de h id ro ca rb u ro s in the spec a a lga Botryococcus braunii p p du du ce m ed ia n te la from sca rb on ila c io n a ld eh id os fatty. Those who have been open for the purpose of the re du ction of ac i tio e s te re sg ras m ed ia n t an enzyme im a ta l com o the ac il-C o A uc network fat rate. For the purpose of the years, the end of the year can be achieved by the genoman ip u nde the speci b B. braunii para e xp re sa r ge ne s esp ec ificos, ta The s of the uncles is te ra sa s, that with it the lo ng itud of the cane de los ac gone s s ra ra s that can be used in b ios tees de a lcanos . The expiration of the areas that have been the main interest of the United States in B. braunii will give rise to the protection of the acan os of cad in a ram ified. The introduction of genes that, according to the law, is only one of the results that will result in the resolution of the fine. The commissions in addition to these gens can be able to provide a final commitment to the final structure of the IDPs. u ro s that p roduc will go. P o p ro duc go n live les lto s de l id s of h id ro ca rb u ro s na tivos ogenoman ip u la dos, the ge ne s imp p lica two in the b ios in tes is de ac ac s sg So you and your parents, or in the course of the other days, you may want to expire, so that you will be dismissed. Each one of these strategies can be applied to the protection of Vibran fumissii cep a M1 and in or after Vibrio fumissii strains , which produce can through the network of a lcoh o le s fatty. In addition to Vibrio fumissii, they can be used by others or by the way they use the route of fatty acids. One of these strategies can also be applied to the protection of the strains of Micrococo leuteus, Stenotrophomonas maltofila, and my throats. or rg an ism os re lac io na do s. These m o rro or rg an ism osp or the end of the channel that allows the accession of the community of finance to the capital of the region. sg ra sos. The control of the structure and the level of the priests in which they are used are the methods of the rites in this period of time. or as a result of the aim of the end of the chain, this is the rationale for the rationale of the group. ion

Las c ia n o b a c te ria s ta m b ie n pueden u tiliza rse com o ho sp e d a d o re s de p ro du cc ion ad e cu a d o s para la p ro du cc ion de d e riva d o s de ac ido s g ra sos ta le s com o a lcoh o le s g rasos , es te res g ra sos e h id ro ca rbu ros . P o r e jem p lo , Synechocystis sp. P C C 6803 y Synechococcus elongatusPCC7942 pueden s e rv ir com o h o sp e d a d o re s de p ro du cc ion y pueden g e n o m a n ip u la rse u tilizan do te cn ica s de b io log ia m o le cu la r c o n ve n c io n a le s (Thiel, G e n e tic ana lys is o f cya n o b a c te ria , en T H E M O L E C U L A R B IO L O G Y O F C Y A N O B A C T E R IA , A D V A N C E S IN P H O T O S Y N T H E S IS A N D R E S P IR A T IO N 581-611 (K lu w e r A c a d e m ic P ub lishe rs ), 1994; K oksh a ro va y W o lk , A pp l. M icrob io l. B io techno l., 58: 123-137 , 2002. Los genes de b ios in tes is de ac ido s g ra sos pueden id en tifica rse y a is la rse fa c ilm e n te en estos o rg an ism os.

The nobac te ria s ta mb ie n can be used as a common source of income for the purposes of de riva dos de riva dos de acido gone sg ra sos ta If you like it or not, this is what you are saying. P ore jem p lo, Synechocystis sp. PCC 6803 and Synechococcus elongatusPCC7942 can be sent as a source of protection and can be used as a logotype for the logistic process. s ( Thiel, G ene tic ana lys is of cya nobac te ria, in THEMOLECULARB IO LOGYOFCYANOBACTER IA, ADVANCES IN PHOTOSYNTHES IS ANDRESP IR AT IO N 581-611 (K lu wer A cadem ic P ub lishe rs), 1994; K oksh a ro va and W o lk, A pp l. M icrob io l. B io techno l., 58: 123-137, 2002. The genes of b ios in tes is de ac ido sg ra sos can be in tifica Leaving is already the easy way in these or other countries.

A s im ism o , m uchas c ia n o b a c te ria s son p ro du c to ras na tu ra les de h id ro ca rbu ros , ta le s com o he p tad eca no , y po r lo ta n to con tien en genes de b ios in tes is de h id ro ca rb u ro s que pueden d e s re g u la rse y so b re e xp re sa rse ju n to con la m an ip u la c ion de sus ge ne s de b ios in tes is de ac ido s grasos, para a u m e n ta r la p ro du cc ion de h id ro ca rbu ros .

A s im ism o, many of them will be the only ones that have a bad reputation, not as well as not, and not so much. In this way, in the genes of the countries that can be explained and that they can be left with the same way as their Genetics of fatty acids, in order to increase the risk of human rights.

A d ife ren c ia de o tras bacte rias , a lgu nas c ia n o b a c te ria s (p o r e jem p lo , Synechocystis sp. P C C 6803 ) co n tie n e n ac idos g ra sos p o liin sa tu ra d o s en sus lip ido s (M ura ta , P la n t C e ll P hys io l., 33: 933 -941 , 1992), y po r lo ta n to tie n e n la cap ac idad in trinse ca de p ro d u c ir d e riva d o s de ac ido s g ra sos po liin sa tu rad os . Lo que es m as im portan te , las c ia n o b a c te ria s son o rg a n ism o s fo to s in te tico s que s in te tizan to d o el ca rb o n o c e lu la r reco g iend o luz s o la r y fija nd o d iox id o de carbono . P o r lo tan to , los de riva dos de ac ido s g ra sos p ro du c ido s en c ia n o b a c te ria s , p roceden d ire c ta m e n te de C O ².

A d ife ren c ia de o after bacte rias, to some of them nobac te r s (pore jem p lo, Synechocystis sp. PCC 6803) where they have acidic acids in their lives lipid s (M ura ta, P la nt C e ll P hys io l., 33: 933-941, 1992), and so they can have the capital in trinse ca de p ro duc go from riva dos de ac gone sg ra sos po liin sa tu rad os. What is most important, the nobac te ria s are or rg an ism os fo to s in teticos s that in te tized to the ca rb onoce lu the r reco g iend or light so the ry fixed nd od iox id o carbon. For the most part, those of Riva Dos de Ac Ido Sg Ra Sos Pro Du C gone s in the New York City, P roceden d ire C ta Men T of CO ² .

2. P rod ucc ion de h id ro ca rb u ro s a p a rtir de la red ucc ion de a lcoh o le s p rim arios

2. P r u u rion of h id ro ca rb u ro s a p artir of the ucc ion network of a lcoh o le s p rim arios

Los h id ro ca rbu ros ta m b ie n pueden p ro du c irse u tilizan do o x id o rre d u c ta sa s e vo lu c io n a d a s para la reducc ion de a lco h o le s p rim arios . S e con oce la u tilizac ion de a lcoh o le s g rasos p rim a rio s para p ro d u c ir a lcan os en m ic ro o rg a n ism o s , ta le s com o Vibrio fumissii M1. V ease , po r e jem p lo , Park, J. B acte rio l., 187 :1426 -1429 , 2005. Un e je m p lo de una o x id o rre d u c ta sa que puede u tiliza rse para p ro d u c ir h id ro ca rb u ro s a p a rtir de a lco h o le s g ra sos es la o x id o rre d u c ta sa d e pe nd ien te de N A D (P )H . Las o x id o rre d u c ta sa s s in te ticas de p e n d ie n te s de N A D (P )H pueden p ro du c irse u tilizan do g e n o m a n ip u la c io n evo lu tiva y pueden exp re sa rse en ho sp e d a d o re s de p roducc ion para p ro d u c ir d e riva d o s de ac ido s grasos.

Those who are able to use them can be used for the purpose of reducing them in order to reduce the amount of money they receive. I know the use of a lcoh or le sgs p rim a rio s for p du du go to lcan os in m icro or rg an ism os, as le s com o Vibrio fumissii M1. See, po re jem p lo, Park, J. B acte rio l., 187: 1426-1429, 2005. An example of an ox id or rre duc ta sa that can be used to go for du duc go h id ro ca rb u ro sapa rtir de a lco ho le sg ra sos is the ox id o rre duc ta sa de pe nd ien te de NAD (P) H. The oxidized du r ta du s ss in pend e n t s techniques of NAD (P) H can be used for genoman ip u and the positive evo ive and can be expressed in Ho sp edado re s de p roducc ion for p ro duc ir de riva two of fatty acids.

Los exp e rtos en la te cn ica conocen y lleva a la p ra c tica el p ro ceso de "co n ve rtir" una a lcoh o l red u c ta sa g rasa para te n e r la ac tiv idad de se a d a (K o lkm a n y S tem m er, Nat. B io techno l., 19 :423 -8, 2001 ; N ess et al., A dv. P ro te in C hem ., 55 :261 -92 , 2000; M in shu ll y S tem m er, C urr. O pin . C hem . B io l., 3 :284 -90 , 1999; H u ism an y G ray, C urr. Opin. B io te chn o l., 13 :352-8 , 2002; p u b licac ion de pa ten te de E s tad os U n idos n.° 2006 /0195947.

The experts in the technical field know and carry out the practice of "co n ve rtir" a la l'coh ol red uc ta sa g rasa in order to have the activity of the se ( K o lkm any S tem m er, Nat. B io techno l., 19: 423-8, 2001; N ess et al., A dv. P ro te in C hem., 55: 261-92, 2000; M in shu ll and S tem m er, C urr. Or pin. C hem. B io l., 3: 284-90, 1999; H u ism an and G ray, C urr. Opin. B io te chn o l., 13: 352-8, 2002; public license publication of the United States No. 2006/0195947.

U na b ib lio teca de o x id o rre d u c ta sa s de p e n d ie n te s de N A D (P )H se ge ne ra m ed ia n te m e to d o s es tanda r, ta le s com o P C R p ro p e n sa a error, m u ta g e n e s is a le a to ria e sp ec ifica de sitio , m u ta g e n e s is de sa tu ra c io n e sp e c ifica de sitio o m u ta g e n e s is e sp ec ifica d irig ida a s itio . A d ic io n a lm e n te , puede c rea rse una b ib lio teca m ed ia n te el "b a ra ja d o " de s e cu e n c ia s co d ifica n te s de la o x id o rre d u c ta sa d e p e n d ie n te de N A D (P )H de o rigen na tu ra l. La b ib lio te ca se exp resa en un h o sp e d a d o r de p ro du cc ion adecu ad o , ta l com o E. coli. Las co lo n ia s in d iv id u a le s que exp resan un m ie m bro d ife re n te de la b ib lio teca de o x id o rre d u c ta sa se an a lizan de spu es para d e te rm in a r la exp res ion de una o x id o rre d u c ta sa que pued e c a ta liz a r la red ucc ion de un a lcoh o l graso.

U na b ib lio teca de ox id o rre duc ta sa s de pend ie n te s de NAD (P) H is generated in two months is tanda r, ta le s com o PCR P ro pen sa a error, mu ta g e nes is a le a ria e sp site ec, mu ta g e nes is de sa tu ra c io ne sp e site ec omu ta g e nes is e sp ec irica d irig ida as itio. On a daily basis, you can use a bi- lio teca m ed ia n the "ba ra ja do" of the code of the ox id or rre duc ta It depends on NAD (P) H of or govern na tu ra l. The b ib lio te ca expires in an adequate age of adequate protection, such as E. coli. The co ia ns in d iv id ua le s that exp m a m ie m bro d ife re n te of the b ib lio teca de ox id o rre duc ta sa are analyzed from spu es para de te rm in ar the expiration of an ox id or rre duc ta sa that can be used for the fatty network.

P o r e jem p lo , cad a ce lu la puede e n sa ya rse com o una b io tra n s fo rm a c io n de ce lu la s com p le ta s , un e x tra c to ce lu la r, o una celula permeabilizada. Tambien pueden analizarse enzimas purificadas de la celula. Las alcohol reductasas grasas se identifican mediante un control espectrofotometrico o fluorometrico de la oxidacion dependiente de alcohol graso de NAD(P)H. La produccion de alcanos se controla mediante GC-MS, TLC u otros metodos adecuados.

P o jem p lo, you can find it in a common way, or you can find it, you can find it, an ex-tra c t ce ce r, or a permeabilized cell. Purified enzymes from the cell can also be analyzed. The fatty alcohol reductases are identified by a spectrophotometric or fluorometric control of the fatty alcohol dependent oxidation of NAD (P) H. The production of alkanes is controlled by GC-MS, TLC or other suitable methods.

Una oxidorreductasa identificada de esta manera se utiliza para producir alcanos, alquenos e hidrocarburos ramificados relacionados. Esto se logra ya sea in vitro o in vivo. Este ultimo se logra expresando el gen de la alcohol reductasa grasa evolucionado en un organismo que produce alcoholes grasos, como los que se describen en el presente documento. Los alcoholes grasos actuan como sustratos para la alcohol reductasa, que produce alcanos. Otras oxidorreductasas tambien pueden genomanipularse para catalizar esta reaccion, como las que utilizan hidrogeno molecular, glutation, FADh u otras coenzimas reductoras.

An oxidoreductase identified in this way is used to produce alkanes, alkenes and related branched hydrocarbons. This is achieved either in vitro or in vivo. The latter is achieved by expressing the fatty alcohol reductase gene evolved in an organism that produces fatty alcohols, such as those described herein. Fatty alcohols act as substrates for alcohol reductase, which produces alkanes. Other oxidoreductases can also genomanipulate to catalyze this reaction, such as those using molecular hydrogen, glutathione, FADh or other reducing coenzymes.

3. Conversion de acil-ACP a cetona y/u olefinas

3. Conversion of acyl-ACP to ketone and / or olefins

La proteina transportadora de acilos, acil-ACP, puede transformarse en una cetona y/o en una olefina interna mediante la accion de enzimas condensadoras de acilo, como se describe en la Publicacion PCT n.° 2008/147781 A2. Como se describe en la publicacion '781, los peptidos condensadores de acilo incluyen peptidos que son capaces de catalizar la condensacion de acil-ACP, acil-CoA, acil-AMP, acidos grasos, y sus mezclas, utilizando los metodos descritos en la misma. En algunos aspectos, estos peptidos condensadores de acilo tienen una alta, media o baja especificidad de sustrato. En determinados ejemplos, los peptidos condensadores de acilo son mas especificos de sustrato y solo aceptaran sustratos de una longitud de cadena especifica. Adicionalmente, un experto habitual en la materia apreciara que algunos peptidos condensadores de acilo tambien catalizaran otras reacciones. En la publicacion '781 se desvelan ejemplos de enzimas condensadoras de acilo. Ademas, la publicacion '781 describe proteinas de adenilacion, deshidratasas y deshidrogenasas que pueden utilizarse en la produccion de hidrocarburos tales como olefinas internas.

The acyl transporter protein, acyl-ACP, can be transformed into a ketone and / or an internal olefin by the action of acyl condensing enzymes, as described in PCT Publication No. 2008/147781 A2. As described in publication '781, acyl condenser peptides include peptides that are capable of catalyzing the condensation of acyl-ACP, acyl-CoA, acyl-AMP, fatty acids, and mixtures thereof, using the methods described therein . In some aspects, these acyl condenser peptides have a high, medium or low substrate specificity. In certain examples, acyl condenser peptides are more substrate specific and will only accept substrates of a specific chain length. Additionally, a person skilled in the art will appreciate that some acyl condenser peptides will also catalyze other reactions. Examples of acyl condensing enzymes are disclosed in publication '781. In addition, publication '781 describes adenylation proteins, dehydrates and dehydrogenases that can be used in the production of hydrocarbons such as internal olefins.

Los organismos recombinantes pueden genomanipularse utilizando polinucleotidos y proteinas, por ejemplo, los divulgados en la publicacion '781, para producir hidrocarburos y cetonas alifaticas que tienen caracteristicas estructurales definidas (por ejemplo, grado de ramificacion, niveles de saturacion, o longitudes de la cadena de carbono). Un metodo para producir hidrocarburos implica aumentar la expresion de, o expresar formas mas activas de, una o mas enzimas condensadoras de acilo (enzimas que condensan dos o mas de acil-CoA, acil-ACP, acil-AMP, acil-ester, acido graso, o mezclas de los mismos). Un experto en la materia apreciara que los productos producidos a partir de dichas reacciones de condensacion varian dependiendo de la cadena de acilo que se condensa. Los productos que pueden producirse incluyen, por ejemplo, hidrocarburos y productos intermedios de hidrocarburo, tales como cetonas alifaticas. Las cetonas alifaticas, los hidrocarburos y los productos intermedios de hidrocarburo pueden genomanipularse para tener caracteristicas especificas de la cadena de carbono expresando diversas enzimas o atenuando la expresion de diversas enzimas en el organismo recombinante. Segun la presente divulgacion, las tioesterasas mutantes de la divulgacion pueden utilizarse para manipular el intervalo de longitudes de la cadena de carbono de las especies de acilo. Por tanto, utilizando una tioesterasa mutante que tenga una especificidad o selectividad de sustrato particular, es posible influir en las reacciones posteriores para dar como resultado un perfil de producto de olefina o cetona predeterminado.

Recombinant organisms can be genomanipulated using polynucleotides and proteins, for example, those disclosed in publication '781, to produce hydrocarbons and aliphatic ketones that have defined structural characteristics (e.g., degree of branching, saturation levels, or chain lengths of carbon). One method of producing hydrocarbons involves increasing the expression of, or expressing more active forms of, one or more acyl condensing enzymes (enzymes that condense two or more of acyl-CoA, acyl-ACP, acyl-AMP, acyl ester, acid fatty, or mixtures thereof). One skilled in the art will appreciate that the products produced from said condensation reactions vary depending on the acyl chain that is condensed. Products that can be produced include, for example, hydrocarbons and hydrocarbon intermediates, such as aliphatic ketones. Aliphatic ketones, hydrocarbons and hydrocarbon intermediates can be genomanipulated to have specific characteristics of the carbon chain expressing various enzymes or attenuating the expression of various enzymes in the recombinant organism. According to the present disclosure, the mutant thiosterases of the disclosure can be used to manipulate the range of carbon chain lengths of acyl species. Therefore, using a mutant thioesterase having a particular substrate specificity or selectivity, it is possible to influence subsequent reactions to result in a predetermined olefin or ketone product profile.

4. Transformacion de acido graso a aldehido

4. Transformation of fatty acid to aldehyde

Los acidos grasos resultantes de la escision de tioesterasa pueden transformarse en un aldehido mediante la accion del gen de la acido carboxilico reductasa. Los aldehidos pueden ser productos utiles en si mismos o pueden servir como sustratos para otras reacciones de catalisis enzimatica, por ejemplo, en la produccion de alcoholes grasos mediante una reaccion enzimatica de alcohol deshidrogenasa, o en la produccion de alcanos mediante una reaccion enzimatica de descarboxilasas. Segun las composiciones y los metodos desvelados en el presente documento, los sustratos de acidos grasos de la acido carboxilico reductasa pueden manipularse para lograr un perfil de producto predeterminado en el producto de aldehido o alcohol graso.

The fatty acids resulting from thioesterase cleavage can be transformed into an aldehyde by the action of the carboxylic acid reductase gene. The aldehydes can be useful products in themselves or they can serve as substrates for other enzymatic catalysis reactions, for example, in the production of fatty alcohols by an enzymatic reaction of alcohol dehydrogenase, or in the production of alkanes by an enzymatic reaction of decarboxylases . According to the compositions and methods disclosed herein, the carboxylic acid reductase fatty acid substrates can be manipulated to achieve a predetermined product profile in the aldehyde or fatty alcohol product.

E. Liberacion de derivados de acidos grasos - con o sin proteinas transportadoras

E. Release of fatty acid derivatives - with or without carrier proteins

Como se describe en este documento, los derivados de acidos grasos producidos segun los metodos, las composiciones, los vectores y las celulas hospedadoras de este documento, pueden secretarse o liberarse espontaneamente para permitir la recuperacion extracelular de los productos derivados de acidos grasos. La velocidad de la secrecion espontanea puede ser o no lo suficientemente rapida, y el nivel de liberacion puede ser o no lo suficientemente completo. Por lo tanto, opcionalmente, las proteinas transportadoras pueden utilizarse para facilitar la exportacion de derivados de acidos grasos fuera del hospedador de produccion. Se sabe que las proteinas transportadoras y de salida excretan una gran variedad de compuestos, y que pueden modificarse naturalmente para que sean selectivas para tipos de derivados de acidos grasos particulares. Son ejemplos no limitantes de proteinas transportadoras adecuadas las proteinas transportadoras de casetes de union a ATP (ABC), las proteinas de salida y las proteinas transportadoras de acidos grasos (FATP, fatty acid transporter proteins). Como ejemplos no limitantes adicionales de proteinas transportadoras adecuadas se incluyen las proteinas transportadoras de ABC de organismos tales como Caenorhabditis elegans, Arabidopsis thaliana, Alcaligenes eutrophus y Rhodococcus erythropolis. Como ejemplos de proteinas transportadoras de ABC se incluyen CER5, AtMRP5, AmiS2 o AtPGP1.

As described herein, fatty acid derivatives produced according to the methods, compositions, vectors and host cells herein, can be secreted or released spontaneously to allow extracellular recovery of fatty acid derivative products. The speed of spontaneous secretion may or may not be fast enough, and the level of release may or may not be complete enough. Therefore, optionally, the carrier proteins can be used to facilitate the export of fatty acid derivatives outside the production host. It is known that carrier and output proteins excrete a wide variety of compounds, and that they can be naturally modified to be selective for particular types of fatty acid derivatives. Non-limiting examples of suitable transport proteins are ATP-binding cassette transporter proteins (ABC), exit proteins and fatty acid transporter proteins (FATP ). Additional non-limiting examples of suitable transport proteins include ABC transport proteins from organisms such as Caenorhabditis elegans, Arabidopsis thaliana, Alcaligenes eutrophus and Rhodococcus erythropolis. Examples of ABC transport proteins include CER5, AtMRP5, AmiS2 or AtPGP1.

En un a sp ec to p re fe rido , la p ro te ina tra n s p o rta d o ra de A B C es una C E R 5 (p o r e jem p lo , A Y 734542 )). Los ve c to re s que con tien en genes que exp resan p ro te inas tra n sp o rta d o ra s ad e cu a d a s pueden in se rta rse en ho sp e d a d o re s de p ro du cc ion de p ro te in a s para a u m e n ta r o im p u lsa r la libe ra c io n de d e riva d o s de ac ido s grasos.

In one year, the first step of A B C is a C E R 5 (p o r e jem p lo, A Y 734542)). They see that they have in genes that expire for tra t sp o rs do ra s ad e when they can be inserted into ho ho e reed p o du r s of p ro du cc ion de p ro Teas to increase the importance of fatty acids.

La p ro du cc ion de p ro du c to s d e riva d o s de ac ido s g ra sos seg un la p re sen te d ivu lg a c io n no req u ie re la m od ifica c io n de la p ro te ina tra n s p o rta d o ra o de sa lid a y es po s ib le se le c c io n a r ho sp e d a d o re s de p ro d u cc io n po r su capac idad e n d o ge na pa ra lib e ra r d e riva d o s de ac ido s g rasos. A s im ism o , s im p lem e n te c o n s tru ye n d o ce lu la s h o spe da do ra s seg un la p re se n te d ivu lg a c io n , po r e jem p lo , p ro du c to s d e riva d o s de ac ido s g ra so s que, de o tra m anera , se sab e que no se secre tan , pueden se c re ta rse o libe ra rse e sp o n ta n e a m e n te . La e fica c ia de la p ro du cc ion y libe rac ion del p ro d u c to en el ca ldo de fe rm e n ta c io n puede e xp re sa rse com o una re lac io n en tre el p ro du c to in tra c e lu la r y el p ro d u c to ex trace lu la r. En a lgu nos e jem p los , la re lac ion puede s e r de a p ro x im a d a m e n te 100:1, 50:1, 20:1, 10:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:10, 1:20, 1:30, 1:40 o 1:50.

The p ro du ction of p ro du c to sde riva two of akg sg ra you are according to the p re sen te d ivu lg ac i n no req u ie re the m od ification of the p ro It will help you through your life and that is why you will be told about your status as a result of your end-to-end capacity for liberty. two of a century ago. A s im ism o, s im p lem en te constru ye ndo ce lu sho sho da do ra s seg un la p re se n te d ivu lg ac io, po re jem p lo, p ro du c All of this is the case that, in some other way, it is known that it is not secreted, it can be reduced or released in the same way. The effectiveness of the p du du tion and liberation of the p r duct in the state of faith rm in which it can be taken as a re ection in relation to the p ro du c to in tra ce lu la r and p ro duc to ex trace lu la r. In some cases, the relationship may be approximately 100: 1, 50: 1, 20: 1, 10: 1, 5: 1, 4: 1, 3: 1, 2: 1, 1: 1, 1: 2, 1: 3, 1: 4, 1: 5, 1:10, 1:20, 1:30, 1:40 or 1:50.

II. Seleccion de las caracteristicas de la cadena de carbono de los derivados de acidos grasos

II. Selection of the characteristics of the carbon chain of fatty acid derivatives

D e riva d o s de ac ido s g ra sos con pu n tos de ram ificac io n , n ive les de sa tu ra c io n , lo n g itu d e s de la ca d e n a de c a rb o n o y ca ra c te ris tica s de e s te r p a rticu la re s pu ed en p ro d u c irse seg un se desee . C o m o h o sp e d a d o re s de p ro du cc ion pueden se le cc io n a rse m ic ro o rg a n ism o s que, de m an e ra na tu ra l, p ro du cen d e riva d o s pa rticu la re s y, en c ie rta s c ircu ns tan c ias , las en z im a s en d o g e n a s en su in te rio r pueden m a n ip u la rse para p ro d u c ir d e riva d o s de ac ido s g ra sos de ca ra c te ris tica s deseab les . C om o a lte rna tiva , los genes que exp resa n e n z im a s que p roduc iran de riva dos de ac ido s g ra sos pa rticu la re s pueden in se rta rse a d e cu a d a m e n te en m ic ro o rg a n ism o s h o sp e d a d o re s de p roducc ion .

Two of the following are with ramifications, the levels of the channel, the channels of the chain of channels, and the rationale. Of this, it is necessary for you to be able to leave as you wish. How do you want to know if you can see that you will be able to do it for two days? s, in this sc ircu ns tan c ias, those in zin as in d in rio r r r can keep ip for the du riva de riva dos de acido sg ra sos de ca You want to laugh at them. C om o lte rna tiva, the genes that exp res nenz im so that p roduc will go from riva two to arc sg ra s pa rrticu re s s can be se su r se r se su su temmen in m ic ro or rg an ism osho sp edado re s de p roducc ion.

En a lgu nos e jem p los , la exp res ion de genes FA S exogenos , que se o rig in an a p a rtir de d ife ren te s esp ec ies o v a r ia n te s g e n o m a n ip u la d a s , pued e rea liza rse en un h o sp e d a d o r de p ro du cc ion , da n d o com o resu lta d o la b ios in tes is de a c id o s g ra sos que son e s tru c tu ra lm e n te d ife re n te s (p o r e jem p lo , en long itudes , n ive le s de ram ificac io n , g ra do s de in sa tu ra c ion , e tc .) a los de l h o sp e d a d o r de p ro du cc ion nativo. E stos p ro du c to s ge n icos h e te ro logo s ta m b ie n pueden se le cc io n a rse o g e n o m a n ip u la rse para que no se vean a fe c ta dos po r los m e ca n ism o s reg u la d o re s na tu ra les en las ce lu la s h o sp e d a d o ra s de p ro du cc ion , y com o ta l p e rm itir el con tro l de la p ro d u cc io n de l p ro d u c to c o m e rc ia l deseado . P o r e jem p lo , las e n z im a s FA S de Bacillus subtilis, Saccharomyces cerevisiae, Streptomyces spp., Ralstonia, Rhodococcus, Corynebacteria, Brevibacteria, Mycobacteria, le vad u ras o lea g in osa s , o s im ila res , pueden exp resa rse en un h o s p e d a d o r de p ro du cc ion adecu ad o . La exp res ion de d ich a s en z im a s exo ge na s a lte ra ra la e s truc tu ra del a c id o g ra so p roduc ido .

In some cases, the expiration of exogenous FA S genes, which are known to be possible, will be possible for the next few days, may be carried out. Go in a ho o age of p ro du cc ion, as a result of the b ios in tes is de a c id osg ra sos that are es tru c tu ra lm in te d ife re n te s ( For the most part, in the long term, the number of branches of the population, etc.) to those of the native age of the native region. These people have been able to see the logo so that they can be seen so that they do not look at each other so they can be seen in the same way. This is the region in which the federal government is responsible for the commission, and as the leader of the government of the French Republic. to come rc ia l desired. P ore jem p lo, the enzymes FA S of Bacillus subtilis, Saccharomyces cerevisiae, Streptomyces spp., Ralstonia, Rhodococcus, Corynebacteria, Brevibacteria, Mycobacteria, vad u ras or lea g in osa s, os im ila res, can expire in a suitable host. The expiration of days in z ex as ex o ge na sa lte ra ra the structure of the id og ra so p roduc tion.

C u an do un h o sp e d a d o r de p roducc ion se g e n o m a n ip u la para p ro d u c ir un ac ido g raso con un n ive l de insa tu rac ion , ram ificac io n , o lo ng itud de la cad en a de ca rbono , esp ec ificos , el ac ido g raso g e n o m a n ip u la d o resu ltan te puede u tiliza rse en la p ro du cc ion de de riva d o s de ac ido s g rasos. Los d e riva dos de ac ido s g ra sos g e ne rado s a pa rtir de d icho s h o sp e d a d o re s de p ro du cc ion pueden m os tra r las ca ra c te ris tica s de l a c id o g ra so ge no m a n ip u la d o .

When a person is at the age of production, they are generated for a general purpose with a level of income, rationale, branching, or the extent of the body. In the area of carbon, especially the genoman ip ac i u u resu ltan can be used in the p r du du rion of de riva dos de arc sos rasos. Those from Riva Dos de Ac Ido Sg Ra are a member of the Department of Social Security, they can show you the most critical aspects of the situation. so ge no man ip u la do.

P or e jem p lo , un h o sp e d a d o r de p ro du cc ion pued e g e n o m a n ip u la rs e pa ra c re a r a c id o s g ra sos ra m ifica d o s de cad en a corta , que de spu es el h o sp e d a d o r de p ro du cc ion puede u tiliza r para p ro d u c ir a lcoh o le s g ra sos ra m ifica d o s de ca d e n a corta . De m an e ra s im ila r, g e n o m a n ip u la n d o un h o sp e d a d o r de p ro du cc ion pued e p ro du c irse un h id ro ca rbu ro para p ro d u c ir un ac ido g raso que te n g a un n ive l de ram ificac io n , in sa tu ra c ion , y /o lo ng itud de la cad en a de carbono de fin id o s y p ro du c ien do a s i una po b lac ion de h id ro ca rbu ros hom og ene a . P ara m e jo ra r la h o m og ene id ad del p ro du c to resu ltan te pueden em p le a rse e ta pas ad ic io na le s . P o r e jem p lo , cu a n d o se desea un a lcoh o l in sa tu ra do , un e s te r g ra so o un h id ro ca rbu ro , el o rg a n ism o h o s p e d a d o r de p ro d u cc io n puede g e n o m a n ip u la rse pa ra p ro d u c ir n ive le s ba jos de a c id o s g ra sos sa tu ra do s , y a d e m a s pued e m o d ifica rse para e xp re sa r una d e sa tu ra sa ad ic io n a l y as i d is m in u ir la p ro du cc ion de p ro du c to sa tu rado .

P o e jem p lo, a ho p age of p ro du cc ion can egenoman ip u la rs e para ra c re ar a c id osg ra are the m icants of cad in short, which spu is the Now, the age of P ro du cc ion can be used to go to Lcoh or continue to be short-lived. In my opinion, genoman ip u la ndo a ho sp age of p ro du cc ion can ep ro du c go a h id ro ca rbu ro for p ro duc go a satin ac i g satin that you There is a branching level, in sa tu ra tion, and / or the end of the carbon in the end carbon, as well as a small number of people. ro ca rbu ros hom og ene a. P o m e jo ra r the h o m og ene id ad of the p ro du c to resu ltan can be p e r e to be added to the pas ad ic io na le s. P o jem p lo, when you want a lcoh ol in sa tu ra do, a e s rg ra so or a h id ro ca rbu ro, the o rg an ism ohosoder of p ro du cc io n can genoman ip u la rse pa ra p ro duc go n ive le s jjos de a c id osg ra sos sa tu ra do s, also can be emod ified to e xp re sa r one of sa tu ra sa ad This is the way to go for your time.

A. Grupos ramificados y ciclicos

A. Branched and cyclic groups

1. G e n o m a n ip u la c io n de d e riva d o s de ac ido s g ra sos ra m ifica d o s y c ic lico s

1. G e n o m a n ip u la c e n d e riva d o s de ac ac s g ra sos ra m ifica d o s y ci lico s

Los ac ido s g ra sos son p ro du c to s in te rm e d io s c lave en la p ro du cc ion de d e riva d o s de ac ido s g rasos. U tilizando a c id o s g ra sos ram ificado s o c ic lico s pueden p re p a ra rse de riva dos de ac ido s g ra sos que con teng an pun tos de ram ificac io n , g ru po s c ic lico s y co m b in a c io n e s de los m ism os.

S s s sos p son son son son son son son son son son son son son son son son son la la la en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en en By using the identity of the sociological branch, they can be considered as a source of credit if they have branching points, such as the syllabus and co mb. in acions of the same.

P o r e jem p lo , E. coli p ro du ce de m an e ra na tu ra l ac ido s g ra sos de cad en a linea l (aFA s, de l ing les straigt chain fatty acids). P ara g e n o m a n ip u la r E. coli pa ra p ro d u c ir ac idos g ra sos de cadena ram ificada (b rFA , del ing les branched chain fatty acids), va r io s genes que p ro po rc ion an p re cu rso res ram ificado s (p o r e jem p lo , un operon bkd ) pueden in tro d u c irse en el h o s p e d a d o r de p ro d u cc io n y e xp re sa rse pa ra p e rm itir el in ic io de la b io s in te s is de a c id o s g ra so s a p a rtir de p re c u rs o re s ra m ifica d o s (p o r e jem p lo , fabH). Las fa m ilia s de ge ne s bkd, ilv, icm y fab pu ed en e xp re sa rse o s o b re e xp re sa rse para p ro d u c ir d e riva dos de ac idos g ra sos de cad en a ram ificada . De m an era s im ila r, para p ro du c ir ac ido s g ra sos c ic licos, los genes que p ro po rc ion an p re cu rso res c ic lico s pueden in tro d u c irse en el h o sp e d a d o r de p ro d u cc io n y e xp re sa rse pa ra p e rm itir el in ic io de la b io s in te s is de ac ido s g ra sos a p a rtir de p re cu rso re s c ic licos . Las fa m ilia s de ge ne s ans, chc y plm pueden e xp re sa rse o s o b re e xp re sa rse para p ro d u c ir ac ido s g ra sos cic licos.

P o jem p lo, E. coli p ro du ce de m an e ra na tu ra l ac l se s ra ra es de cad en a line l (aFA s, from l ing les straigt chain fatty acids). For the genoman ip u r E. coli for ra duc go ra raid acids (branded chain fatty acids), there are genes that allow for an Why do you want to get there (for example, a bkd operon) can enter the host of the host for the first time to begin the beginning of the year? However, it is also a matter of interest in the case (for example, fabH). The families of ge ne s bkd, ilv, icm and fab pu ed in e xp re sa rse osob re e xp re sa leave for p ro duc go de riva two of acidic g ra sos de cad in a ram ified In other words, for the sake of being cyclic, the genes that can be used by the cyclist can go into the home today. In the age of the public, it is necessary to begin the inception of the law, but it is to be separated from the physicists. The families of ge ans ans, chc and plm can e xp re sa rse osob re e xp re sa rse so that they can go cyclical.

A d ic io n a lm e n te , un h o sp e d a d o r de p ro du cc ion puede g e n o m a n ip u la rse para e xp re sa r genes que cod ifiq uen p ro te in a s para el a la rg a m ie n to de los b rF A (p o r e jem p lo , genes que co d ifica n la A C P , FabF, e tc .) y /o d e le c io n a r o a te n u a r los co rre sp o n d ie n te s genes de E. coli que n o rm a lm e n te con du cen a sFA s. En este sen tido , genes e n d o g e n o s que co m p e tiria n con los ge ne s in tro d u c id o s (p o r e jem p lo , fabH,fabF) se d e le c io n a n o a tenuan .

On a daily basis, a person of legal age may be able to use the genes coding if they are available for him or her. of the B rF A (for example, genes that codify the ACP, FabF, e tc.) and / or of the narrative, the numbers of the E. coli genes that No rm a lm in te con du cen a sFA s. In this sense, end genes or genes that would interact with the genes in tro ducts (for example, fabH, fabF) are known as nano nano.

La a c il-C o A ram ificada (p o r e jem p lo , 2 -m e til-b u tir il-C o A , iso va le ril-C o A , iso bu tir il-C o A , e tc .) son los p re cu rso re s de brFA . En la m ayo ria de los m ic ro o rg a n ism o s que con tien en brFA, los b rF A se s in te tizan en dos e ta pas a pa rtir de a m in o a c id o s ra m ifica d o s (p o r e jem p lo , iso leu c in a , le uc in a o v a lin a ) (K ad en a , M icrob io l. R ev., 55 :288 , 1991). Un h o s p e d a d o r de p ro du cc ion puede g e n o m a n ip u la rse para e x p re s a r o so b re e x p re s a r una o m as de las e n z im a s im p lica d a s en estas dos e ta pas para p ro d u c ir brFA s, o para s o b re p ro d u c ir los brFA. P o r e jem p lo , el h o sp e d a d o r de p ro du cc ion puede te n e r una en z im a e n d o ge na que pueda re a liza r una e tapa que con du zca un brFA , po r lo tan to , so lo los ge ne s que cod ifica n las e n z im a s im p lica da s en la seg u n d a e tapa deben in tro d u c irse de fo rm a reco m b in an te . Las t io e s te ra s a s m u ta n te s de la d ivu lg ac ion pueden g e n o m a n ip u la rse pa ra te n e r una o m as p ro p ieda de s a lte radas, po r e jem p lo , e sp e c ific id a d a lte ra d a y /o ac tiv ida d (p o r e jem p lo , ve lo c id a d ca ta litica ) au m en ta da , con resp ec to a los co m p u e s to s de a c il-C o A o a c il-A C P de cad en a ram ificada o c ic lica de sc rito s en el p re sen te do cu m e n to . P or co n s ig u ie n te , pueden fa b rica rse ce lu la s re co m b in a n te s p ro d u c to ra s de d e riva d o s de ac ido s g ra sos para p ro d u c ir p re fe re n te m e n te un p ro d u c to de riva d o de ac ido s g ra sos d e se a d o de ca d e n a ra m ifica d a o c ic lica que pued e te n e r un a lto v a lo r co m o p ro du c to fina l.

Acid C-C or A ram ified (for example, 2-me til-b u tir il-C or A, iso va le ril-C or A, iso bu tir il-C or A, e tc. ) are the most important of brFA. In the majority of the countries that are in brFA, the B rF A is divided into two stages to start from am in or to the rationale. (by jem p lo, iso leu c in a, le uc in aova lin a) (K ad en a, M icrob io l. R ev., 55: 288, 1991). A host of public relations may generate ip to use one or more of the enzymes implied in these two stages for this purpose, or to go over the brFA. P o jem p lo, the ho p age of p ro du cc ion can have a cover that can make a cover with a brFA, so much so, so The genes coding for the implicit enzymes in the second cover must be introduced from one to another. The uncles of the d ictu lg ac ion can genoman ip u la rse pa to have an om rs p ro p ieda de sa lte radas, po re jem p lo, e sp ec ific id ada lte ra day / o ac tiv ida d (pore jem p lo, see c id ad ca ta litica) au m en ta da, with respect to the c acts of ac il-C or A oac il-A CP of cad in the oc clic branched of sc rito s in the p re sen te do cu men to. For example, they may be able to see what they are doing in the case of de riva dos de acido sg ra sos for p ro duc go p re faith re In this case, it is only a matter of interest in the chain of information that can be used for a long period of time.

2. F o rm ac ion de ac ido s g ra sos ra m ifica d o s y d e riva d o s de ac ido s g ra sos ram ificado s

2. F o rm ac ation of s g ra sos ra m ifica d o s y d e riva d o s of ac i s g ra sos bran ified

La p rim era e ta pa en la fo rm a c io n de b rF A es la p ro du cc ion de los a -c e to a c id o s c o rre sp o n d ie n te s m ed ia n te una a m in o a c id o a m in o tra n s fe ra sa de cad en a ram ificada . Los ho sp e d a d o re s de p ro du cc ion pueden in c lu ir e n d o g e n a m e n te g e n e s que co d ifica n d icha s e n z im a s, o de m an e ra a lte rna tiva , d ich o s g e n e s pu ed en in tro d u c irse de m an e ra reco m b in an te . E. coli, po r e jem p lo , exp resa de fo rm a e n do ge na d icha enz im a , IlvE (E C 2.6.1.42 ; n.° de reg is tro Y P _ 026247 de G e n B a n k ). En a lg u n o s h o sp e d a d o re s de p ro du cc ion , una a m in o tra n s fe ra s a de a m in o a c id o s h e te ro loga de cadena ram ificada no puede exp resa rse . S in em bargo , si la enz im a IlvE de E. coli o cu a lq u ie r otra a m in o tra n s fe ra sa de am in o a c id o s de cadena ram ificada (p o r e jem p lo , IlvE de Lactococcus lactis (n .° de reg is tro A A F 34406 de G en B ank), IlvE de Pseudomonas putida (n .° de reg is tro de G e n B a n k N P _ 745648 ) o IlvE de Streptomyces co e lico lo r(N u m e ro de reg is tro N P _ 629657 de G en B ank)), no es endo ge na , puede in tro du c irse . Si la reacc ion de la a m in o tra n s fe ra sa es lim itan te de la v e lo c id a d en la b ios in tes is de b rF A en el o rg an ism o h o sp e d a d o r de p ro du cc ion se lecc io na do , en to n ce s la a m in o tra n s fe ra sa pued e so b ree xp resa rse .

The p rim era e ta pa in the fo rm ac io n of b rF A is the p r du du tion of the a osce r a sp id rre sp ond ie n te sm ed ia n te an am in o a c id oam in o tra ns fe ra sa de cad in branched branch. The hoped e o r ents of p ro du cc ion can in c lu go end o genamen te genes that codifies nd icha senz im as, or from m an e ra ra lt rna tiva, d ich osgenes pu ed en in tro duc leave m an e ra ra mb in an te. E. coli, po re jem p lo, exp resa de fo rm aen do ge na d icha enzy im a, IlvE (EC 2.6.1.42; reg. No. YP _ 026247 of G in B ank). In some recent years of protection, an amne or other faiths of am in or in which the branched-chain logotype cannot expire. However, if the enzyme is associated with E. coli or when it is another year in the process of branching or branched chain (for example, lvE, IlvE) of Lactococcus lactis (Reg. No. AAF 34406 of G in B ank), IlvE of Pseudomonas putida (Reg. No. of G in B ank NP _ 745648) or IlvE of Streptomyces co lico lo r (Registered number NP _ 629657 of G in B ank)), it is not endogenous, it can be introduced. If the reaction of the am in or tra ns faith ra sa is limited to the truth in the b ios ies tes of b rF A in the o rg an ism oho sp age of p ro du cc The lesson was read in all the years in which faith can be carried out.

La seg un da e tapa es la d e sca rb o x ila c io n ox id a tiva de los a -c e to a c id o s a la c o rre sp o n d ie n te a c il-C o A de cadena ram ificada . E s ta rea cc io n pued e c a ta liza rla un c o m p le jo de a -ce to ac ido d e s h id ro g e n a s a de ca d e n a ra m ifica d a (bkd; EC 1.2.4.4 ). (D e no ya et al., J. B acte rio l., 177: 3504, 1995), que con s ta de las su b u n id a d e s E 1 a /p (d e sca rb ox ilasa ), E2 (d ih id ro lip o il tra n s a c ila s a ) y E3 (d ih id ro lip o il de sh id ro g e n a sa ). E stos co m p le jo s de a -ce to ac ido d e sh id ro g e n a sa de cad en a ram ificada son s im ila re s a los com p le jos de p iruva to y a -c e to g lu ta ra to d e sh id rog en asa . C ada m ic ro o rg a n ism o que posee los b rF A y /o que crece en am in o a c id o s de cad en a ram ificada , puede u tiliza rse com o fue n te pa ra a is la r genes bkd para la exp res ion en ho sp e d a d o re s de p ro du cc ion ta le s com o , po r e jem p lo , E. coli. A s im ism o , E. coli t ie n e el c o m p o n e n te E3 com o pa rte de su co m p le jo p iruva to d e sh id ro g e n a sa (cod ifica do , po r e jem p lo , po r lpd, (E C 1.8.1.4 ), n.° de reg is tro N P _ 414658 de G en B ank), po r lo tan to , puede s e r su fic ie n te para e x p re s a r so lo los genes E1 a /p y E2 bkd. La T ab la 2 en u m e ra e jem p los no lim itan te s de ge ne s bkdde va rios m ic ro o rg a n ism o s que pueden in tro d u c irse de fo rm a re co m b in a n te y exp resa rse en un h o sp e d a d o r de p roducc ion pa ra p ro p o rc io n a r p re cu rso re s de a c il-C o A de cad en a ram ificada . C o m o ho sp e d a d o re s de p ro du cc ion tam b ien pu ed en u tiliza rse m ic ro o rg a n ism o s que tie n e n d icho s ge ne s bkd.

The second day is that of sca rb ox ila c io n ox id a tive of the a-c to c c ive id la cor rre sp ond ie n te ac il-C or A of branched chain. This can be done by a compliant person who has not been able to obtain a large amount of money (bkd; EC 1.2.4.4). (D e no ya et al., J. B acte rio l., 177: 3504, 1995), which with s of its bun id ades E 1 a / p (from sca rb ox ilasa), E2 (d ih id ro lip or il tra nsac ila sa) and E3 (d ih id ro lip or il de sh id ro gena sa). These co-laws are a-c to c gone from sh id ro gena sa de cad in a ram ified are s im ila re sa the com p le jos of p iruva to ya -ce to g lu ta ra to of sh id rog in handle. Each one that owns the B rF A and / or that grows in am in or in the ways of branching, can be used as it was for the r bkd genes for the expiration in ho sp e re r ents of p ro du cc ion ta le s com o, po re jem p lo, E. coli. Also, E. coli t ie ne the component E3 com o part of its co p le jo p iruva to de sh id ro gena sa (codified, po re jem p lo, po r lpd, (EC 1.8.1.4), reg.number NP _ 414658 of G in B ank), therefore, it may be sufficient to express only genes E1 a / p and E2 bkd T ab la 2 en ume ra e jem p not limited to ge ne s bkdde va rios m icro or rg an ismos that can in tro duc leave fo rm a re co mb in an te and exp Review a person at the age of production for the first time of ac-C or A of the branch in a branch. How do you expect to be able to use it as well as what you need to know ?

Tabla 2: Genes B k d de^ microor anismos seleccionados

Table 2: Genes B kd of ^ selected microorganisms

co n tin u a c io n

continuation

En otro e jem p lo , en un h o sp e d a d o r de p roducc ion , puede fa b rica rse iso bu tiril-C o A , po r e jem p lo en E. coli, a tra ve s de la coe xp re s io n de una c ro to n il-C o A red u c ta sa (C cr, EC 1.6.5.5 , 1.1.1.1 ) e is o b u tir il-C o A m u tasa (sub un ida d g ra nd e Icm A, EC 5.4.99.2 ; su b u n id a d pe quena Icm B, EC 5.4.99.2 ) (H an y R eyno lds , J. B acte rio l., 179 :5157 , 1997). La c ro to n il-C o A es un p ro d u c to in te rm e d io en la b io s in te s is de a c id o s g ra so s en E. coli y o tros m ic ro o rg a n ism o s . En la Tabla 3 se o frecen e je m p lo s no lim itan te s de genes ccr e icm de m ic ro o rg a n ism o s se lecc io na do s .

In another case, in an old age of production, it is possible to isolate C-A or A, as in E. coli, through the coefficient xp re s A network with an il-C or A red uc ta sa (C cr, EC 1.6.5.5, 1.1.1.1) and is required to pay a fee (under one way dg ra nd e Icm A, EC 5.4.99.2; its bun id ad pe quena Icm B, EC 5.4.99.2) (H an y R eyno lds, J. B acte rio l., 179: 5157, 1997). The c ro to n il-C o A is a p du du to in te rm ed io in the b io s in te s is de a c id osg ra so s in E. coli yo tros m ic ro o rg an ism os. In Table 3, they are not limited to the ccr and icm genes of m icro or rg an ism os le le io na do s.

r i mr i m

A d e m a s de la exp res ion de los genes bkd, el in ic io de la b ios in tes is de b rF A utiliza p -ce to a c il-p ro te in a s in ta sa III tra n s p o rta d o ra de ac ilo (FabH , EC 2.3.1.41 ) con e sp ec ific ida d para las a c il-C o A de cad en a ram ificada (L i et al., J. B ac te rio l., 187 :3795 -3799 , 2005). En la Tabla 4 se en um e ran e jem p los no lim itan te s de d ichas e n z im a s F a b H . Los genes fabH im p lica do s la b ios in tes is de ac ido s g rasos de cu a lq u ie r m ic ro o rg a n ism o que con teng a b rF A pueden exp re sa rse en un h o sp e d a d o r de p ro du cc ion . Las en z im a s Bkd y FabH de ho sp e d a d o re s de p ro du cc ion que no p ro du cen b rF A de m an era na tu ra l puede que no sop orten la p ro du cc ion de brFA, po r lo tan to , Bkd y FabH pueden exp re sa rse de fo rm a reco m b in an te . V e c to re s que con tienen los genes bkd y fabH pueden in se rta rse en d icho h o s p e d a d o r de p ro du cc ion . De m a n e ra s im ila r, el n ive l e n d o g e n o de la p ro du cc ion de Bkd y FabH pued e que no sea su fic ie n te para p ro d u c ir brFA, po r lo tan to , pueden sob re e xp re sa rse . A d ic io n a lm e n te , o tros co m p o n e n te s de la ruta de la b io s in te s is de ac ido s g ra sos pu ed en exp re sa rse o sob re e xp re sa rse , ta le s co m o las p ro te inas tra n sp o rta d o ra s de ac ilo (A C P ) y la p -ce to a c il-p ro te in a s in ta sa II tra n s p o rta d o ra de ac ilo (co d ifica d a po r fabf EC 2.3.1.41 ) (en la Tabla 4 se e n u m e ra n e je m p lo s no lim ita n te s de ca n d id a to s ). A d e m a s de e x p re s a r es to s genes, a lg u n o s ge ne s en la ru ta de la b ios in tes is de ac ido s g ra sos e n do ge nos pueden a te n u a rse en el h o s p e d a d o r de p roducc ion . Los genes que co d ifica n e n z im a s que c o m p ite n po r el su s tra to (o su s tra to s ) con las e n z im a s de la ru ta que dan co m o resu lta d o la p ro du cc ion de brFA, pueden a te n u a rse para a u m e n ta r la p ro du cc ion de brFA . P o r e jem p lo , en E. coli los c a n d id a to s que in te rfie ren m as p ro b a b le m e n te con la b ios in tes is de fabH son los genes fabH (n .° de reg is tro N P _ 415609 de G e n B a n k ) y /o fabF (n .° de reg is tro N P _ 415613 de G enB ank).

In addition to the expiration of the bkd genes , the beginning of the b ios tes is de b rF A uses p -ce to ac il-p ro teins as in ta sa III tra nspo rta do ra of acyl (FabH, EC 2.3.1.41) with e sp ec ific ida d for ac il-C or A of cad in a branched branch (L et al., J. B ac te rio l., 187 : 3795-3799, 2005). Table 4 shows the limitations of the enzymes F ab H. The fabH imp plicic genes of the b ios ies tes de ac a sg rass de cu a lq u ie rm icro or rg an ism or which with ten ab rF A can be expressed in a ho age of p ro du cc ion. Those in z b as Bkd and FabH of ho sp edado re s of p ro du cc ion that not p ro du cen b rF A of m an era na tu ra l may not support the p ro du cc ion of brFA Because of this, Bkd and FabH can express themselves from fo rm to reco mb in an te. V ec to re s that with the bkd and fabH genes can be inserted in that host host. In a similar way, the level of the Bkd and FabH p r du du tion may not be sufficient for brFA, but so far, they can also be re e xp re sa rse. A d ic io na lm en te, or others compose te s of the route of the b io s in te s is de ac ido sg ra sos pu ed in exp re sa rse or sob re e xp re sa rse, ta I know how to work with acyl groups (ACP) and I want to know how to work in the second place of acyl ( COD ification of the Fabf EC 2.3.1.41) (Table 4 lists the requirements not limited to each other). Other than this, it is all genes, and some people in the path of the bishops are also able to get in the host of the host ion production. The genes that encode nenz im so that comp ite n po r the his (or his s) treatment with the enzymes of the route that give as a result of the p r du du ion of brFA, they can be added to increase the brFA of brFA. P o jem p lo, in E. coli, the candidates who in r rie rie more than with the basics of fabH are the fabH genes (reg. NP _ 415609 of G in B ank) and / or fabF (reg. No. NP _ 415613 of G inB ank).

Tabla 4: fa b H ACP fa b F enes de microor anismos seleccionados con brFA

Table 4: fa b H ACP fa b Microorganisms selected with brFA

co n tin u a c io n

continuation

C o m o se ha m e n c io n a d o an te rio rm e n te , pueden p ro du c irse a lcoh o le s de cad en a ram ificada m ed ia n te la co m b in a c io n de genes de exp res ion que soportan la s in tes is de b rF A y la s in tes is de a lcoho l. P o r e jem p lo , cua nd o un gen que co d ifica una a lcoh o l reduc tasa , ta l com o acrl de Acinetobacter baylyi A D P 1 , se co e xp re sa con un operon bkd en una ce lu la h o spe da do ra de E. coli, la ce lu la h o spe da do ra puede s in te tiz a r iso pe n tan o l, iso bu ta no l o 2 -m e til bu tano l. De m a n e ra s im ila r, cua nd o el gen acrl se coe xp re sa con los genes ccr/icm en una ce lu la h o sp e d a d o ra de E. coli, la ce lu la h o sp e d a d o ra pued e s in te tiz a r isobu tano l.

How rm has been mentioned before you can go to Lcoh or read about it in the same way as you can find it in terms of export genes that they support the s is of b rF A and the s is of a lcoho l. P ore xam p I, CUA nd or a gene co ifica d one to lcoh ol reduc rate, ta l com or ACRL Acinetobacter baylyi ADP 1 co e xp re sa an operon bkd in an EC lu ho spe From E. coli, the city of the house can be used as an iso, but not quite useful. In a similar way , when the acrl gene is co- xp re sa with the ccr / icm genes in a city of E. coli, the city is in it is isobu tano l.

3. F o rm ac ion de ac ido s g ra sos c ic lico s y d e riva d o s de ac ido s g ra sos c ic licos

3. F o rm ac ation of s g ra sos c ci l lico s y d e riva d o s de acc s g ra sos cic licos

P ara tra n s fo rm a r un h o sp e d a d o r de p roducc ion , ta l com o una bacte ria E. coli, en un o rg a n ism o cap az de s in te tiza r ac ido s g ra sos w c ic lico s (c iFA ), un gen que p ro p o rc io n a la c ic lo h e x ilc a rb o n il-C o A (C H C -C o A ) p re cu rso ra c ic lica (C ro pp e t al., N a tu re B io tech ., 18: 980 -983 , 2000), se in tro du ce y se exp resa en el h o sp e d a d o r de p ro du cc ion . Una co n ve rs io n s im ila r es pos ib le pa ra o tros ho sp e d a d o re s de p ro du cc ion , po r e jem p lo , bacte rias , le vad u ras y hongos fila m en to sos .

For the first time a senior citizen, such as a bacteratia E. coli, in one of the most important countries in the world s (c iFA), a gene that p ro po rc io na the c iclo hex ilc a rb on il-C or A (CHC-C or A) for the cyclic ration (C ro pp et al ., N a tu re B io tech., 18: 980-983, 2000), is introduced and expired in the old age of the pipeline. A co n ve rs io ns im ila r is pos ib le pa ra o tros ho sp edado re s de p ro du cc ion, po re jem p lo, bacte rias, le va u u ras and fungi row m en to sos .

C o m o e je m p lo s no lim ita tivo s de ge ne s que p ro p o rc io n a n C H C -C o A en E. coli se inc luyen : ansJ, ansk, ansL, chcA y ansM de l g ru po de genes de a n sa trie n in a de Streptomyces collinus (C hen et al., Eur. J. B iochem ., 261: 98-107, 1999), o plmJ, plmK, plml, chcA y plmM del g ru po de genes de fo s la c to m ic in a B de Streptomyces sp. HK803 (P a la n ia p p a n et al., J. Biol. C hem ., 278: 35552 -35557 , 2003 ) ju n to con el gen chcb (P a tton et al., B iochem ., 39: 7595 -7604 , 2000 ) de S. collinus, S. avermitilis o S. coelicolor (vea se la Tabla 5 para los num eros de reg is tro de G en B ank). Los genes e n u m e ra d o s a n te rio rm e n te en Tabla 4 pueden exp resa rse de spu es para pe rm itir el in ic io y el a la rg a m ie n to de ac ido s g ra sos w c ic licos. C om o a lte rna tiva , los genes ho m o lo g o s pueden a is la rse de m ic ro o rg a n ism o s que p roducen cyF A y e xp re sa rse en E. coli.

How do you think it is not limited to people who want CHC-C or A in E. coli include: ansJ, ansk, ansL, chcA and ansM of lg ru po of genes from an sa trie n in a of Streptomyces collinus (C hen et al., Eur. J. B iochem., 261: 98-107, 1999), or plmJ, plmK, plml, chcA and plmM of the gene group de fo s la c to m ic in a B by Streptomyces sp. HK803 (P a la n ia ppan et al., J. Biol. C hem., 278: 35552-35557, 2003) ju with the chcb gene (P a tton et al., B iochem., 39: 7595 -7604, 2000) of S. collinus, S. avermitilis or S. coelicolor (see Table 5 for registration numbers of G in B ank). The genes listed in the table in Table 4 may expire from each other in order to allow the beginning of the period and the continuation of the situation. In accordance with this, the genes can be found in the same way as they produce cyF A and xp re sa rs in E. coli.

Los genes e n u m e ra d o s en la Tabla 4 (fabh, A C P y fabF) son su fic ie n te s pa ra p e rm itir el in ic io y el a la rg a m ie n to de ac ido s g ra sos w c ic lico s po rque no rm a lm e n te tien en una am p lia esp ec ific ida d de sustra to . Si la coe xp re s ion de a lgu no de estos genes con los genes ansJKLM/chcAB o pmlJKLM/chcAB de la Tabla 5 no p ro du ce cyF A , en tonces pueden a is la rse ho m o lo g o s fabH, ACP y /o fabF de m ic ro o rg a n ism o s que p roducen c yF A (p o r e jem p lo , u tilizando ce b a d o re s de P C R de g e n e ra d o s o son da s de se cu e n c ia s de A D N h e te ro lo g a s ) y pueden coe xp re sa rse . La Tabla 6 en u m e ra e je m p lo s no lim ita tivo s de m ic ro o rg a n ism o s que con tien en ac ido s g ra sos w cic licos.

The genes listed in Table 4 ( fabh, ACP and fabF) are the only way to start the process and the way of accruing it to the future. s po rque not rm a lm in te lie in an amp esp esp ific ida d de subtra to. If the coe xp re s ion of Some of these genes with the ansJKLM / chcAB or pmlJKLM / chcAB genes in Table 5 do not exist in cyF A, then they can be used as fabH, ACP and / or fabF of m ic R o rg an isms that produce c and F A (for example, by using gene-based PCR primers), it is only a matter of DNA DNA that has caused gas) and can xp re sa rse. Table 6 in umera e je mp lo s is not limited to m icro or rg an isms that have a cyclic ac i ct.

T l : E m l n limi iv mi r r ni m n i n n i r i li

T l: E m l n limi iv mi r r ni m n i n n i r i li

B. Saturacion

B. Saturation

Los ac ido s g ra sos son p ro du c to s in te rm e d io s c lave en la p ro d u cc io n de d e riva d o s de ac ido s g rasos. Los g ra do s de sa tu ra c io n en d e riva d o s de ac ido s g ra sos pueden con tro la rse re g u land o los g ra do s de sa tu ra c io n de los p roduc tos in te rm e d io s de ac idos grasos. Las fa m ilia s de genes sfa, gns y fab pueden e xp re sa rse o so b re e xp re sa rse para c o n tro la r la sa tu ra c io n de ac ido s grasos.

Sg ra are those that are safe in rve ed io sc lave in pro du cc io n de de riva two of ac cg sos rasos. The g ra do s de sa tu ra tio n in riva dos de ac se g se s ra ra s can be with the re rs re gu land or the g ra do s sa sa ra ra tio of the products in te te rm ed io s of fatty acids. The families of sfa, gns and fab genes can be taken to be able to get to know the fatty acids.

Los ho sp e d a d o re s de p ro du cc ion pueden g e n o m a n ip u la rse pa ra p ro d u c ir ac ido s g ra sos in sa tu ra dos g e n o m a n ip u la n d o el h o sp e d a d o r de p ro du cc ion para so b re e x p re s a r fabb o ha c iendo c re ce r el h o s p e d a d o r de p ro du cc ion a ba ja s te m p e ra tu ra s (p o r e jem p lo , m en os de 37 °C ). FabB tien e p re fe re n c ia p o r c is -53 de ce n o il-A C P , y da co m o re su lta d o la p ro du cc ion de ac ido s g ra sos in sa tu ra d o s en E. coli. La so b re e xp re s io n de l gen fabB da com o resu lta d o la p ro du cc ion de un p o rcen ta je s ig n ifica tivo de ac ido s g ra sos in sa tu ra d o s (de M endoza e t al., J. Biol. C hem ., 258 :2098 -101 , 1983). El gen fabB puede in se rta rse y exp resa rse en ho sp e d a d o re s de p ro du cc ion que no tie n e n el gen de fo rm a na tu ra l. E stos ac ido s g ra sos in sa tu ra d o s pueden u tiliza rse com o p ro du c to s in te rm e d io s en los h o sp e d a d o re s de p ro du cc ion que estan g e n o m a n ip u la d o s pa ra p ro d u c ir d e riva d o s de ac ido s g rasos, ta le s com o a lco h o le s g rasos, es te re s g rasos, ceras, o le finas, a lcanos, y s im ila res.

Ho sp eados re s de p ro du cc ion can genoman ip u la rse para p ro duc go ac c sg ra sos in sa tu ra two genoman ip u la ndo el ho sp ageor p p du du ion re exp o f re sar fabb or has re ce c r c iendo the host p ro du cc ion ja ba s mpe you ra ra s your (pore xam p I, m I 37 ° C). FabB has ep re fe re nc ia porc is -53 ce ce il il-A CP, and gives its status as the p r du du ction of ac i g sg ra sos in sa tu ra ra in E. coli. The so b re e xp re s io n of the gene fabB gives as a result of the p r du du tion of a poce ta s s significant ac tive of sg ra sos in sa tu ra ra ( M endoza et al., J. Biol. C hem., 258: 2098-101, 1983). The fabB gene can be inserted and expired in a residential state that does not have the foreign gene. These conditions can be used as part of the process in which the two countries that are gen ip and the two countries For the purpose of going from two to two years of age, it is common to those who are simple, this is true, waxes, or fine, to the hands, and the images.

C o m o a lte rna tiva , rep reso res de la b ios in tes is de ac ido s grasos, po r e jem p lo , un re p re so r (n.° de reg is tro N P _ 418398 de G e n B a n k ) co d ifica d o po r fabR pueden de le c io n a rse . E sto ta m b ie n da ra com o resu lta d o una m a yo r p ro du cc ion de ac ido s g ra sos in sa tu ra d o s en E. coli (Z han g e t al., J. Biol. C hem ., 277 :15558 , 2002 ). P ueden e fe c tu a rse d e le c io n e s s im ila re s en o tro s h o sp e d a d o re s de p ro d u cc io n . S e pu ede lo g ra r un a u m e n to a d ic io n a l en los ac ido s g ra sos insa tu ra dos , po r e jem p lo , po r so b re e xp re s io n de fabM (que cod ifica la trans -2 , c is -3 -d e ce n o il-A C P iso m erasa , n.° de reg is tro D A A 05501 de G e n B a n k ) y e xp re s io n co n tro la d a de fabk (qu e co d ifica la tra n s -2 -e n o il-A C P red u c ta sa II, n.° de reg is tro N P _ 357969 de G e n B a n k ) de Streptococcus pneumoniae (M a rra kch i et al., J. Biol. C hem ., 277: 44809 , 2002 ), m ie n tra s fab l se d e le c io n a de E. coli(que co d ific a la tra n s -2 -e n o il-A C P red uc tasa , n.° de reg is tro N P _ 415804 de G en B ank). A d ic io n a lm e n te , para a u m e n ta r el po rcen ta je de e s te re s g ra sos in sa tu ra dos , el h o s p e d a d o r de p ro du cc ion ta m b ie n puede so b re e x p re s a r fabB (que co d ifica la p -ce to a c il-A C P s in ta sa I, n.° de reg is tro B A A 16180 de G en B ank, (E C 2.3.1.41 ), sfa (qu e cod ifica un su p re so r de fabA n.° de reg is tro A A C 44390 de G en B ank), y gnsA y gnsB (am b os cod ifica n su p re so re s m u tan tes nu los secG, n.° de reg is tro A B D 18647.1 de G e n B a n k y n.° de reg is tro A A C 74076.1 de G e n B ank , resp e c tiva m e n te ). En a lg u n o s e jem p los, el gen e n d o g e n o fabF pued e a te nua rse , a u m e n ta n d o a s i el po rcen ta je de pa lm ito le a to (C ¹⁶.¹ ) p roduc ido .

C omoa lte rna tiva, representatives of b ios ies tes of fatty acids, po re jem p lo, a re p re so r (reg. No. NP _ 418398 of G en B ank ) The code can be read . This will also be the result of a ma r r th ro du ction of ac i g s s ra ra in sa tu ra ra in E. coli (Z have got al., J. Biol. C hem. , 277: 15558, 2002). They can be faith in le c io ness im ila re s en o tro sho sp edado re s de p ro du cc io n. It is possible to add an increase to the number in the following years, in which you are able to do so, because you are responsible for fabM (which codifies the trans -2, c is -3 -de ce non il-A CP iso m erasa, reg.number DAA 05501 of G in B ank) and xp re s io n with the fabk day ( What does the ns -2 -eno il-A CP red uc ta sa II, reg no. NP _ 357969 of G in B ank) code of Streptococcus pneumoniae (M a rra kch i et al. , J. Biol. C hem., 277: 44809, 2002), but is also known as E. coli ( which is also known as -2 -eno il-A CP) network uc rate, registration number NP _ 415804 of G in B ank). A d ic io na lm en te, in order to increase the power of these people, you will be able to keep them up, the host of this situation can also be able sar fabB (which codifies the p-ce to ac il-A CP s in ta sa I, reg.number BAA 16180 of G in B ank, (EC 2.3.1.41), sfa (qu e cod He writes his p re so r of fabA reg.number AAC 44390 of G in B ank), and gnsA and gnsB (both codify his p re so re smu tan so nu the secG, n. ° reg is tro ABD 18647.1 G in B anky n. ° is reg tro AAC 74076.1 G in B ank, resp ec tive ly). in lg unose xam p those, the gene end or geno FABF pued ea It has been added to you, the increase in power of the country is increased (C ^{16. 1} ).

Las t io e s te ra sa s m u tan tes de la d ivu lg ac ion pueden g e n o m a n ip u la rse para te n e r p ro p ieda de s a lte radas , por e jem p lo , e sp e c ific id a d a lte rad a y /o ac tiv idad a u m en ta da , con resp ec to a los co m p u e s to s de a c il-C o A o ac il-A C P su s titu id o s o no su s titu id o s que se p re pa ran com o se de sc rib e en el p re se n te do cu m e n to . P o r co n s ig u ie n te , la ce lu la re co m b in a n te p ro d u c to ra de d e riva d o s de ac ido s g ra sos pued e fa b rica rse para p ro d u c ir p re fe re n te m e n te un pe rfil de sa tu ra c io n d e se a d o en un p ro du c to de riva d o de ac ido s g ra sos que puede te n e r un a lto v a lo r com o p ro du c to fina l. C. Longitudes de cadena y caracteristicas de ester

The guys who are the leaders of the group can make a great deal of money to get to you, for the most part, and if you want to find the right rad / or current activity, with respect to ac c-C or A or ac il-A CP costs, its status is not its constituents that are considered common. or I know how to talk about it. For the same reason, the city will be the source of the riva two of the city, but it may be possible to go to the country to go to faith. Do not mention a profile of your life in a private sector, as you may find that a year is going to take place as soon as possible. . C. Chain lengths and ester characteristics

1. L o ng itud es de cad en a y p ro du cc ion de ca d e n a s im p ares

1. L o ng itud is of cad en a y p ro du cc ion of ca d e n a s im p ares

Los m e tod os d e sc rito s en el p re sen te d o cu m e n to pe rm iten la p ro du cc ion de e s te re s g ra sos y de riva dos de ac idos g ra sos de d ife re n te s lo ng itude s de cad en a s e le cc io n a n d o una t io e s te ra s a m u tan te ad ecu ad a que te n g a esp ec ific ida d y /o se le c tiv id a d po r un sus tra to de una lo ng itud de cad en a de ca rb o n o esp ec ifica . E xp re sa n d o las tio e s te ra sa s esp ec ificas , se pueden p ro d u c ir ac ido s g ra sos y de riva dos de ac ido s g ra sos que tien en lo ng itude s de cad en a de ca rb o n o d e sea da s . En a lg u n o s asp ec tos , una t io e s te ra s a e n d o g e n a pued e m u ta rse u tiliza n d o te c n ic a s de a lte rac ion g e n o m ica con oc idas . O, un gen que cod ifica una t io e s te ra s a p a rticu la r puede in tro d u c irse de m an e ra h e te ro loga en un h o s p e d a d o r de p ro du cc ion de ta l m an era que se p roduzca un ac ido graso o un de riva do de ac ido g raso de una lo ng itud de cad en a de ca rb on o pa rticu la r. En d e te rm in a d o s aspec tos , se su p rim e la exp res ion de t io e s te ra sa s en do g e n a s . Las t io e s te ra s a s m u ta n te s de la d ivu lg a c io n pu ed en g e n o m a n ip u la rse para te n e r p ro p ie d a d e s a lte radas , po r e jem p lo , e sp e c ific id a d a lte ra d a y /o ac tiv ida d au m en ta da , con re sp e c to a las lo n g itu d e s de ca d e n a e sp e c ifica s de los co m p u e s to s a c il-C o A o ac il-A C P de sc rito s en el p resen te do cu m e n to . P o r con s ig u ie n te , la ce lu la reco m b in an te que p ro du ce los de riva dos de ac ido s g ra sos puede fa b rica rse pa ra p ro d u c ir p re fe re n te m e n te un p ro du c to de riva do de ac ido s g ra sos con la lo ng itud de cad en a de se a d a y /o con un v a lo r a lto com o p ro du c to fina l.

The methods of sc rite s in the p re sen te do cu men to pe rm iten the p r du du cion of he re sg ra and two of acidic acids of d ife re I don't know what is happening in this way when you have an uncle, it will be so important that you will be interested in going through your work. of a ng itud of cad in a ca rb ono esp ec ifica. E xp re sa ndo uncle es te ra sa s esp ec ificas, you can p ro duc go ac se sg ra sos and de riva dos de acido sg ra sos you have in the ng itude s of cad in a de ca rb onode be da s. In some cases, an uncle is able to learn or use techniques from the genetic rationale with his eyes. Or, a gene that encodes an uncle is te sa sa rticu la r can in tro duc leave m an e ra he lo r lo lo lo in a host of the pipeline of this kind was to produce a fatty acid or a private source of a private matter in Canada or in Argentina. In terms of specific aspects, it is his uncle's expression as te r ra es s in do genas. The uncles are very common in the iv lg ac io n pu ed in genoman ip u la rse for te nerp ro p ie dadesa lte radas, po re jem p lo, e sp ec ific id ada lte ra day / o ac tiv ida d au m in ta da, with re sp ec to the ng itu des de ca denae sp e cific s of the com cues to sac il-C or A or ac il-A CP of sc rite s in the pasture. For example, the letter is recognized by the people of Riva dos de Acdo Sg Ra SOS. It can be manufactured to continue to go to faith. I mention you a little bit of money in the future, with the loyalty of Cad in a day and with a value of fine money.

En un aspecto , el de riva do de ac ido g raso con tiene una cadena de ca rb o n o de a p ro x im a d a m e n te 4 a 36 a to m o s de ca rb on o , de a p ro x im a d a m e n te 6 a 32 a to m o s de carbono , de a p ro x im a d a m e n te 10 a 30 a to m o s de ca rbono , de a p ro x im a d a m e n te 10 a 18 a to m o s de carbono , de a p ro x im a d a m e n te 24 a 32 a to m o s de carbono , de a p ro x im a d a m e n te 26 a 30 a to m o s de carbono , de a p ro x im a d a m e n te 26 a 32 a to m o s de carbono , de a p ro x im a d a m e n te 5 a 10 a to m o s de ca rbono , de a p ro x im a d a m e n te 10 a 16 a to m o s de c a rb o n o o de a p ro x im a d a m e n te 12 a 18 a to m o s de carbono . En un asp ec to a lte rna tivo , el de riva do de ac ido g raso con tiene una cad en a de ca rb on o m e n o r de a p ro x im a d a m e n te 20 a to m o s de ca rbono , m e n o r de a p ro x im a d a m e n te 18 a tom os de ca rbono , o m e n o r de a p ro x im a d a m e n te 16 a tom os de carbono . En otro aspecto , el p ro du c to de e s te r g raso es un p ro d u c to de e s te r g ra so sa tu ra d o o in sa tu ra d o que tie n e un co n te n id o de a to m o s de ca rb o n o en tre 24 y 46 a to m o s de ca rbono . En un aspecto , el p roduc to de e s te r g raso tien e un co n ten ido de a tom os de ca rb on o en tre 24 y 32 a to m o s de ca rbono . En o tro aspec to , el p ro d u c to de e s te r g ra so t ie n e un co n te n id o de ca rb o n o de 14 y 20 ca rb on os . En o tro aspec to , el e s te r g ra so es el e s te r m etilico de C ^{1 8 1}. En o tro aspecto , el e s te r g ra so es el e s te r e tilico de Cm-i. En o tro aspec to , el e s te r g ra so es el e s te r m e tilico de Cm-i. En o tro a sp e c to m as, el e s te r g ra so es oc ta d e c il e s te r de octano l.

In one aspect, the private sector with a chain of appraisal from 4 to 36 years old, of apprentices 6 to 32 at carbon level, from 10 to 30 minutes from carbon, from 10 to 18 carbon, from size 24 to 32 at carbon, from ap ro x im adamen te 26 to 30 to carbon, from ap ro x im adamen te 26 to 32 to carbon, from ap ro x im adamen te 5 to 10 amos carbon, from ap ro x im adamen te 10 a 16 to ca rb onoo from ap ro x im adamen te 12 to 18 a carbon. In an environmental context, the private sector with a private body has an appoor of less than 20 years, less than a year. im adamen te 18 a toms de ca rono, apor x ap ada x im adamen te 16 a tomos de carbon. In another aspect, the property of your country is a property of your family so that you have a relationship that has a common sense of interest. We can do it between 24 and 46 years ago. In one aspect, the product of the latter has a common sense of pipeline or between 24 and 32 years of carbon. In other respects, the pipeline of the latter will only have a channel of 14 and 20 years. In other respect, this is the ethyl ether of C ^{1 8 1} . In another aspect, this is the only one of Cm-i. In other respect, he is the only one in Cm-i. In other cases, this is only the case of octane.

C ie rtos m ic ro o rg a n ism o s p roducen p re fe re n te m e n te ac ido s g ra sos y de riva dos de ac ido s g ra sos de cad en a de ca rb o n o pa r o im par. P o r e jem p lo , E. coli n o rm a lm e n te p roduce ac ido s g ra sos y es te res e tilico s de ac ido s g rasos (F A E E ) de cad en a de ca rb on o par. S o rp re n d e n te m e n te , los m e to d o s de sve la d o s en el p resen te d o cu m e n to pueden u tiliza rse para a lte ra r esa p ro du cc ion . P o r e jem p lo , en c ie rta s c irc u n s ta n c ia s puede c re a rse una E. coli pa ra p ro d u c ir ac ido s g ra sos y F A E E de cad en a de ca rb o n o im par.

C ie rtos m icro o rg an ism osp roducen p re fe re n te men te ac se sg ra sos y de riva dos de ac a sg ra sos de cad in a ca rb ono para ro im par par. P o jem p lo, E. coli no rm a lm in te p roduce ac cido se s ra ra sos y es te te t e s tiles s de ca sos rasos (FAEE) de cad in a ca rb on o par. If you mention, the two methods of the two in the present can be used for the purpose of this protection. P o jem p lo, in that case it can be an E. coli to go du sc ra sg ra sos and FAEE de cad in a ca rb ono im par.

2. C a ra c te ris tica s de e s te r

2. C a ra c te ris tica s de e s te r

Un e s te r in c luye n o rm a lm e n te lo pu e d e d e n o m in a rs e co m o un lado "A " y un lado "B ". El lado B pu ed e c o n trib u ir con un ac ido g ra so p ro du c ido a p a rtir de s in te s is de novo en el o rg a n ism o h o sp e d a d o r de p ro du cc ion . En a lgunos asp ec tos , en los que el h o s p e d a d o r de p ro du cc ion se g e n o m a n ip u la a d ic io n a lm e n te pa ra p re p a ra r a lcoho les, in c lu ye n d o a lcoh o le s g rasos, el lado A ta m b ie n es p ro du c ido po r el o rg a n is m o h o s p e d a d o r de p ro du cc ion . En o tros a sp e c to s m as, el lado A puede s e r p ro p o rc io n a d o po r el m ed io de cu ltivo . S e le cc io n a n d o los genes de t io e s te ra sa desea do s , el lado B (y el lado A cuando se fa b rica n a lcoh o le s g ra sos ) puede d isen a rse para te n e r c ie rtas ca ra c te ris tica s d e sea b le s de la cad en a de ca rbono . E stas ca ra c te ris tica s inc luyen , po r e jem p lo , pun tos de ram ificac io n , pun tos de in sa tu ra c io n y lo ng itude s de cad en a de ca rb o n o de sea da s . P o r ta n to , las t io e s te ra sa s m u tan tes de la d ivu lg ac ion pueden g e n o m a n ip u la rse para te n e r p ro p ieda de s a lte radas , po r e jem p lo , esp ec ific ida d a lte ra d a y /o ac tiv idad au m en ta da , con resp ec to a la p re fe ren c ia po r a c e p ta r c ie rto s co m p u e s to s de a c il-C o A o ac il-A C P com o una cad en a la te ra l A com o se de sc rib e en el p resen te do cu m e n to . P o r con s ig u ie n te , la ce lu la re co m b in a n te p ro d u c to ra de d e riva d o s de ac ido s g ra sos pued e fa b rica rse de m an e ra que p ro d u zca p re fe re n te m e n te un p ro du c to de riva d o de ac ido s g ra sos d e se a d o que sea v a lio s o com o p ro du c to fina l.

An e s te r in c luye does not rm a lm in what I can edenden in a rs e as a side "A" and a side "B". The B side of the economy can go with a great deal of interest if it is de novo in the age of the age of age. In some cases, in which the host of the pipeline is generated by the addition of the law, in order to keep it safe, in turn. , the side A ta mb ie n is p ro du c gone by r o rg an is pho duyer of p ro du cc ion. In other cases, side A can be supported by the quantitative environment. If the genes of uncle are desired, the B-side (and the A-side when manufactured or not) can be designed to help you Ca ra c ta r t tica sde sea b le s of the cad in a ca rono. These are the most important of all, they are important, the points of in tu, sa pun pun pun pun pun pun pun pun pun cad cad cad cad cad cad cad cad cad cad cad cad you give . P o ta n t, the un t esio ra te smu tan tees of the d ivu lg ac ion can genoman ip u la rse for te nerp ro p ieda de sa lte radas, po re jem p lo, esp ec ific ida This is the day / o ac tiv idad au m en ta da, with respect to the p r r faith p racep ta rc ie rto s cocomues to ac il-C or A or ac il-A CP com or a cad in the te ra l A com o se sc sc e in the present. For the same reason, the city is the source of de riva dos de riva dos de sg ra so you can make a living out of me that you can zca p re faith re n te ment a p ro du c t riva do de r a c A ³ n sg ra he s of se lt be va lio so com op ro du c to fina l.

C u a n d o se se le cc io n a n ge ne s de t io e s te ra s a pa rticu la res , los lados A y B te n d ra n ca ra c te ris tica s de cad en a de c a rb o n o s im ila re s cuando am b os sean a p o rtad os po r el h o sp e d a d o r de p roducc ion u tilizan do p ro du c to s in te rm ed ios de la v ia b ios in te tica de ac ido s g rasos. P o r e jem p lo , al m enos a p ro x im a d a m e n te el 50 %, 60 %, 70 % o u 80 % de los es te res g ra sos p ro du c ido s te n d ra lados A y B cuya lo ng itud va rie en a p ro x im a d a m e n te 2, 4, 6, 8, 10, 12 o 14 ca rb on os . El lado A y el lado B ta m b ie n pu ed en m o s tra r n ive les de ram ificac io n y sa tu ra c io n s im ila res.

When you are told about this type of information, the sides A and B will make you laugh at the cadres of cars in each case. s when they are supported by the ho o age of p uduction or use of the duo in the r o bios of the via b inos in te tica of ac A ³ A A A A A A A A AƒAA‚AA‚AA‚A‚A‚AA‚A‚A‚A‚A‚A‚A‚A¡ P o jem p lo, at least about 50%, 60%, 70% or 80% of these, you will have sides A and B whose ng itud laughs in ap ro x im adamen te 2, 4, 6, 8, 10, 12 or 14 ca rb on os. The A side and the B side are also capable of branching levels and are your images.

A d e m a s de p ro d u c ir a lcoh o le s g ra sos a p o rtad os al lado A, el h o sp e d a d o r de p ro du cc ion puede p ro d u c ir o tros a lco h o le s de cad en a co rta ta le s co m o e tano l, p ropano l, iso p ro pan o l, Isob u tano l y bu tano l para su in co rp o ra c io n en el lado A u tiliza n d o te c n ic a s b ien c o n o c id a s en la m a te ria . P o r e jem p lo , el o rg a n ism o h o sp e d a d o r de p ro d u cc io n puede p ro d u c ir bu tano l. P ara c re a r ce lu las p ro d u c to ra s de bu tano l, la cepa LS 9001, po r e jem p lo , puede g e n o m a n ip u la rse a d ic io n a lm e n te pa ra e xp re sa r atoB (a ce til-C o A a ce tiltra n s fe ra sa ) de Escherichia coli K12, p -h id ro x ib u tir il-C o A d e sh id ro g e n a sa de Butyrivibrio fibrisolvens, c ro to n a sa de Clostridium beijerinckii, bu tir il C o A d e sh id ro g e n a sa de Clostridium beijerinckii, a ld e h id o d e sh id ro g e n a sa (A L D H ) de ac ilac ion de C o A de Cladosporium fulvum y adhE que co d ifica una a ld e h id o a lco h o l d e s h id ro g e n a s a de Clostridium acetobutylicum en el v e c to r de e xp re s io n p B A D 24 ba jo el s is tem a p ro m o to r prpBCDE. P ara p ro d u c ir bu tano l u o tros a lcoh o le s de cadena corta , o tros o rg an ism os h o sp e d a d o re s de p ro du cc ion pu ed en m o d ifica rse de m an e ra s im ila r. P or e jem p lo , en un h o s p e d a d o r de p roducc ion pued e p ro du c irse e ta no l u tiliza n d o los m e to d o s de sc rito s po r K a lsch e u e r e t al., M ic ro b io lo gy , 152 :2529 -2536 , 2006.

In addition to going to Lcoh or following them by side A, the older person can go to the hospital or others to him or her from Cad in a co rta ta le s co moe tano l, p clothingno l, iso p ro pan ol, Isob u tano ly bu tano l for your in co rp o ra tio n on the side A u use the tea cn ic as well as id So in the morning. P o jem p lo, the o rg an ism oho sp age of p ro du cc io n can p ro duc go bu tano l. P ara c re ar ce lu las p ro duc to ra s de bu tano l, strain LS 9001, po re jem p lo, can genoman ip u la rse ad ic io na lm in te para ra e xp re sa r atoB (a ce til-C or A a ce tiltra ns fe ra sa) from Escherichia coli K12, p-h id ro x ib u tir il-C or A from sh id ro gena sa from Butyrivibrio fibrisolvens, c ro to na sa of Clostridium beijerinckii, bu tir il C o A of sh id ro gena sa of Clostridium beijerinckii, to ld eh id ode sh id ro gena sa (ALDH) of ac ilac ion of C or A of Cladosporium fulvum and adhE which co d ifica one to ld eh id or lco holdesh id ro genasa from Clostridium acetobutylicum in the neighborhood of xp re s io np BAD 24 under the s is tem ap ro mo to r prpBCDE. P o p ro duc go bu tano luo tros a lcoh o le s of short chain, or tros o rg an ism os ho sp edado re s de p ro du cc ion can be modified from m an e ra s im ila r. P o e jem p lo, in a host of production, it can be used not to use the methods of the rite s K r K a lsch eueret al., M ic ro b io lo gy, 152: 2529-2536, 2006.

III. Ingenieria genetica de cepas de produccion para aumentar/mejorar la produccion/rendimiento de derivados de acidos grasos

III. Genetic engineering of production strains to increase / improve the production / performance of fatty acid derivatives

En una celula hospedadora de produccion, utilizando tecnicas conocidas en la materia, pueden introducirse de forma estable o transitoria secuencias de polinucleotidos heterologas implicadas en una ruta biosintetica para la produccion de derivados de acidos grasos. Como ejemplos no limitativos de dichas tecnicas se incluyen electroporacion, precipitacion con fosfato de calcio, transfeccion mediada con DEAE-dextrano, transfeccion mediada por liposomas, conjugacion, transduccion e integracion genomica. Para la transformacion estable, una secuencia de a Dn puede incluir ademas un marcador de seleccion, incluyendo, por ejemplo, marcadores de resistencia a antibioticos, y genes que complementan deficiencias auxotroficas. Por otro lado, los polinucleotidos endogenos implicados en la ruta biosintetica para la produccion de derivados de acidos grasos tambien pueden mutarse utilizando tecnicas de alteracion genomica conocidas. Estas estrategias pueden aplicarse individualmente o en combinacion.

In a production host cell, using techniques known in the art, heterologous polynucleotide sequences involved in a biosynthetic pathway for the production of fatty acid derivatives can be introduced stably or transiently. Non-limiting examples of such techniques include electroporation, precipitation with calcium phosphate, transfection mediated with DEAE-dextran, transfection mediated by liposomes, conjugation, transduction and genomic integration. For stable transformation, an a Dn sequence may further include a selection marker, including, for example, antibiotic resistance markers, and genes that complement auxotrophic deficiencies. On the other hand, the endogenous polynucleotides involved in the biosynthetic pathway for the production of fatty acid derivatives can also be mutated using known genomic alteration techniques. These strategies can be applied individually or in combination.

Diversos aspectos del presente documento utilizan un vector de expresion que incluye una secuencia de ADN heterologa que codifica una proteina implicada en una ruta metabolica o biosintetica. Los vectores de expresion adecuados incluyen, pero sin limitacion, vectores viricos (tales como vectores de baculovirus), vectores de fagos (tales como vectores de bacteriofagos), plasmidos, fagemidos, cosmidos, fosmidos, cromosomas bacterianos artificiales, vectores viricos (por ejemplo, vectores viricos basados en virus variolovacunal, poliovirus, adenovirus, virus adenoasociados, SV40, virus del herpes simple, y similares), cromosomas artificiales basados en P1, plasmidos de levadura, cromosomas artificiales de levadura, y cualquier otro vector para hospedadores de produccion especificos de interes (tales como E. coli, Pseudomonas pisum y Saccharomyces cerevisiae).

Various aspects of the present document use an expression vector that includes a heterologous DNA sequence that encodes a protein involved in a metabolic or biosynthetic pathway. Suitable expression vectors include, but are not limited to, viral vectors (such as baculovirus vectors), phage vectors (such as bacteriophage vectors), plasmids, phagemids, cosmids, phosphides, artificial bacterial chromosomes, viral vectors (e.g., viral vectors based on variolovacunal virus, poliovirus, adenovirus, adeno-associated virus, SV40, herpes simplex virus, and the like), artificial chromosomes based on P1, yeast plasmids, artificial yeast chromosomes, and any other vector for specific production hosts of interest (such as E. coli, Pseudomonas pisum and Saccharomyces cerevisiae).

Los vectores de expresion utiles pueden incluir uno o mas genes marcadores de seleccion para proporcionar un rasgo fenotipico para la seleccion de celulas hospedadoras de produccion transformadas. Un gen marcador de seleccion codifica una proteina necesaria para la supervivencia y el crecimiento de celulas hospedadoras que crecen en un medio de cultivo selectivo. Las celulas hospedadoras de produccion no transformadas con el vector que contiene el gen marcador de seleccion no sobreviviran en el medio de cultivo. Los genes de seleccion tipicos codifican proteinas que (a) confieren resistencia a antibioticos o a otras toxinas (por ejemplo, ampicilina, neomicina, metotrexato o tetraciclina); (b) complementan deficiencias auxotroficas; o (c) suministran nutrientes criticos no disponibles en medios complejos (por ejemplo, el gen que codifica el racemato de D-alanina de Bacilli). En aspectos alternativos, el gen marcador de seleccion uno que codifica dihidrofolato reductasa o confiere resistencia a neomicina (para su uso en cultivos de celulas eucariotas), o uno que confiere resistencia a tetraciclina o ampicilina (para su uso en una celula hospedadora de produccion procariota, tal como en E. coli).

Useful expression vectors may include one or more selection marker genes to provide a phenotypic trait for the selection of transformed production host cells. A selection marker gene encodes a protein necessary for the survival and growth of host cells that grow in a selective culture medium. Production host cells not transformed with the vector containing the selection marker gene will not survive in the culture medium. Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins (eg, ampicillin, neomycin, methotrexate or tetracycline); (b) complement auxotrophic deficiencies; or (c) supply critical nutrients not available in complex media (for example, the gene encoding the Bacilli D-alanine racemate ). In alternative aspects, the selection marker gene one that encodes dihydrofolate reductase or confers resistance to neomycin (for use in eukaryotic cell cultures), or one that confers resistance to tetracycline or ampicillin (for use in a prokaryotic production host cell , as in E. coli).

En el vector de expresion, la secuencia de ADN que codifica el gen en la ruta biosintetica esta unida operativamente a una secuencia de control de expresion apropiada (por ejemplo, promotores, potenciadores, y similares) para dirigir la sintesis del producto genico codificado. Dichos promotores pueden obtenerse de fuentes microbianas o viricas, incluyendo, por ejemplo, de CMV y SV40. Dependiendo del hospedador de produccion/sistema de vector utilizado, en el vector de expresion se puede utilizar cualquier numero de elementos de control de transcripcion y traduccion adecuados, incluidos promotores constitutivos e inducibles, elementos potenciadores de la transcripcion, terminadores de transcripcion, y similares. Vease, por ejemplo, Bitter et al., Methods in Enzymology, 153:516-544, 1987.

In the expression vector, the DNA sequence encoding the gene in the biosynthetic pathway is operatively linked to an appropriate expression control sequence (eg, promoters, enhancers, and the like) to direct the synthesis of the encoded gene product. Such promoters can be obtained from microbial or viral sources, including, for example, from CMV and SV40. Depending on the production host / vector system used, any number of suitable transcription and translation control elements, including constitutive and inducible promoters, transcription enhancing elements, transcription terminators, and the like can be used in the expression vector. See, for example, Bitter et al., Methods in Enzymology, 153: 516-544, 1987.

Como promotores adecuados para su uso en celulas hospedadoras procariotas se incluyen, pero sin limitacion, promotores capaces de reconocer polimerasas de T4, T3, Sp6 y T7, los promotores Pr y Pl del bacteriofago lambda, los promotores de E. coli, trp, recA, de choque termicolacZ, los promotores de alfa-amilasa y especificos de sigma de B. subtilis, los promotores de los bacteriofagos de Bacillus, promotores de Streptomyces, el promotor int del bacteriofago lambda, el promotor bla del gen de beta-lactamasa de pBR322 y el promotor CAT del gen de cloranfenicol acetil transferasa. Los promotores procariotas se revisan en Glick, J. Indust. Microbiol., 1:277, 1987; Watson et al., MOLECULAR BIOLOGY OF THE GENE, 4a Ed. (1987), Benjamin Cummins (1987); y en Sambrook et al., MOLECULAR CLONING: A LABORATORY MANUAL, 2a Ed. Cold Spring Harbor Laboratory Press, 1989). Los ejemplos no limitantes de promotores eucariotas adecuados para su uso en un hospedador de produccion eucariota son de origen virico e incluyen el promotor del gen de metalotioneina I de raton (Hamer et al., J. Mol. Appl. Gen., 1:273, 1982); el promotor TK del virus del herpes (McKnight, Cell, 31:355, 1982); el promotor temprano de SV40 (Benoist et al., Nature, 290:304, 1981); el promotor de citomegalovirus (Foecking et al., Gene, 45:101, 1980); el promotor del gen gal4 de levadura (Johnston et al., PNAS (EE. UU.), 79:6971, 1982; Silver et al., PNAS (u Sa ), 81:5951, 1984); y el IgG promotor (Orlandi et al., PNAS (EE. UU.), 86:3833, 1989).

Suitable promoters for use in prokaryotic host cells include, but are not limited to, promoters capable of recognizing T4, T3, Sp6 and T7 polymerases, the Pr and Pl promoters of the bacteriophage lambda, the promoters of E. coli, trp, recA , of thermal shock lacZ, the alpha-amylase and sigma-specific promoters of B. subtilis, the bacteriophage promoters of Bacillus, promoters of Streptomyces, the int promoter of the bacteriophage lambda, the bla promoter of the beta-lactamase gene of pBR322 and the CAT promoter of the chloramphenicol acetyl transferase gene. Prokaryotic promoters are reviewed in Glick, J. Indust. Microbiol., 1: 277, 1987; Watson et al., MOLECULAR BIOLOGY OF THE GENE, 4th Ed. (1987), Benjamin Cummins (1987); and in Sambrook et al., MOLECULAR CLONING: A LABORATORY MANUAL, 2nd Ed. Cold Spring Harbor Laboratory Press, 1989). Non-limiting examples of eukaryotic promoters suitable for use in a eukaryotic production host are of viral origin and include the promoter of the mouse metallothionein I gene (Hamer et al., J. Mol. Appl. Gen., 1: 273 , 1982); the herpes virus TK promoter (McKnight, Cell, 31: 355, 1982); the SV40 early promoter (Benoist et al., Nature, 290: 304, 1981); the cytomegalovirus promoter (Foecking et al., Gene, 45: 101, 1980); the yeast gal4 gene promoter (Johnston et al., PNAS (USA), 79: 6971, 1982; Silver et al., PNAS (u Sa), 81: 5951, 1984); and the IgG promoter (Orlandi et al., PNAS (USA), 86: 3833, 1989).

El hospedador de produccion puede genomanipularse (modificarse mediante ingenieria genetica) con una secuencia genica heterologa que codifica un producto genico de la ruta biosintetica que esta unido operativamente a un promotor inducible. Los promotores inducibles son conocidos en la tecnica. Como ejemplos no limitativos de promotores inducibles adecuados se incluyen promotores que son afectados por proteinas, metabolitos, o productos quimicos. Estos incluyen, pero sin limitacion: un promotor del virus de la leucemia bovina, un promotor de la metalotioneina, un promotor de MMTV inducible por dexametasona, un promotor de SV40, un promotor polIII de MRP, un promotor de CMV inducible por tetraciclina (tal como el promotor de CMV inmediato temprano humano) asi como los operones trp y lac.

The production host can be genomanipulated (modified by genetic engineering) with a heterologous gene sequence encoding a gene product of the biosynthetic pathway that is operably linked to an inducible promoter. Inducible promoters are known in the art. Non-limiting examples of suitable inducible promoters include promoters that are affected by proteins, metabolites, or chemicals. These include, but are not limited to: a bovine leukemia virus promoter, a metallothionein promoter, a dexamethasone inducible MMTV promoter, an SV40 promoter, an MRP polIII promoter, a tetracycline inducible CMV promoter (such as the human CMV immediate early promoter) as well as the trp and lac operons .

En a lgu nos e jem p los , un h o sp e d a d o r de p ro du cc ion se g e n o m a n ip u la con una secu en c ia gen ica h e te ro loga que co d ifica un p ro du c to gen ico de la ruta b ios in te tica que esta un ido o p e ra tiva m e n te a un p ro m o to r con s titu tivo . Los p ro m o to re s con s titu tivo s a d ecu ad os son con oc idos en la te c n ic a e inc luyen el p ro m o to r ta rd io p rinc ip a l de a d e n o v iru s con s titu tivo , un p ro m o to r co n s titu tivo de M P S V , o un p ro m o to r co n s titu tivo de C M V .

In some years, a ho age of p r du duction is gen ipad with a se se in the genetics which has been coded as a genetics p Basic route that is an operational state at a p ro mo t r with the substitute. Those with sad sadness are with their eyes on the tea cn ic ae inc they look for the right way to adenov iru s with the substitute, a For the MPSV constituent, or a CMV constituent.

En a lg u n o s e jem p los, un h o s p e d a d o r de p ro du cc ion m o d ifica d o es uno que es ta g e n o m a n ip u la d o con una secu en c ia g e n ica exogena que c o d ifica una so la p ro te ina im p lica da en una ru ta de b ios in tes is . En o tros aspec tos , un h o s p e d a d o r de p ro du cc ion m o d ifica d o es uno que es ta g e n o m a n ip u la d o con s e cu e n c ia s de genes e xo ge no s que cod ifica n dos o m as p ro te in a s im p lica d a s en una ruta de b ios in tes is , po r e jem p lo , la p rim e ra y seg un da e n z im a s en una ru ta b ios in te tica .

In a few years, a modified host is one that is genome ip u do with an exogenous gene sequence that encodes a single object. Implication is given in a b ios tes route. In other respects, a modified host is one that is gen gen ip with the gene genes that encodes two or more other genes. In the same way, in a binary route, the first one, and the first day of an enzyme in a technical route.

C u a n d o un h o sp e d a d o r de p ro du cc ion se g e n o m a n ip u la para e xp re sa r dos o m as p ro te inas im p lica d a s en una ruta b ios in te tica , es tas se cu e n c ia s ge n icas pueden e s ta r co n te n id a s en un so lo v e c to r de exp res ion o en ve c to re s de e xp res ion d is tin tos . C u an do esas s e cu e n c ia s ge n icas estan co n te n id a s en un so lo v e c to r de exp res ion , en a lgu nos asp ec tos , las se cu e n c ia s de p o lin u c le o tid o s esta ran un idas o p e ra tiva m e n te a un e le m e n to de con tro l com un en do nde d icho e lem e n to con tro la la exp res ion de to d a s las se cu e n c ia s ge n icas que cod ifican las p ro te inas de la ruta b io s in te tica en el un ico v e c to r de exp res ion (p o r e jem p lo , un p rom otor).

When an age of first generation is generated, it is generated for two or more years in a basic route, it is important to know Genres can be both in only one of the neighboring regions or in view of the different regions. When you know what genres are common in only one of the neighboring regions, in some cases you will find out about them. They are going to be an operative step to a common state where they say where they are with the expiration of all the sentences. genes encoding the pathways of the basic in-route route in the only neighborhood of expiration (for example, a grantor).

C u a n d o un h o s p e d a d o r de p ro du cc ion m o d ifica d o se g e n o m a n ip u la con s e cu e n c ia s de A D N he te ro lo g a s que cod ifica n dos o m as p ro te inas im p lica da s en una ruta de b ios in tes is , una de las se cu e n c ia s de A D N puede e s ta r un id a o p e ra tiva m e n te a un p ro m o to r induc ib le , y una o m as de las s e cu e n c ia s de A D N pueden e s ta r un idas o p e ra tiva m e n te a un p ro m o to r con s titu tivo .

When a modified host host is generated by the DNA group, the gas that encodes two or more other implicit lines on a route In other words, one of the DNA sequences may be a positive operative ID for a inductive purpose, and one of the more important ones. DNA may be an operative step to a substitute with a substitute.

En a lgu nos aspec tos , la co n ce n tra c io n in tra ce lu la r (es decir, la co n cen tra c ion en el h o sp e d a d o r de p roducc ion g e n o m a n ip u la d o ) de un p ro du c to in te rm ed io de una ruta b ios in te tica , puede a u m e n ta rse para re fo rza r m as el re n d im ie n to de l p ro du c to fina l. La c o n ce n tra c io n in tra ce lu la r de l p ro du c to in te rm ed io puede a u m e n ta rse de va ria s m aneras , in c luye nd o , pe ro sin lim itac ion , el a u m e n to de la co n ce n tra c io n en el m ed io de cu ltivo de un sus tra to para una ruta b ios in te tica ; el a u m e n to de la ac tiv ida d ca ta litica de una en z im a que es a c tiva en la ru ta b ios in te tica ; el a u m e n to de la ca n tid a d in tra c e lu la r de un su s tra to (p o r e je m p lo ,u n su s tra to p r im a rio ) pa ra una e n z im a qu e es a c tiva en la ru ta b ios in te tica ; y s im ila res.

In some respects, the intra c tion of the river (that is, the center of the genoman ip u do r) of a gen In order to carry out a basic route, it can be increased in order to reinforce the imitation of the end of the year. The con- ce ntra tio n in t e ce r r l lp ro du c to in te rm ed io can be increased in several ways, in c luye nd o, but without limitation, the increase The co-ce ntra tio n in the quantitative environment of a deal for a basic technical route; the year of the critical activity of one en z im a that is active in the technical route; The increase in the amount of money in a state (for example, a first step) for an enzyme that is active in the b a bios in te tica; and s im ila res.

En a lg u n o s e jem p los , el d e riva d o de ac ido g ra so o p ro d u c to in te rm e d io se p ro d u ce en e l c ito p la sm a de l h o s p e d a d o r de p ro du cc ion . La co n ce n tra c io n c ito p la sm a tica se puede a u m e n ta r de va ria s m aneras, in c luye nd o , pero sin lim itac ion , union del ac ido g raso a la coe nz im a A para fo rm a r un a c il-C o A tioe s te r. A d ic io n a lm e n te , la con cen tra c ion de a c il-C o A se puede a u m e n ta r al a u m e n ta r la b io s in te s is de C o A en la ce lu la , ta l co m o m e d ia n te la so b re e xp re s io n de ge ne s a s o c ia d o s con la b io s in te s is de p a n to te n a to (p o r e jem p lo , panD) o in a c tiva c io n de ge ne s a so c ia d o s con la b ios in tes is de g lu ta tio n (p o r e jem p lo , g lu ta tio n s in tasa).

In some years, the riva do de ac gone g ra so op ro duc to in te rm ed io du p ce du ce in elc ito p la sm a of the p ro du cc ion host. The relationship with the symphony can be increased in several ways, but without limitation, union of the mainland to the coefficient of the coefficient. A for fo rm ar an ac il-C or A tioe s te r. On a daily basis, the ac-C or A con - traction can be increased by increasing the number of C or A in the city, The co-mother is the only one of the two members of the country with the bureau only of bread (for example, pan , d) or in ac tiva c In general, with the b ios ies tes de g lu ta tio n (pore jem p lo, g lu ta un tio ns in rate).

En el c ro m o so m a de l h o sp e d a d o r de p ro du cc ion pu ed en in tro d u c irse o a lte ra rse s e cu e n c ia s regu lado ras , se cu e n c ia s cod ifica n tes , y c o m b in a c io n e s de las m ism as. En a lgu nos e jem p los , la in te g ra c ion de la secu en c ia reco m b in an te de sea da en la se cu e n c ia ge n o m ica del h o sp e d a d o r de p roducc ion no requ ie re el uso de un m a rca d o r de se lecc ion ta l com o un an tib io tico . En a lgunos e jem p los , las a lte rac ione s g e n o m ica s in c luyen el cam b io de la secu en c ia de con tro l de los genes d iana al re e m p la za r el p ro m o to r (o los p ro m o to re s ) na tivo con un p ro m o to r que es in sen s ib le a la regu lac ion . E x is ten n u m e ro sa s e s tra te g ia s para h a ce r esto . P o r e jem p lo , V a lle y F lo res, en M e tho ds Mol. Biol., 267 :113 -122 , 2006 , de sc rib en un m e tod o basado en P C R para so b re e x p re s a r genes c ro m o so m ico s en E. coli. O tra e s tra te g ia se basa en el uso de o lig o n u c le o tid o s m o n o ca te n a rio s pa ra c re a r m u ta c io n e s e sp e c ifica s d ire c ta m e n te en el c rom o som a , u tiliza n d o la te c n ic a d e sa rro lla d a po r C o u rt e t al., P N A S (U S A ), 100 :15748 -15753 , 2003. E sta te cn ica se basa en el uso de la so b re e x p re s io n de la p ro te ina B eta de l b a c te rio fa g o la m bd a pa ra p o te n c ia r la re co m b in a c io n gene tica . C o m o v e n ta ja s de es ta e s tra te g ia se inc luyen que pu eden u tiliza rse o lig o n u c le o tid o s s in te ticos de 70 bases de lo ng itud (o m as) para c re a r m u tac ion es , in se rc io n e s y d e le c ion es pun tua les , e lim in a n d o a s i cu a lq u ie r e ta pa de c lo na c ion . A s im ism o , el s is te m a es lo s u fic ie n te m e n te e ficaz com o para que no sean n e cesa rios m a rca d o re s para a is la r las m u ta c io n e s deseadas .

In the case of age, the age of the population can be left in advance or when it is regulated, it is codified, and it is agreed upon. io nes of the same. In some cases, the integration of the sectoral sector into the mainstream in the mainstream of the old age of non-production. It requires the use of a common reading instrument or a tibiotic sensor. In some of them, the most important racial groups in the sector change with each other of the genes from the day to the start of the project. to r (or the p ro mo to re s) with a p ro mo to r which is in sen s ib le to the region. This is the number of people to do this. P ore jem p lo, V a lle and F lo res, in M e tho ds Mol. Biol., 267: 113-122, 2006, of sc ribs in a PCR-based method for undergoing only genes such as E. coli. Another step is based on the use of the link to the national government for the armed forces of the civil society in the Roman Republic. The use of the tea cn ic ade sa rro lla da po r C ou rt et al., PNAS (USA), 100: 15748-15753, 2003. This technique is based on the use of the so b re exp re s io n de la p ro te ina B eta de lbac laughed fa mbd a pa ra po te nc ia r the re co mb in gene ation. How many of these are included is that they can be used or linked to the 70 bases of the technical bases (or more) for the economic structure In se rc io nesyde le c ion es pun tuales, and lim in andoas i cu a lq u ie re ta pa de l lo nation. In addition, the system is enough to be effective so that they are not necessary to make the desired mutation.

C on esta e s tra te g ia , la reg ion reg u la d o ra de un gen pued e ca m b ia rse pa ra c re a r un p ro m o to r m as fu e rte y /o e lim in a r el s itio de un ion de un rep reso r. P o r con s ig u ie n te , en el o rg a n ism o h o sp e d a d o r de p ro du cc ion puede s o b re e xp re sa rse un gen deseado .

With this in mind, the regional region of a gene may be able to go a step further and / or be limited to the site. of an ion of a rep r. P o r s s s u u n n e, in o rg a n ism o h o sp e d a d o r p p du du tion can be a desired gene.

IV. Fermentacion

IV. Fermentation

A. Maximizacion de la eficacia de produccion

A. Maximization of production efficiency

La p ro d u cc io n y el a is la m ie n to de d e riv a d o s de ac ido s g ra sos pu eden m e jo ra rse e m p le a n d o te c n ic a s e sp e c ifica s de fe rm e n ta c io n . Un m etod o para m a x im iz a r la p ro du cc ion al m ism o tie m p o que reduce costes, es a u m e n ta r el p o rce n ta je de la fu e n te de ca rb o n o que se tra n s fo rm a en p ro du c to s de h id roca rbu ro .

The law of the day and the year is the way of life of the people that can be improved by the technology of faith in the faith in the world. . A method to maximize the cost at the same time as to reduce costs is to increase the power of the source of money that is being processed. a p ro du c to s of h id roca rbu ro.

D u ran te los c ic los de v id a ce lu la res no rm a les , el ca rb o n o se u tiliza en las fu n c io n e s ce lu la re s para p ro d u c ir lip idos, saca rid os , p ro te inas, ac ido s o rg a n ico s y p o lin u c le o tid o s . La red ucc ion de la can tidad de ca rb on o ne cesa ria para las a c tiv id a d e s re la c io n a d a s con el c rec im ien to , pu ede a u m e n ta r la e fica c ia de la tra n s fo rm a c io n de la fu e n te de ca rbono a la p ro du cc ion . E sto pu ede rea liza rse cu ltiva n d o p rim e ro los m ic ro o rg a n is m o s a una de ns idad deseada , lo cua l se rea liza en el p ico de la fase lo ga ritm ica de cu ltivo . D espues, para d e te n e r el c re c im ie n to de las ce lu las , pueden a p ro ve ch a rse genes del pun to de con tro l de rep licac ion . E spe c ificam en te , para a c tiva r genes aso c ia dos a la fase es ta c ion a ria , pueden u tiliza rse m e ca n ism o s de pe rcep c ion de cu o ru m (quorum sensing mechanisms), (segun lo rev isad o en C am illi y B ass le r, S c ience , 311 :1113 , 2006; V en tu ri, F E M S M icrob io . R ev., 30 :274 -291 ,2006 ; y R ead ing y S pe ran d io , B M C M icrob io l. Lett., 254: 1-11,2006).

In the cases of the residents of the region, the channel is used in the census operations for the purpose of lipids, it will be ridiculous. But, here, I am only so friendly. The network of the amount of money needed for the current activities with the newcomer can increase the efficiency of the work. rm ac io n of the f in t he carbon in the p ro du cc ion. This can be done quantitatively in the same way as the desired one, which is done at the peak of the rhythmic phase of quantitative phase. Then, in order to have the reference of the cells, genes from the replication point can be used. Specifically, in order to activate genes associated with the phase, they can also be used in terms of how to quorum ( quorum sensing mechanisms). , (as reviewed in C am illi and B ass le r, S c ience, 311: 1113, 2006; V in your ri, FEMSM icrob io. R ev., 30: 274-291, 2006; and R ead ing and S pe ran d io, BMCM icrob io L. Lett., 254: 1-11,2006).

G enes que pueden a c tiva rse para d e te n e r la re p lica c io n y el c re c im ie n to c e lu la r en E. coli in c luye n genes umuDC, cuya so b re e xp re s io n d e tie n e la p ro g re s ion de la fase e s ta c io n a ria al c re c im ie n to e x p o n e n c ia l (M u rli e t al., J. o f B act., 182 :1127 , 2000 ). U m u C es una A D N p o lim e ra sa que pu ede lle va r a cab o la s in te s is de tra n s le s io n sob re le s ion es no co d ifica n te s - la base m e ca n ica de la m a yo ria de las m u ta g e n e s is U V y qu im ica . Los p ro du c to s ge n icos umuDC se u tilizan para el p ro ceso de s in tes is de tra n s le s io n y ta m b ie n s irven com o pun tos de con tro l de da no s en secu en c ias de po lin u c le o tid o s . Los p ro du c to s ge ne ticos umuDC in c luyen U m uC , U m uD , um uD ', U m u D '² C, U m u D '² y /o U m u D ². M ie n tras tan to , los genes p ro du c to res de p ro du c to pueden ac tiva rse , m in im iza nd o as i la ne ces idad de u tiliza r ru tas de rep lica c io n y m a n te n im ie n to m ie n tras se p ro du ce el de riva d o de ac ido graso . Los m ic ro o rg a n ism o s h o sp e d a d o re s de p ro du cc ion ta m b ie n pueden g e n o m a n ip u la rse para e xp re sa r umuC y /o umuD de E. coli en p B A D 24 ba jo el s is te m a p ro m o to r prpBCDE a tra v e s de la s in te s is de novo de es to s ge ne s con los ge ne s de p ro du cc ion de l p ro du c to fina l ap rop iad os .

Genes that can be activated in order to have the p ricality and the c re c i n e ce to the r in E. coli in c luye n umuDC genes , whose so b re e xp re s io Where the p ro g reion of the phase is also the exposition (M u rli et al., J. of B act., 182: 1127, 2000 ). U mu C is a small DNA that can be carried out, or the only thing that is left behind is not co-ifica ned - the base of the The only way to do it is UV and chemical chemistry. The umuDC genetics are used for the period of syndromes that are of interest and are also dealt with as damages in the area. in po ias de po lin uc le o tid os. The genetic factors umuDC in c include U m uC, U m uD, um uD ', U mu D' ² C, U mu D ' ² and / or U mu D ² . In the meantime, the pro ductive genes can be activated, minimized, and thus the need for use of public relations policies. I n im ie n to me after the riva do of fatty acid. The m iro or rg an ism osho sp edado re s de p ro du cc ion ta mb ie n can genoman ip u la rse for e xp re sa r umuC and / or umuD of E. coli in p BAD 24 ba jo the s is te map ro mo to r prpBCDE through de s sin te s is de novo de es to s ge ne s with the ge ne s of p ro du cc ion of lp ro du c to fina l ap rop iad os.

El po rcen ta je de c a rb o n e s de e n tra da tra n s fo rm a d o s en e s te re s g ra sos o en p ro du c to s de h id ro ca rbu ro es un in d u c to r de costes. C u an to m as e ficaz sea el p ro ceso (es decir, cua n to m a yo r sea el po rcen ta je de ca rb o n e s de e n tra da tra n s fo rm a d o s en es te res g ra sos o p ro du c to s de h id ro ca rbu ro , m en os cos toso sera el p roceso . P ara las fu e n te s de ca rb o n o que con tien en ox ig e n o (p o r e jem p lo , g lu co sa y o tras fu e n te s ba sad as en h id ra to s de ca rbono), el o x ig eno se libe ra en fo rm a de d iox id o de ca rbono . P o r cada 2 a tom os de o x ig eno libe rados , ta m b ie n se libe ra un a to m o de ca rb on o , lo que con du ce a una e fica c ia m e ta b o lica te o ric a m ax im a de a p ro x im a d a m e n te 34 % (p /p ) (pa ra p ro du c to s d e riva dos de ac ido s grasos). S in em bargo , esta cifra, cam b ia para o tros p ro du c to s de h id ro ca rbu ro y fu e n te s de ca rbono . Las e fica c ia s tip ica s en la b ib liog ra fia son a p ro x im a d a m e n te <5 %. Los ho sp e d a d o re s de p ro du cc ion g e n o m a n ip u la d o s para p ro d u c ir p ro du c to s de h id ro ca rbu ros , pueden te n e r una e fica c ia s u p e rio r a a p ro x im a d a m e n te 1 %, p o r e jem p lo , una e fica c ia m a y o r que a p ro x im a d a m e n te 3 %, 5 %, 10 %, 15 %, 20 %, 25 % o 30 % . En un e jem p lo , los h o sp e d a d o re s de p ro du cc ion m os tra ra n una e fica c ia de a p ro x im a d a m e n te 10 % a a p ro x im a d a m e n te 25 %. En o tros e jem p los, d icho s h o spe da do re s de p ro du cc ion m os tra ra n una e fica c ia de a p ro x im a d a m e n te 25 % a a p ro x im a d a m e n te 30 %. En o tros e jem p los , d ichos ho sp e d a d o re s de p roducc ion m os tra ra n una e fica c ia > 30 %.

The power of the bank to be carried out in these days or in the case of human rights is a cost indent. How much more effective is the process (that is, how much I can be in charge of the work of those who are in this situation?) This is the process that will be the p rocess for those who have been in charge of the situation (for example, for example). After I am sad, I am sad, in the id of all carbon), the oxygen is released in the form of a carbon dioxide. ig eno freed, also a year of distribution is released, which leads to an effective physical or economic experience 34% (p / p) (for two years of fatty acids), however, this figure changes for others in the ID ro ca rbu ro y fu en te s de ca rbono The typical ethics in the bio liog ra fia are ap ro x im adamen te <5%. cc gene ion oman ip u the two for p ro duc go p ro du c to s of h id ro ca rbu ros, they can have an e ra tica r io rio raap ro x im adamen te 1%, pore jem p lo, an e fica c ia greater than ap ro x im adamen te 3%, 5%, 10%, 15%, 20%, 25% or 30%. In one example, the number of years of return will be an effective 10% to 10% a year to 25%. In other cases, that is only a few years ago, it will have an effective 25% a year of 30%. In other cases, these cases are more than 30% effective.

El h o s p e d a d o r de p ro du cc ion puede g e n o m a n ip u la rse ad ic io n a lm e n te para e xp re sa r ce lu lo so m a s reco m b in an te s , com o los desc ritos en la so lic itud P C T nu m ero P C T /U S 2007 /003736 , lo que puede p e rm itir al p ro d u c to r de p ro du cc ion u tiliza r m a te ria l ce lu lo s ico com o fue n te de ca rb on o . P o r e jem p lo , el h o s p e d a d o r de p ro du cc ion puede g e n o m a n ip u la rse a d ic io n a lm e n te para e x p re s a r in ve rta sa s (E C 3.2.1.26 ) pa ra que la s a ca ro sa pued a u tiliza rse com o fu e n te de carbono .

The host of the pipeline can gen ip ip the ad ad io io na lm en te for e xp re sa r ce lu so most reco mb in an te s, as the decrees in the PCT so PCT / US 2007/003736 number, which may be allowed to be used as a general purpose. P o jem p lo, the host of p ro du cc ion can genoman ip u la ad ad io na lm en te to exp re sar in ve rta sa s (EC 3.2.1.26) so that the sa ca ro sa It can be used as carbon in carbon.

De m a n e ra s im ila r, el h o sp e d a d o r de p ro d u cc io n pued e g e n o m a n ip u la rs e u tiliza n d o las e n se n a n za s d e sc rita s en las p a te n te s de EE. UU. Nos. 5 000 000; 5 028 539; 5 424 202; 5 482 846; y 5 602 030, de m od o que el h o s p e d a d o r de p ro du cc ion pu eda a s im ila r el ca rb o n o de m an e ra e fica z y u tiliza r m a te ria le s c e lu lo s ico s com o fu e n te s de carbono . En un e jem p lo , la cam a ra de fe rm e n ta c io n con tien e un c ic lo /m e zc la de fe rm e n ta c io n que esta e x p e rim e n ta n d o una red ucc ion con tinua . En este caso , se crea un e n to rno re d u c to r es tab le . El eq u ilib rio de e le c tro n e s se m an tien e g ra c ia s a la libe ra c io n de d iox id o de ca rb on o (en fo rm a gaseosa). Los e s fu e rzos para a u m e n ta r el equ ilib rio en tre N A D /H y N A D P /H ta m b ie n pu ed en fa c ilita r la e s ta b iliza c io n de l eq u ilib rio e lec tron ico .

From m a n e ra s im ila r, the h o sp e d a d o r of p ro d u cc io can be used in the p rs a n s s s e e r rita s in the US. UU. Nos. 5,000,000; 5 028 539; 5 424 202; 5 482 846; and 5 602 030, as long as the host of the bank can thus imitate the capital of zyu tiliza rma te ria te sce lu l s sico s com o fu In carbon s. In one example, the chamber of faith rm in ta ta io n with a c iclo / me zc that of faith rm in ta that is also a network with tinua In this case, an e n to rno re d u c to r es tab le is created. The eq uterium of the le c tro n e s is also subject to the release of the tax ID or of gas (in gaseous form). The efforts are to increase the balance between NAD / H and NADP / H as well as in the Spanish state of the Spanish economy. ico.

La d ispo n ib ilida d de N A D P H in tra ce lu la r ta m b ie n puede m e jo ra rse g e n o m a n ip u la n d o el h o sp e d a d o r de p roducc ion para e xp re sa r una N A D H :N A D P H tra n sh id ro g e n a sa . La exp res ion de una o m as N A D H :N A D P H tra n sh id ro g e n a sa s tra n s fo rm a el N A D H p ro du c ido en la g luco lis is en N A D P H , lo que m e jo ra la p roducc ion de de riva dos de ac idos grasos.

The nAD ilida d of NADPH in trans ce la la r ta mb ie can be improved genoman ip u la ndo el ho sp age of p roduction for e xp re sa r a NADH: NADPH tra n sh id ro gena sa. The expiration of one or more NADH: NADPH trans a sh id ro gena sa s tra ns fo rm a la NADH p r du c gone in the g lol l lis l es in NADPH, which improves the production of of riva two fatty acids.

B. Produccion de hidrocarburos a pequena escala

B. Production of small-scale hydrocarbons

P ara la p ro du cc ion de p ro du c to s de h id ro ca rb u ro s a pe quena esca la , ce lu la s BL21 (D E 3 ) de E. coli que llevan p B A D 24 (con res is te nc ia a am p ic ilin a y la ruta de s in tes is del p ro du c to fina l), as i com o pU M V C 1 (con res is te nc ia a ka n a m ic in a y el s is tem a de s o b re e x p re s io n de a ce til C o A /m a lo n il C o A ) se in cub an d u ra n te la no che en m a tra ce s de 2 litro s a 37 °C, a g ita n d o a> 200 rpm en 500 m l de m ed io LB c o m p le m e n ta d o con 75 pg /m l de a m p ic ilin a y 50 pg /m l de kan am ic ina hasta que los cu ltivos a lcancen una D O ⁶⁰⁰ de> 0,8. D e spues de a lc a n z a r una D O ⁶⁰⁰ de> 0,8, las ce lu la s se co m p le m e n ta n con p ro p iona to de sod io 25 mM (a pH 8 ,0 ) para a c tiv a r los s is te m a s ge n icos g e n o m a n ip u la d o s para la p ro du cc ion y para d e te n e r la p ro life ra c io n c e lu la r m ed ia n te la a c tiva c io n de las p ro te inas U m u C y U m uD . La e ta pa de indu cc ion se rea liza du ran te 6 ho ras a 30 °C. D e spu es de la incubac ion , el m ed io se exa m in a pa ra d e te rm in a r el p ro du c to de h id ro ca rb u ro u tilizan do G C -M S .

P o the p ro du cc ion of p ro du c to s of h id ro ca rb u ro sa pe quena esca la, c e lu s s BL21 (DE 3) of E. coli carrying p BAD 24 (with res It is necessary for am p ic ilin a and the route of syntheses is of the p du du c to fina l), as well as p u MVC 1 (with res is te nc ia a ka nam ic in ay el s is The subject of surplus expiration of a C or C / A / m or C or A) is not covered in the case of 2 liters per 37 ° C, Aggregate at> 200 rpm in 500 ml of the LB compliant with 75 pg / ml of amp and light and 50 pg / ml of kan am ic ina until the quantitative ones reach an OD ⁶⁰⁰ of > 0.8. After an OD ⁶⁰⁰ of> 0.8 has been established, the decisions are made with 25 mM sodium pressure (at pH 8.0) for ac tiv ar The most common genoman ip u the two for the p r du tion and for the continuation of the p r life is the lu rm ed ia n the active t of the p ro you leave U mu C and U m uD. The induction stage is carried out for 6 hours at 30 ° C. After the incubation, the environment will be examined for the purpose of the use of GC-MS.

C. Produccion de hidrocarburos a gran escala

C. Large-scale hydrocarbon production

P ara la p ro du cc ion de p ro du c to s a g ran esca la , los ho sp e d a d o re s de p ro du cc ion g e n o m a n ip u la d o s se cu ltivan en lo tes de 10 litros, 100 litros, o m as g randes ; se fe rm en tan ; y se inducen para e xp re sa r los p ro du c to s de sea do s ba sa n d o se en los ge ne s e sp e c ifico s c o d ifica d o s en los p lasm id os a p ro p ia d o s en los m ism os.

For the protection of the law, the hoped of the genoman ip are both quoted in 10 liters, 100 liters, om as g randes; it rm faith in so; and they are induced for the sake of the interests of the people who are qualified in the two countries in the two countries in the world. you.

P o r e jem p lo , ce lu las BL21 (D E 3 ) de E. coli que llevan pB A D 24 (con res is te nc ia a am p ic ilin a y la ruta de s in tes is del p ro du c to fina l), a s i com o pU M VC 1 (con res is te nc ia a kan am ic ina y el s is tem a de so b re e xp re s io n de ace til C o A /m a lo n il C oA ), se in cub an a pa rtir de un cu ltivo de sem illa de 500 m l du ran te un c ic lo de fe rm e n ta c io n de 10 litros (o un cu ltivo de sem illa de 5 litros para una fe rm e n ta c io n de 100 litros) en un m ed io LB (s in g lice ro l) que con tien e k a n a m ic in a 50 pg /m l y a m p ic ilin a 75 pg /m l a 37 °C, que se ag ita a> 200 rpm hasta que el cu ltivo a lcanza una D O ⁶⁰⁰ de> 0,8, un p roceso que no rm a lm e n te du ra a p ro x im a d a m e n te 16 horas. El m ed io de fe rm e n ta c io n se co m p le m e n ta de m an e ra c o n tin u a d a para m a n te n e r una c o n ce n tra c io n de fo s fa to de sod io de 25 m M , a pH 8,0, para a c tiv a r los s is te m a s g e n ico s g e n o m a n ip u la d o s para la p ro d u cc io n y para d e te n e r la p ro life ra c io n c e lu la r m e d ia n te la a c tiva c io n de las p ro te inas U m uC y U m uD . El m ed io ta m b ie n se c o m p le m e n ta de m a n e ra c o n tin u a d a con g lucosa para m a n te n e r una c o n ce n tra c io n de 25 g /100 ml.

P o jem p lo, ce lu the BL21 (DE 3) of E. coli that carry pB AD 24 (with respect to am p ic ilin a and the route of syntheses is of the p ro du c to fina l), as well as pU M VC 1 (with res is te nc ia a kan am ic ina and the system of so b re e xp re s io n acetil C or A / ma lo n il C oA), a 500 ml semi-quantitative quantum will be included in the course of 10 liters (or a 5-liter semi-quantitative quantitative). for a faith in the 100 liter capacity) in a LB (s in g lice ro l) medium which has a range of 50 pg / m lya ic il ilin at 75 pg / m at 37 ° C, which it is agitated at> 200 rpm until the quantum reaches an OD ⁶⁰⁰ of> 0.8, a process that does not last in about 16 hours. The mediation of faith in this period is also commended with a continuance to keep one of the 25-million-million foos fate. , at pH 8.0, in order to ac- civize the sgen te s somanoman ip u the two for the purpose of life and for the purpose of life the activity of the p u te u u u u u u u u u. The mediator is also compiled in a way with a greasy stain to maintain a concentration of 25 g / 100 ml.

D e spu es de la p rim e ra hora de inducc ion , una a licu o ta de no m as del 10 % del vo lu m e n c e lu la r to ta l, se re tira cada hora y se de ja re p o s a r sin ag itac ion , lo que a su ve z pe rm ite que el p ro du c to (o p ro d u c to s ) de h id ro ca rb u ro sub an a la sup e rfic ie , e x p e rim e n ta n d o una se p a ra c io n de fase e sp on ta ne a . El com p o n e n te de h id ro ca rb u ro se recoge y la fase a cu osa vu e lve a la cam a ra de reacc ion . La cam a ra de rea cc io n fu n c io n a de m an e ra con tinu ad a . C u ando la D O ⁶⁰⁰ cae po r de ba jo de a p ro x im a d a m e n te 0,6, las ce lu la s se ree m p la za n con un nu evo lo te de cu ltivo a p a rtir de un cu ltivo de sem illa .

After the first hour of induction, one year after no more than 10% of the volume decreases, the hour is re-drawn and it is left without ag itac ion, which in turn sees that the p du du ct (op ro duc to s) of h id ro ca rb u ro goes up to the sup e rfic ie, expe rim in so much a phase phase is sp on ta ne a. The component of h id ro ca rb u ro is collected and the phase takes place to the reaction chamber. The chamber of reaction was only a few days later. When the DO ⁶⁰⁰ falls below ap ro x im adamen te 0.6, the letters will be replaced with a new quantitative report from a quantum of sem illa.

P a ra la p ro d u cc io n de e s te re s de cera , los e s te re s de ce ra se a is lan , se lavan ra p id a m e n te en H C l 1 M y re to rn an a un pH 7 a tra ve s de un lavado e xh a u s tivo con ag ua destilada .

For the purpose of waxing, waxes are added, they are washed quickly in HC l 1 M and re r r an an at pH 7 through an ausxitive wash with distilled water.

V. Procesamiento de postproduccion

V. Postproduction processing

Los de riva dos de ac ido s g rasos p ro du c ido s d u ran te la fe rm e n ta c io n pueden sep a ra rse de los m ed io s de fe rm e n ta c io n . P ara s e p a ra r d e riva d o s de ac ido s g ra sos de m ed io s acu o so s puede u tiliza rse cu a lq u ie r te cn ica con oc ida . Un p ro ceso de se p a ra c io n e je m p la r es un p roceso de se p a ra c io n de dos fa se s (b ifas ico ). E ste p roceso im p lica la fe rm e n ta c io n de los ho sp e d a d o re s de p ro du cc ion g e n o m a n ip u la d o s en con d ic io n e s su fic ie n te s para p ro d u c ir un de riva d o de ac ido graso , p e rm itien do re co g e r el de riva d o en una fase o rg an ica , y la sep a ra c ion de la fase o rg a n ica del ca ldo de fe rm e n ta c io n acuoso . Este m e tod o puede lle va rse a la p ra c tica en e n to rno s de fe rm e n ta c io n ta n to en lo tes com o en fe rm e n ta c io n con tinua .

Those of private citizens who have faith in their faith can be separated from the world of faith. For the purposes of the ac i c a tive of these activities, it is possible to use only one technique. A short period of time is a two-month process of two phases (basic). This is the implicit process of the faith in the situation of the genome of the genome ip u the two in condi tions with their fiction for the duc one of the fatty acid source, but that of the one in a single phase or region, and the phase of the unique phase of the faith rm in aqueous ta. This method can be carried out in practice in our faiths in common faiths and faith in the future.

La se p a ra c io n b ifa s ica a p ro ve ch a la re la tiva in m isc ib ilida d de los de riva dos de ac ido s g ra sos para fa c ilita r la sep a ra c io n . " In m is c ib ilid a d " se re fie re a la in ca p a c id a d re la tiva de un co m p u e s to para d iso lve rse en ag ua y se de fine y /o d e te rm in a m e d ia n te el c o e fic ie n te de rep a rto de los co m p u e s to s . Un e xp e rto ha b itu a l en la m a te ria a p re c ia ra que e lig ie n d o un ca ldo de fe rm e n ta c io n y una fase o rgan ica de m an era que el de riva do de ac ido g ra so que se p roduce te n g a un a lto v a lo r logP, el de riva d o de ac ido g ra so se s e p a ra ra en la fase o rg a n ica en el rec ip ie n te de fe rm e n ta c io n , in c luso a c o n ce n tra c io n e s bajas.

The se b a ra b e b ica s ica s a p ro ve ch a la re la tiva in m isc ib ilida d of those of riva dos de s a se gone to facilitate the sep a ration. "In m is c ib ilid ad" refers to the inca pac id ad re tiva of a cost to isolate in ag ua and be fine and / or de rm in amed ia n te The coefficient of distribution of the costs. An e xp e rto ha b itu al in the ma ap ria ap re c ia ra e l e ein a ca t of faith rm in ta t io n and a phase or rganic of m an era that the riva do So that it is possible to pay it a year, it will be the same, it will be known only in the national phase in the region of faith. rm in ta ta io n, even in low use.

Los d e riva d o s de ac ido s g ra sos p ro d u c id o s segun las co m p o s ic io n e s , los vec to re s , las ce lu las y los m e tod os de sc rito s en este docum e n to , se ran re la tiva m e n te in m isc ib le s en el ca ldo de fe rm e n ta c io n , as i com o en el c itop la sm a . P or lo tan to , el de riva d o de ac ido g raso se reco ge ra en una fase o rg an ica ya sea in tra ce lu la rm e n te y/o e x tra ce lu la rm e n te . La re co g id a de los p ro du c to s en la fase o rg a n ica d ism in u ira el im p ac to de los d e riva d o s de ac ido s g ra sos sob re la fun c ion ce lu la r, y p e rm itira al h o s p e d a d o r de p ro d u c to r p ro d u c ir una m a yo r can tidad de p roducto d u ra n te m as tiem po.

Those of riva dos de ac ido sg ra sos p ro duc os according to the co mpos ic io nes, the neighbors, re ce lu las and the methods of sc rites in this docum, ran re la tiva men te in m isc ib le s in the ca r of faith rm ta ta io n, as well as in the city of la sm a. First of all, that of private land will be collected in one phase or in the same way either in front of the river and / or in the direction of the river. The re co g id of the p ro du c to s in the phase or r n ica n ism in u will go the im p ac t of the riva two of ac c ido sg ra sos so re re the function ce ce lu la r, ype rm it will be the host of the du du to rp ro duc to go a lot of du p du du p du du m te more time.

Los a lcoh o le s grasos, los es te res g rasos, las ce ra s y los h id ro ca rb u ro s p ro du c ido s se g u n las d ivu lg a c io n e s de este do cum e n to , pe rm iten la p ro du cc ion de com p ue s to s h o m og ene os en los que al m en os a p ro x im a d a m e n te el 60 %, 70 %, 80 %, 90 %, 91 %, 92 %, 93 %, 94 % o 95 % de los a lcoh o le s g rasos, es te res g ra sos y ceras p ro du c ido s a d e cu a d a m e n te , tien en lo ng itude s de cad en a de ca rb o n o que va ria n en m en os de a p ro x im a d a m e n te 6, m en os de a p ro x im a d a m e n te 4 o m en os de a p ro x im a d a m e n te 2 ca rbonos . E stos co m p u e s to s ta m b ie n pueden p ro du c irse de m odo que te n g a n un g rado de sa tu ra c io n re la tiva m e n te un ifo rm e , po r e jem p lo , al m en os a p ro x im a d a m e n te el 60 %, 70 %, 80 %, 90 %, 91 %, 92 %, 93 %, 94 % o 95 % de los a lcoh o le s g rasos, es te res g rasos, h id ro ca rb u ro s y ceras, es tan m on o in sa tu ra d o s , d iin sa tu ra d o s o tr iin sa tu ra d o s . E stos co m p u e s to s pueden u tiliza rse d ire c ta m e n te com o p ro du c to s o c o m p o n e n te s de p roduc tos , po r e jem p lo , com o co m b u s tib le s , de te rge n tes , lub rican tes , ad itivo s para el cu id a d o pe rsona l, c o m p le m e n to s s u p le m e n to s etc. E s tos c o m p u e s to s ta m b ie n pueden u tiliza rse co m o m ate ria prim a para rea cc io nes p o s te rio res para p re p a ra r o tros p roductos, inc luyendo , po r e jem p lo , tra n se s te rif ica c io n , h id ro gen ac ion , c ra q u e o ca ta litico (m e d ia n te h id ro gen ac ion , p iro lis is , o am bas), o rea cc io nes de epox idac ion .

The fatty ones, the fatty ones, the ce s s and the h id ro ca rb u ro sp ro du c gone s are the days of this cum in to However, in the case of shom og ene in which at least 60%, 70%, 80%, 90%, 91%, 92 %, 93%, 94% or 95% of those who are living or living, this is what they need and waxes, since they are in the same category as in each case. One that will be in me in the months of April 6, in the years of April 4 in the years of the year. These costs may also leave you in a way that will deny you a rationale for the rationale for an individual rm e, po re jem p lo, at In the case of 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94% or 95% of the years, this is the case, h id ro ca rb u ro s and waxes, it is so great on your two, say on your own two days. These costs can be used to make sure that they are composed of products, as well as bussines, of the rulers, lub rican tes, ad itivo s for the cu id ado pe rsona l, comp le men to ssup le men to s etc. These compounds can also be used as a primary concern for potential customers to produce products, including money, money, etc. It is important to know how to act in relation to the gen- etic (to the extent that the gen- ation of the gen- eration, public relations, or basic terms), or reactions of epox idac ion.

Los a lcoh o le s g rasos, los es te res g rasos , las ce ra s y los h id ro ca rb u ro s p ro d u c id o s segun las co m p o s ic io n e s , los vec to re s , las ce lu la s y los m e tod os d ivu lg ad os en este d o cu m e n to con tienen n ive les ba jos de e le m e n to s no e sp e ra d o s o no desea do s , in c luye nd o , pero sin lim itac ion , m e ta les pesados. En a lgu nos aspec tos , los a lcoh o le s g rasos, los es te res g rasos, las ce ras y los h id ro ca rbu ros p ro du c ido s com o se de sc rib e en el p resen te docum en to , con tien en a d e cu a d a m e n te m en os de a p ro x im a d a m e n te 50 ppm de a rsen ico ; m en os de a p ro x im a d a m e n te 300 ppm de ca lc io ; m en os de a p ro x im a d a m e n te 200 ppm de c lo ro; m en os de a p ro x im a d a m e n te 50 ppm de coba lto ; m en os de a p ro x im a d a m e n te 50 ppm de cob re ; m en os de a p ro x im a d a m e n te 300 ppm de h ie rro ; m en os de a p ro x im a d a m e n te 2 % en peso de agua; m enos de a p ro x im a d a m e n te 50 ppm de p lom o; m en os de a p ro x im a d a m e n te 50 ppm de m an ga neso ; m en os de a p ro x im a d a m e n te 0 ,2 ppm de m ercu rio ; m en os de a p ro x im a d a m e n te 50 ppm de m o libdeno ; m en os de a p ro x im a d a m e n te 1 % en peso de n itro ge no ; m en os de a p ro x im a d a m e n te 200 ppm de po tas io ; m en os de a p ro x im a d a m e n te 300 ppm de sod io ; m en os de a p ro x im a d a m e n te 3 % en peso de azu fre ; m en os de 50 ppm de cinc; y /o m en os de 700 ppm de fosfo ro .

The lcoh or the following, the latter, the ce s and the h id ro ca rb u ro sp ro duc id according to the commpositions, the neighbors, the neighbors s and the methods of this year in which they have low levels of income do not wish to do so, do not want to do so, but without limitation Itation, it makes me heavy. In some cases, the lcoh or the next ones, the latter, the cera and the cents are shared with the sc ribs in the past I docum on you, with me in mind that you have 50 ppm of arsenic; m in os from a p ro x im a d to m e n t 300 ppm of ca lc io; m in os from a p ro x im a d to m e n te 200 ppm of the lo; m in os from a p ro x im a d to m e n t 50 ppm of coba lto; m in os from a p ro x im a d to m e n t 50 ppm of charge; m in os from a p ro x im a d to m e n t 300 ppm of iron; m in os from a p ro x im a d to m e n te 2% by weight of water; less than 50 p.m. to 50 ppm p lom o; m in os from a p ro x im a d to m e n t 50 ppm of m aga ness; m in os from a p ro x im a d to m e n te 0.2 ppm m ecu rio; m in os a p ro x im a d a m e n t 50 ppm m or libdene; m in os from a p ro x im a d to m e n te 1% by weight of n itro ge no; m in os from a p ro x im a d to m e n t 200 ppm of power; m in os from a p ro x im a d to m e n t 300 ppm of sodium; m in os from a p ro x im a d to m e n te 3% by weight of sugar; m in os of 50 ppm zinc; and / or m in os 700 ppm phosphorus ro.

En a lg u n o s asp ec tos , los a lcoh o le s g rasos, los es te res g rasos, las ce ra s y los h id ro ca rb u ro s p ro du c ido s segun las d ivu lg a c io n e s de este d o cum e n to , con tienen en tre a p ro x im a d a m e n te 50 % y a p ro x im a d a m e n te 90 % de carbono; en tre a p ro x im a d a m e n te 5 % y a p ro x im a d a m e n te 25 % de h id rogeno ; o en tre a p ro x im a d a m e n te 5 % y a p ro x im a d a m e n te 25 % de ox igeno . En o tros a sp e c to s de las rea liza c io nes , los a lco h o le s g rasos, los e s te re s grasos, las ceras y los h id ro ca rb u ro s p ro du c ido s com o se de sc rib e en este d o cum e n to , con tien en en tre a p ro x im a d a m e n te 65 % y a p ro x im a d a m e n te 85 % de ca rbono ; en tre a p ro x im a d a m e n te 10 % y a p ro x im a d a m e n te 15 % de h id rogeno ; o en tre a p ro x im a d a m e n te 10 % y a p ro x im a d a m e n te 20 % de oxigeno .

In some cases, the latter or the latter, the latter, the ce s and the id r d u r sp u du sp ro du c gone s according to the rules of this do cum in to, with between 50% and 50% carbon 90% carbon; from 5% to 5% and from 25% to 25% of hydrogen; or between 5% and 5% and at least 25% of oxygen. In some of the realities of the realities, those in the hobbies, the fatty ones, the waxes and the oils are the spheres of the commons. or I know about this in this year, with 65% of the time being 85% and 85% of carbon; 10% to 10% and 10% to 15% of hydrogen; or between 10% to 10% and 20% to 20% oxygen.

VI. Composiciones de combustible

SAW. Fuel compositions

C o m o se d e sve la en el p re se n te do cu m e n to , c ie rto s d e riv a d o s de ac ido s g ra so s fa b ric a d o s seg un los m e to d o s y las c o m p o s ic io n e s d e sc rito s en el p resen te do cu m e n to , poseen d ive rsa s ca ra c te ris tica s v e n ta jo sa s para su uso com o co m b u s tib le . Un e xp e rto en la m a te ria a p re c ia ra que, d e p e n d ie n d o de l p ro p o s ito p re v is to de l co m b u s tib le , d ife re n te s d e riva d o s de ac ido s g ra sos pueden te n e r v e n ta ja s en co m p a ra c io n con o tros de riva dos de ac ido s g rasos. P or e jem p lo , los d e riva dos de ac idos g ra sos ra m ifica d o s pueden s e r m as d e sea b le s com o co m b u s tib le s para a u to m o v ile s o co m o c o m p o n e n te s de c o m b u s tib le s pa ra a u to m o v ile s que es ta n de s tin a d o s a su uso en c lim a s frios. De m an e ra s im ila r, en d e te rm in a d a s ap lica c io ne s , puede s e r v e n ta jo s o p ro d u c ir un co m b u s tib le que este m as o m en os o x ig e n a d o o m as o m en os sa tu rado .

How do I know how to do it in the past, what is the place of interest in my life according to the methods and the composition of the rituals? In the case, they have a rich sven ta jo sa s for their common use as a bus. An example in the matter will be that, depending on what it is going to be, it will take two years of access. Next, they can keep you in touch with other people from both sides. For example, those of riva dos of acidic acids can be more than just as common as for combustion as combustion components. It is also clear that it is still useless in cold limes. On the other hand, in terms of practical applications, it may be helpful to go to a bus that is more or more in the oxidized or more in It's your name.

U tiliza n d o los m e todos de sc ritos en el p re sen te docum e n to , pueden p ro d u c irse co m b u s tib le s que com p ren dan d e riva d o s de ac ido s g ra sos re la tiva m e n te h o m o g e n e o s que, al m ism o tie m p o , te n g a n las ca ra c te ris tica s /cu a lid a d e s desea da s . D ichos co m b u s tib le s ba sad os en d e riva d o s de ac ido s g ra sos pueden ca ra c te riza rse po r la hue lla de ca rb o n o y po r su fa lta de im purezas , en com p a ra c io n con los co m b u s tib le s de riva dos del pe tro leo o el b iod iese l de riva d o de tr ig lice rid o s , ta m b ie n resu lta s e r una ven ta ja . Los co m b u s tib le s ba sados en de riva dos de ac idos g rasos pueden c o m b in a rse con o tros co m b u s tib le s o ad itivos de com b u s tib le para p ro d u c ir co m b u s tib le s que te n g a n las p ro p ieda de s deseadas .

Using the methods of sciences in the docum docum in all, you can leave as a bus that you can take away from two of the acquaintances. Let us, at the same time, deny you the most critical aspect of what you want to do. Such co-busses have been sad to see if they can continue to grow in the sense of purity and their fate of purities, as compared to With the two busses of Riva Two of the Petroleum, or the newspaper of Riva do de Tri lice ridos, it also turns out to be a veta ta. The cobus buses based on riva two of common acids can be combined with other tibial bus carriers for the bus to go with the bus that are denied you the desired expectations.

Los ho sp e d a d o re s de p ro du cc ion y los m e tod os de sve la d o s en el p re sen te d o cu m e n to pueden u tiliza rse para p ro d u c ir a c id o s g ra so s lib res y es te re s g rasos. En a lg u n o s asp ec tos , los h o sp e d a d o re s de p ro du cc ion y los m e tod os d e sve la d o s en el p re sen te d o cu m e n to pueden u tiliza rse para p ro d u c ir un titu lo o re n d im ie n to m as alto y /o m e jo rado de d e riva d o s de ac ido s grasos, in c luye nd o , po r e jem p lo , ac ido s g rasos lib res y /o e s te re s g rasos. En a lgu nos aspec tos , el po rcen ta je de ac ido s g rasos lib res en el p roducto p ro du c ido po r el h o sp e d a d o r de p roducc ion es de al m en os a p ro x im a d a m e n te 1 %, po r e jem p lo , de al m en os a p ro x im a d a m e n te 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 % , 15 % , 20 % o 25 % . En a lgu nos aspec tos , el po rcen ta je de es te res g rasos en el p ro du c to p ro du c ido po r el h o s p e d a d o r de p ro du cc ion es de al m en os a p ro x im a d a m e n te 50 %, po r e jem p lo , de al m en os a p ro x im a d a m e n te 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 % o 90 %. En a lgu nos aspec tos , la re lac ion de e s te re s g ra sos a ac idos g ra sos lib res en el p ro d u c to p roduc ido po r el h o sp e d a d o r de p ro du cc ion es de a p ro x im a d a m e n te 10:1, 9:1, 8:1, 7:1, 5:1, 2:1 o 1:1. En d e te rm in a d o s asp ec tos , el e s te r g ra so p ro d u c id o po r el h o s p e d a d o r de p ro du cc ion es d o d e ca n o a to de e tilo , tr id e c a n o a to de etilo , te tra d e c a n o a to de e tilo , p e n ta d e ca n o a to de etilo , c /s -9 -h e xa d e ce n o a to de etilo, h e xa d e ca n o a to de etilo, he p ta d e ca n o a to de etilo , c /s -11 -o c ta d e ce n o a to de etilo , o c ta d e ca n o a to de etilo, o co m b in a c io n e s de los m ism os. En o tros asp ec tos d e te rm in ado s , el e s te r g ra so p ro d u c id o po r la p ro du cc ion es el d o d e ca n o a to de m etilo , tr id e c a n o a to de m etilo , te tra d e c a n o a to de m etilo , p e n ta d e ca n o a to de m etilo , c/s-9-h e xa d e ce n o a to de m etilo , h e xa d e ca n o a to de m etilo , he p ta d e ca n o a to de m etilo , c /s -11 -o c ta d e ce n o a to de m etilo , o c ta d e c a n o a to de m e tilo , o co m b in a c io n e s de los m ism os. En c ie rto s a sp e c to s de re a liza c io n e s , el ac ido g ra so lib re p ro d u c id o po r el h o sp e d a d o r de p ro du cc ion es ac ido d o de can o ico , ac ido te tra d e ca n o ico , ac ido pen tad eca no ico , ac ido c /s -9 -h e xa d e ce n o ico , ac ido he xad eca no ico , ac ido c /s -11 -o c tade cen o ico , o c o m b in a c io n e s de los m ism os.

The past few days of protection and the two methods at the end of the year can be used for the purpose of going to each other. And this is true. In some cases, the hoped e o r ents of s p ro du cc ion and the methods of sve both in the p rst p e rd in which they can be used in order to go t o tu n re nd im ie n to higher and / ome jo rado de de riva two fatty acids, in c luye nd o, po re jem p lo, ac gone sg frogs libres y / o e s You are sure. In some respects, the power of the second century in the public sector is now at least 5 years old. 1%, po re jem p lo, of al m in os ap ro x im adamen te 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20% or 25%. In some respects, the power of these resources in the case of the host of the hospital is at least in the same period of time. Adamen te 50%, po re jem p lo, from at least in ap ap x x adamen te 55%, 60%, 65%, 70%, 75%, 80%, 85% or 90%. In some respects, the re ction of these will be followed by the acidic acids in the product du p roduc ted by the ho age of the p du duction is of ap ro x im adamen te 10: 1, 9: 1, 8: 1, 7: 1, 5: 1, 2: 1 or 1: 1. In te rm in ados asp ec tos, he will be able to support the host of the pipeline, where he will be an ethiopian, he will bring you Decathlon of ethyl, pen of ethyl acetate, c / s -9-he xa of ethyl acetate, hexane of ethyl ether, of ethyl ether, c / s -11 -octa of ethyl acetate, octa of ethyl ether, or co mb in the same way. In other countries of the world, he is responsible for the idea of being the ethyl pipeline, the ethyl canal of the ethyl group. , he is treated by ethyl, pen of alcohol, c / s-9- hex of ce ea to ethyl, he xa of cath of ethyl, he is Noe de m ethyl, c / s -11 -oc ta ce noa de m ethyl, oc ta decanoa de me tilo, or co mb in acions of the same. In c sa rto sa sp ec to s de re a liza c io nes, the ac i g ra so lib re p ro duc id o po r the ho sp age of p ro du cc ion is ac i do do can o ico , it has brought you from a non ico, it has been pen ead no ico, it has been c / s -9 -he xa from ce no ico, it has gone xca eca no ico, it has been c / s -11 -oc It is also central to the work of the same.

Los ho sp e d a d o re s de p ro du cc ion y los m e tod os de sve la d o s en el p re sen te d o cu m e n to pueden u tiliza rse para p ro d u c ir d ife ren te s p ro p o rc io n e s de ac idos g ra sos lib res y e s te re s g rasos. En a lgunos aspec tos , la p ro po rc ion de ac ido s g ra sos lib res en el p ro d u c to se pu ede m o d ifica r de a cu e rd o con los m e todos , las c o m p o s ic io n e s , los ve c to re s y las ce lu la s d e sc rito s en el p re se n te d o cu m e n to de ta l m an e ra que la p ro po rc ion es m a yo r o m e n o r en co m p a ra c io n con los es te res g ra sos que se p ro du cen . En c ie rto s asp ec tos re lac io na do s , la p ro po rc ion de e s te re s g ra sos en el p ro du c to ta m b ie n se puede m o d ifica r segun las d ivu lg a c io n e s de este docum e n to , de m odo que la p ropo rc ion sea m a yo r o m e n o r en com p ara c ion con los o tros p roductos, po r e jem p lo , los a c id o s g rasos libres, que se p roducen . En o tros asp ec tos d e te rm in ado s , el re n d im ie n to p ro po rc ion a l de l de riva d o de ac ido g raso con c ie rta s lo ng itude s de ca d e n a de ca rb o n o se pued e a u m e n ta r o d ism inu ir.

The past few years of the crisis and the methods of the two at the end of the year can be used for the purpose of going to the future. These acidic acids are free and he is safe. In some respects, the right to continue in the public can be modified from time to time with the methods, the compositions, I see them and I know that they are the rites in the world that the reason for this is that I am more or less comfortable with This is what you need to know. In this case, the reasons for this, the risk of these activities in the future can be modified according to the requirements of the law. acions of this document in any way that the p rpo rion is Most of me in com- pration with the other products, for example, the free offices, which are produced. In other countries of the world, the network of the private sector with the mainland with the main lines of the Canadian capital rb ono can also be ta rod ism inu ir.

A. Huella de carbono

A. Carbon footprint

Los de riva dos de ac idos g ra sos p ro du c ido s b io lo g ica m e n te rep rese n tan una nueva fu e n te de com b us tib les , ta les com o a lcoho les , d iese l y gaso lina . Los b io co m b u s tib le s fa b rica d o s de acu e rd o con los m e tod os y co m p o s ic io n e s d e sc rito s en este d o cu m e n to no se han p ro d u c id o hasta ah o ra a p a rtir de fu e n te s ren ova b le s y son nuevas c o m p o s ic io n e s de m a te ria . E stos nuevos co m b u s tib le s se pueden d is tin g u ir de los co m b u s tib le s de riva d o s de ca rb o n o pe tro qu im ico b a san do se en la dob le hu e lla iso to p ica de ca rbono . A d ic io n a lm e n te , la fuen te esp e c ifica de ca rb o n o de o rigen b io lo g ico (p o r e jem p lo , g luco sa fren te a g lice ro l) se puede d e te rm in a r m ed ia n te la dob le hue lla iso to p ica de ca rb o n o (vea se la P a te n te de E s tad os U n ido s N° 7 169 588, en pa rticu la r, de la col. 4, linea 31, a la col.

Those of riva dos de g acid are also responsible for a new era in the use of drugs, such as drinks, gas, and gas. . The two-year-old co-bus companies have been acquainted with the methods and co-ops of the rites in this period when they have not been able to do so until now. Appearing from the world is a new composition of materials. These new cobus buses can be found in the cobalt bus companies in Rio de Janeiro, which are based on the chemical in the basics of the main topic of ca rbono. At the end of the year, the special source of ca rb ono de o rigen b io lo gico (pore jem p lo, g luco sa fren te ag lice ro l) can be made in arm In this regard, I think it is the main topic of the ca rb ono (see the P eratio of E s tad os U n gone s No. 7 169 588, in pa rrticu la r, of the col 4, line 31, to col.

6, linea 8).

6, line 8).

Los de riva dos de ac ido s g rasos y los b io co m b u s tib le s aso c ia dos , los p ro du c to s qu im icos y las m ezc las , se pueden d is tin g u ir de sus hom o logo s p ro ced en te s de p ro du c to s p e tro qu im icos ba san do se en el 14C (fM) y en la dob le hue lla iso to p ica de carbono .

Those from Riva Dos de Ac Sag Rasos and the B Cobus Bus Associates, the Chemists and the Chemists, can be guided by their homosexuals. logo sp ro ced en te s of p ro du c to spe tro chemics based on 14C (fM) and on the dob is the topical carbon carbon.

Los de riva dos de ac ido s g ra sos de sc ritos en el p re sen te do cu m e n to tien en u tilidad en la p ro du cc ion de b io co m b u s tib le s y p ro du c to s qu im icos . Los nuevos p ro du c to s ba sad os en d e riva d o s de ac ido s g rasos p ro p o rc io n a d o s po r la p resen te d ivu lg a c io n se pueden d is tin g u ir ad ic io n a lm e n te b a san do se en la do b le hue lla iso to p ica de ca rb on o de aq ue llo s m a te ria le s de riva dos e xc lu s iva m e n te de fu e n te s pe tro qu im icas . La ca p ac idad de d is tin g u ir es tos p ro du c to s es b e ne fic iosa para el ras treo de es to s m a te ria le s en el com e rc io . P or e jem p lo , los c o m b u s tib le s o p ro du c to s q u im ico s que co m p re n d e n pe rfile s de iso to po s de ca rb o n o ta n to "n u e vo s" com o "v ie jo s " se pu ed en d is tin g u ir de los c o m b u s tib le s y p ro du c to s qu im ico s fa b rica d o s de m a te ria le s "v ie jo s". P o r tan to , los p re sen te m a te ria le s pueden se g u irse o "ra s tre a rse " en el co m e rc io o pueden id e n tifica rse en el co m e rc io com o un b io co m b u s tib le b a san do se en su perfil un ico. A de m as , o tros m a te ria le s c o m p e tid o re s pueden id e n tifica rse com o d e riva d o s b io lo g icos o d e riva d o s de una fu e n te pe troqu im ica .

Those from Riva Dos de Acdo Sg Ra SOS are from R Rites in the P re S te rn that they have a good time in the cooperation of the Cobus Bus. . The new p ro du c to s ba sad os en de riva dos de ac gone sos frogs p ro po rc io nados po r la p resen te d ivu lg acion can be given to go ad go to the lm na lm In the basics, the main topic of ca rb on or here is where you will be the only two of the two men and women in which you are interested in icas The capacity of d is tin g u go is that it is beneficial for the course of this period in the community. P o e jem p lo, the combus tib le sop ro du c to squ im ico s that also co r rnden pe rfile s iso to po s de ca rb ono ta n to "new vo s" com o "v ie jo s "can be found in the combustion of the combus tib le syp ro du c to s chemic s fa b rica two de ma te ria le s" v ie jo s ". For example, the parents may be able to keep or "move on" in the community or they can be identified in the community as a community. Cobus bus le ba san do se in its unique profile. On the other hand, other people may be able to identify themselves as being logical or directive of a peterchemistry.

En a lgu nos e jem p los , se fa b ric a una co m p o s ic io n de b ioco m bus tib le , que in c luye un de riva d o de ac ido g ra so que tie n e un v a lo r 813C de a p ro x im a d a m e n te -10 ,9 a a p ro x im a d a m e n te -15 ,4 , en do nde el de riva do de ac ido graso rep rese n ta al m en os a p ro x im a d a m e n te el 85 % de l m a te ria l de o rigen b io lo g ico (es decir, p ro ced en te de un recu rso ren o va b le ta l com o, po r e jem p lo , m a te ria le s ce lu lo s ico s y a zu ca re s ) en la com p os ic ion . En o tros e jem p los , la co m p o s ic io n de b io co m b u s tib le in c luye un de riva d o de ac ido g ra so que tie n e la fo rm u la :

In some of them, we have made a bus company, which includes a private area so that there is a 813C from ap ro x im adamen te -10, 9 aap ro x im adamen te -15, 4, where that of fatty acid riva do re m to m in ap ap x x adamen te 85% They are the only ones in which they are biologically (that is, they are in a given box, or they are good, but they are also important. s ico sya zu ca re s) in the community. In other countries, the co-p o s ic io n de b io co m b u s tib le includes a riva d o d a g ive so that it has the form:

X -(C H (R ))nC H 3

X - (C H (R)) nC H 3

en la que

in which

X = C H ³ , -C H ² O R ¹ ; -C (O )O R 2; o -C (O )N R 3R4;

X = CH ³ , -CH ² OR ¹ ; -C (O) OR 2; or -C (O) NR 3R4;

R = para cad a n, in d e p e n d ie n te m e n te ausen te , una H, o un g ru po a lifa tico in ferio r;

R = for cad a n, in d e p e n d ie n you are absent, an H, or a lifeline group in ferio r;

n = un nu m ero en te ro de a p ro x im a d a m e n te 8 a a p ro x im a d a m e n te 34, p re fe re n te m e n te un nu m ero en te ro de a p ro x im a d a m e n te 10 a a p ro x im a d a m e n te 24;

n = a number in ap ro x im adamen te 8 aap ro x im adamen te 34, p re faith re n ment a number in ap ro x im adamen te 10 aap ro x im adamen te 24;

R 1, R2, R3, R4 = se se le cc io n a n in d e p e n d ie n te m e n te de un H o un a lq u ilo in ferio r.

R 1, R2, R3, R4 = you will be told in the case of an H or a year in rio.

N o rm a lm e n te , cua nd o R es un g ru po a lifa tico in fe rio r, R rep rese n ta un g rupo a lqu ilo in fe rio r o a lq u e n ilo in fe rio r ram ificado , no ra m ifica d o o c ic lico . Los g ru po s R e je m p la re s inc luyen , sin lim itac ion , m etilo , isop rop ilo , isobu tilo , sec-butilo , c ic lope n ten ilo , y s im ila res . El de riva do de ac ido g ra so se ca ra c te riza ad em a s po r te n e r un v a lo r 513C de a p ro x im a d a m e n te -10 ,9 a a p ro x im a d a m e n te -15 ,4 , y el de riva d o de ac ido g raso rep rese n ta al m enos a p ro x im a d a m e n te el 85 % de l m a te ria l de o rigen b io lo g ico en la com p os ic ion . En a lgu nos e jem p los , el de riva do de ac ido g ra so en la co m p o s ic io n de l b io co m b u s tib le se c a ra c te riza po r te n e r una fra cc io n de l ca rb o n o m od e rn o (fM 14C) de al m en os a p ro x im a d a m e n te 1,003, 1 ,010 o 1,5.

N o rm a lm in te, when R is a lifeline group in faith r, R r re nt a g rupo a lqu ilo in fe rio roa lq uen ilo in fe rio branched, It is not my docical document. G ru po s R e je mp la re s inc include, without limitation, m ethyl, isopropyl, isobu tyl, sec- butyl, c iclope n ten-yl, and imsila. The riva do de ac ido g ra so se ca ra c te la riza ad em a s po r te ner a va lo r 513C of ap ro x im adamen te -10, 9 aap ro x im adamen te -15, 4 , and that of the private sector represents at least 85% of the logic of the logical rule in the com- pany. In some of them, the person in charge of life in the community will also be able to make a mistake of it. ca rb onom od e rn o (fM 14C) of al m in os ap ro x im adamen te 1,003, 1, 010 or 1.5.

B. Impurezas

B. Impurities

Los de riva dos de ac ido s g rasos p re pa rado s segun las d ivu lg a c io n e s del p resen te d o cum e n to , son u tiles com o co m p o n e n te s de, o para la fa b rica c io n de, b ioco m bus tib le s , a s i com o otros p ro du c to s q u im ico s in du s tria le s . Estos d e riva d o s de ac ido s g ra sos estan fa b ric a d o s d ire c ta m e n te a p a rtir de a c id o s g ra so s y no de l p ro ce sa m ie n to q u im ico de los tr ig lice rid o s . P o r con s ig u ie n te , los co m b u s tib le s y o tros p ro du c to s q u im ico s in d u s tr ia le s que com p re n d e n los d e riva d o s de ac ido s g ra so s d e sve la d o s a m en ud o con tie n e n m en os im p u reza s con las que n o rm a lm e n te se asoc ian , po r e jem p lo , los p ro du c to s de riva d o s de tr ig lic e r id o s ta le s com o co m b u s tib le s de riva dos de ace ites y g rasas veg e ta les .

Those of riva dos de arc sg rasos p re pa rado s in accordance with the rules of the priests are useful, they are useful as co-ops of, or for the famous manufacturer. n b, b ioco m bus tib le s, as well as others for the same in s du du tria le s. These riva dos de acido sg ra sos are fa b ric adosd ire c ta men te a te r away from a c id osg ra so s and not lp ro ce sa sa ie the chemist of the tr i lice rid os . For example, the co-buses are the only ones that have the benefits of the two of the two in the second century. Dosam in you or with time in which you are associated with those who are not associated with you, may not be associated with the two of three of three countries. It is the same as the two busts of oil and urban plants.

Los b io co m b u s tib le s p ro ced en te s de ac ido s g ra sos c rud os p re pa rado s segun las d ivu lg a c io n e s del p resen te d o c u m e n to (a n te s de m e z c la r el d e r iva d o de ac ido g raso con o tros co m b u s tib le s com o los c o m b u s tib le s ba sa d o s en pe tro le o ) con tienen m en os ca ta liza d o re s de tra n se s te r if ica c io n que el d iese l basado en pe tro leo u otro b iod iese l p ro d u c id o m ed ia n te una o m as e ta p a s de tra n s e s te r if ic a c io n . El d e riva d o de a c id o g ra so pued e c o n te n e r m en os que a p ro x im a d a m e n te 2 ,0 % , po r e jem p lo , m en os que a p ro x im a d a m e n te 1 ,5 % , 1 ,0 % , 0 ,5 % , 0 ,3 % , 0,1 %, 0 ,05 % o 0 % de un c a ta liz a d o r de tra n se s te r if ica c io n o una im p u reza resu lta n te de un c a ta liz a d o r de tra n se s te rif ica c io n . C o m o e je m p lo s no lim itan te s de ca ta liza d o re s de tra n se s te r if ica c io n se in c luyen ca ta liza d o re s de h id rox ido , ta le s com o N aO H , KO H y L iO H ; y c a ta liza d o re s ac idos, ta le s com o ca ta liza d o re s de ac ido m in e ra l y ca ta liza d o re s de ac ido de Lew is. C om o e je m p lo s no lim ita tivo s de c a ta liza d o re s e im p u reza s resu ltan te s de c a ta liza d o re s de tra n s e s te r if ic a c io n se in c luye n es tano , p lom o, m ercu rio , cadm io , c inc, titan io , c ircon io , ha fn io , boro, a lum in io , fos fo ro , a rse n ico , an tim on io , b ism uto , ca lc io , m ag ne s io , es tronc io , uran io , po tas io , sod io , litio, y co m b in a c io n e s de los m ism os.

The busses have a lot of knowledge in the areas of the city, according to the rules of the document that are documented (before mixing the project). Derivative of the common law with other co-buses, as well as the combusts, in two but it has more than enough to know that the day is based on a petrol or other binary ie lp ro duc id om ed ia Do not miss this step of work. That of riva do de c id og ra so may be important in that ap ro x im adamen te 2, 0%, po re jem p lo, m in which ap ap x x adamen te 1, 5%, 1, 0%, 0, 5%, 0, 3%, 0.1%, 0, 05%, or 0% of a trader of the work is not only an im pu reza resu The leader of a class is the leader in the field. How do I know what is not possible? What is the reason for the introduction of the right of way of life? or N aO H, KO H and L iO H; And it is the case of acidic acids, the most common type of acidic drugs and the type of Lewy acidity. C om oe je mp lo s no lim ita tive s de ca ta liza do re se im pu pra re s s resu ltan te s ca ca l l liza do re s de tra nses te r if ic ac io n se in c luye n es size, p lom o, m ercu rio, cadm io, c inc, titan io, c ircon io, ha fn io, boron, a lum in io, phosphorus, a rse nico, an tim on io, b ism uto, ca lc io, m ag ne s io, is tronc io, uran io, po tas io, sod io, lithium, and co mb in tions of the same.

Los b io co m b u s tib le s p ro ced en te s de ac ido s g ra sos c rud os p re pa rado s segun las d ivu lg a c io n e s del p resen te d o cu m e n to (a n te s de m e zc la r los d e riva d o s de a c id o s g ra so s con uno o m as co m b u s tib le s ) tie n d e n a te n e r un pun to de ge lifica c io n bajo, esp e c ia lm e n te cuando el p roduc to de riva d o de ac ido g raso com p ren de un e s te r e tilico C m ¹ o un e s te r e tilico C ¹⁸ : ¹ , en co m p a ra c io n con los pu n tos de ge lifica c io n de o tros t ip o s de b ioco m bus tib le s .

The busses have a great deal of interest in the areas of the city according to the decisions of the president of the country. The riva two of Ac id osg ra so s with one or more busses, also have a low gestation point, especially when the product is produced. from the private area of a public entity C m ¹ or a public service C ¹⁸ : ¹ , in comparison with the gestation points n o o tros t ip os de b ioco m bus tib le s.

De m an e ra s im ila r, los b io co m b u s tib le s p ro ced en te s de ac ido s g ra sos c rud os p re p a ra d o s segun las d ivu lg ac ion es de l p re sen te d o cu m e n to (an tes de m e z c la r e l(los ) d e riva d o (s ) de ac ido s g ra so s con uno o m as com b us tib les , com o los d iese l ba sad os en pe tro leo u o tros b iod iese l), con tien en m en os g lice ro l (o g lice rin a ) que los b ioco m bus tib le s fa b rica d o s con tr ig lice rid o s . El o los de riva dos de ac ido s g ra sos pueden co n te n e r m en os que a p ro x im a d a m e n te 2 ,0 % , po r e jem p lo , m en os que a p ro x im a d a m e n te 1 ,5 % , 1 ,0 % , 0 ,5 % , 0 ,3 % , 0,1 %, 0 ,05 % o 0 % en peso de g lice ro l.

In the same way, the buses with the tibial spreads are given in the same way according to the terms and conditions of the law. What do you think about (before mixing the rel riva do (s) of ac c a sg ra so s with one or more com b us tib les, com dsse l ba sad os en pe tro tro leo u tros b iod iese l), with me in os g lice ro l (og lice rin a) that the b ioco m bus tib le s fa b rica two with tr ig lice rid os. The two people in this area may know that in the case of 2, 0%, in which they may be 1, 5 %, 1, 0%, 0, 5%, 0, 3%, 0.1%, 0.05% or 0% by weight of g lice ro l.

Los b io co m b u s tib le s c rud os p ro ced en te s de los d e riva d o s de ac ido s g ra sos de l p resen te d o cu m e n to tam b ien con tien en m en os a lco h o l(e s ) lib re (s), (p o r e jem p lo , a lcoh o le s que se u tilizan para c re a r el es te r) que los b iod iese l fa b rica d o s con tr ig lice rid o s . Esto se debe en pa rte a la e fica c ia de la u tilizac ion de los a lcoh o le s po r parte de los h o sp e d a d o re s de p roducc ion de la p re sen te d ivu lg ac ion . P o r e jem p lo , el de riva do (o de riva d o s ) de ac ido s grasos puede co n te n e r m en os que a p ro x im a d a m e n te 2 ,0 %, 1,5 %, 1,0 %, 0 ,5 %, 0 ,3 %, 0,1 %, 0 ,05 % o 0 % en peso de a lcoh o l libre.

The b bus co tib bus sc rud os p ro ced in te riva's two of ac cg sg ra sos de lp resen te do te men also also have in me a lco hol (es) lib re (s), (pore jem p lo, to lcoh or s that are used to make this one) that the two countries have two with three licenses. This is due in part to the effectiveness of the use of the a few years or part of the hours of production of the production of the shareholder. P o jem p lo, that of riva do (or of riva dos) of fatty acids can be considered to be approximately 2, 0%, 1.5%, 1.0%, 0.5%, 0.3%, 0.1%, 0.05% or 0% by weight of free alcohol.

El b io co m b u s tib le p ro ced en te de los de riva dos de ac idos g ra sos de sve lado s , puede c a ra c te riza rse ad ic io n a lm e n te po r su ba ja co n ce n tra c io n de azu fre en co m p a ra c io n con el d ie se l d e riva d o de pe tro leo . El b io co m b u s tib le de riva d o de los d e riva d o s de ac ido s g ra sos en este d o cu m e n to pued e te n e r m en os que a p ro x im a d a m e n te 2 ,0 %, po r e jem p lo , m en os que a p ro x im a d a m e n te 1,5 %, 1,0 %, 0 ,5 %, 0 ,3 %, 0,1 %, 0 ,05 % o 0 % en peso de azu fre .

The bus company also welcomes you in those of the two major acid acids on the side, it can be added to you in the future. With the help of sugar in commpa ra tio with the leadership of petro leo riva. The bus company has the riva do of those of riva dos de acido sg ra you are in this do, where you can get on with it for about 2%, po re jem p lo , m in which ap ro x im adamen te 1,5%, 1,0%, 0, 5%, 0, 3%, 0.1%, 0.05% or 0% by weight of sugar.

C. Aditivos y composiciones de combustible

C. Fuel additives and compositions

Los a d itivo s de co m b u s tib le se u tilizan pa ra m e jo ra r el re n d im ie n to de un c o m b u s tib le o m otor. P o r e jem p lo , pueden u tiliza rse a d itivo s de co m b u s tib le para a lte ra r los p u n tos de c o n g e la c io n /g e lifica c io n , los p u n tos de e n tu rb a m ie n to , la lub ric idad , la v isco s id a d , la es ta b ilida d ox ida tiva , ca lidad de ign ic ion , los n ive les de oc ta no y los pun tos de ign ic ion . En los E s tados U n idos, to d o s los ad itivo s de co m b u s tib le deben e s ta r re g is tra d o s en la A g e n c ia de P ro tecc ion A m b ie n ta l. Los nom bres de los ad itivo s de co m b u s tib le y las co m p a n ia s que com e rc ia liza n los ad itivo s de co m b u s tib le es tan d isp o n ib le s al pub ico co n ta c ta n d o con la E P A o v is ita n d o el s itio w e b de la agenc ia . Un experto h a b itu a l en la m a te ria ap re c ia ra que los d e riva d o s de ac ido s g ra sos de sc ritos en el p re sen te d o cu m e n to pueden m e zc la rse con uno o m as a d itivo s de co m b u s tib le para im p a rtir una ca lida d deseada .

Collective assets are used for the benefit of a temporary loan. For example, they can be used as an addictive of the bus stop to freeze the freezing points, the points of your life, the points in your life. , the richness, the validity of the identity, the positive status, the ignition quality, the ocular levels and the ignition points. In the United States, all of the cooperative agents owe it to them in the A p e te ntio n of the P ro tection A m b ie n ta l. The names of the Cobus bus agents and the companies that the Cobus busses are so popular with the public as well as the public. c ta ndo with the EPA ov is ita the website of the agency. An expert who speaks in the ma ria ap re c ia ra will say that those of riva two of ac c a sg ra are those of r rites in the p r p r o te where they can zc to leave with one more ad ive It can be used to prevent a desired fall.

Los d e riva dos de ac ido s g ra sos de sc ritos en el p re sen te d o cu m e n to pueden fo rm u la rse en ad itivo s de com b us tib le ad ecu ad os , lo que m e jo ra el ren d im ien to de los co m b u s tib le s o m otores. P o r e jem p lo , los de riva d o s de ac idos g ra sos de sc rito s en el p resen te d o cu m e n to pueden fo rm u la rse com o m e jo ra d o re s de la lub ric idad , que im parten p ro p ieda de s de sea b le s , ta le s com o p ro te cc io n con tra el de sga s te a las pa rtes de l m otor. P o r co n s ig u ie n te , se p ro po rc io n a n com p o s ic io n e s ad itivas que com p ren den los de riva dos de ac ido s g ra sos p ro du c ido s segun las d ivu lg a c io n e s de l p resen te do cu m e n to . En o tro e jem p lo , los d e riva d o s de ac ido s g ra sos de sc rito s en el p re sen te d o cu m e n to pu ed en fo rm u la rse com o in h ib id o res de la corros ion .

Those from riva dos de acido sg ra are from sc rites in the p re sen te do how they can go on ad sive of com b us tib le ad ecu ad os, which makes me better REPRESENTATION OF BUS COMPANIES P o jem p lo, those of riva dos de g acid are sc rito s in the p penteent in which they can become the most common member of lub rity, which im They leave for the sake of sea b le s, such as the op t le with the sga s te to the parts of the grant. For the same reason, it is possible for the ad hoc commissions that include those of riva dos de ac ido sg ra sos p ro du c gone s according to the d ivu lg What are you doing? In o tro e jem p lo, those in riva dos de ac ido sg ra sos de sc rito s in the p re sen te do cu men to pu ed in fo rm u la rse com o in h ib id o res de la Ion corrosion.

Los d e riva d o s de ac ido s g ra sos d e sc rito s en el p re se n te d o cu m e n to pu ed en m e zc la rse con o tros c o m b u s tib le s ta le s com o uno o m as b iod iese l de riva dos de tr ig lice rid o s , d ive rso s a lcoh o le s , ta le s com o e ta no l y bu tano l, y p roduc tos d e riva d o s del pe tro leo , ta le s com o la ga so lina o el d iese l. En de te rm in a d a s c ircu n s ta n c ia s , se p roduce un de riva do de a c id o g ra so con un pu n to de ge lifica c io n ba jo , ta l com o un e s te r e tilico C ¹⁶ : ¹ o un e s te r e tilico C ¹⁸ : ¹. E ste de riva do de a c id o g ra so de pun to de ge lifica c io n ba jo puede m ezc la rse con uno o m as b iod iese l fa b rica d o s de tr ig lice rid o s pa ra re d u c ir el pun to de g e lif ica c io n de l c o m b u s tib le re su lta n te cu a n d o se c o m p a ra con un co m b u s tib le que con tien e so lo uno o m as b io d ie se l fa b rica d o s de tr ig lice rid o s . De m an era s im ila r, un de riva d o de ac ido graso , com o un e s te r e tilico C ¹⁶ : ¹ o e s te r e tilico un C ¹⁸ : ¹ , puede m ezc la rse con un d iese l p ro ced en te de pe tro le o pa ra p ro p o rc io n a r una m ezc la que con te n g a al m en os a p ro x im a d a m e n te , y con frecue nc ia , m as que a p ro x im a d a m e n te , 5 % en peso de b iod iese l. En a lgu nos e jem p los , la m ezc la de com b u s tib le in c luye al m en os a p ro x im a d a m e n te 10 % , 15 % , 20 % , 30 %, 40 %, 50 % y 60 % en peso de l de riva d o de ac ido graso.

Those from Riva Dos de Acido Sg Ra SOS from Sc Rito S in the P re se n te do cu men to Pu ed in me zc la rse with o tros combus tib le s ta le s com o one or more b iod iese l de riva dos de tr ig lice rid os, d ive rso sa lcoh o le s, ta le s com oe ta no ly bu tano l, yp roduc tos riva dos del pe tro leo, ta le s com o la ga So lina or the day l. In te rm in adasc ircu ns ta nc ia s, a riva do de c a o id ra is produced with a low ge ge ation, a comm C ¹⁶ : ¹ or one is te C ¹⁸ : ¹ . This riva do de a c id og ra so de ge lifica c io n io r u can be left with one or more b iod iese l fa b rica two de tr ig lice rid os pa ra re duc Going to the point of genetics with the bus company will show you how much it compares with a bus that has only one or more of the two factories. de tr ig lice rid os. In the same way it is im ila r, one of the riva do of fatty acid, like a C ¹⁶ : ¹ or t he re til a C ¹⁸ : ¹ , can be left with a d See that you have a personal license for a period of time, and that you often have more time, and often more than just a year. im adamen te, 5% by weight of b iod iese l. In some years, the most important part of the bus in the world is 10%, 15%, 20%, 30%, 40%, 50% and 60 % by weight of the fatty acid riva do.

En a lg u n o s asp ec tos , la co m p o s ic io n de c o m b u s tib le pu ede c o m p re n d e r adem as un co m b u s tib le s in te tico . C u a lq u ie r co m b u s tib le s in te tico ob te n id o de carbon , gas na tu ra l o b iom asa , puede u tiliza rse ad e cu a d a m e n te . En un aspecto ad ic iona l, el co m b u s tib le s in te tico co m p re n d e un co m b u s tib le b a sad o en F isch e r-T ro p sch , un co m b u s tib le b a sad o en B erg ius , un co m b u s tib le b a sad o en M ob il, un co m b u s tib le b a sad o en K arrick , o una com b in ac io n de e llos . En o tros a sp e c to s ad ic io na le s , el co m b u s tib le s in te tico co m p re n d e un co m b u s tib le ba sad o en ca rbon a liqu idos (co m b u s tib le ba sad o en C TL, Coal-To-Liquids), un co m b u s tib le ba sado en gas a liqu idos (co m b u s tib le basado en G TL, Gas-To-Liquids), un co m b u s tib le ba sado en b iom asa a liqu id os (co m b u s tib le ba sado en BTL), un co m b u s tib le ba sad o en ca rb on y b iom asa a liqu idos (co m b u s tib le basado en C B TL, Coal and Biomass-To-Liquids), o una com b in ac io n de los m ism os. En un a sp e c to e jem p la r, el co m b u s tib le s in te tico c o m p re n d e un co m b u s tib le ba sado en F ische r-T ro psch . La can tidad de co m b u s tib le s in te tico en la com p o s ic io n de co m b u s tib le de sve la d a en el p resen te d o cu m e n to puede s e r de a p ro x im a d a m e n te 5 % a a p ro x im a d a m e n te 90 %, de a p ro x im a d a m e n te 5 % a a p ro x im a d a m e n te 80 %, de a p ro x im a d a m e n te 5 % a a p ro x im a d a m e n te 70 %, de a p ro x im a d a m e n te 5 % a a p ro x im a d a m e n te 60 % o de a p ro x im a d a m e n te 5 % a a p ro x im a d a m e n te 50 %.

In some cases, the combustion of tibus can also include a more complex bus. Each of the coal companies with coal coal, gas in your lobe, can be used ad hoc. In an aditional aspect, the bus is also in co-op with a bus that is sad or in F isch e rT ro p sch, a bus is tib le ba sad or in B erg ius , a bus tib le ba sad or in M ob il, a bus bus tib le ba sad or in K arrick, or a com bus in them. In other countries, the co-bus company has a co-bus with a cobus bus that is sad or liquid-based (cobus bus). in C TL, Coal-To-Liquids), a cobus bus based on gas in liquids (co bus based on G TL, Gas-To-Liquids), a bus bus based on liquids handle liquids (co bus tib le based on BTL), co bus bus le le sad or in charge and drink liquids (co bus bus based on CB TL, Coal and Biomass-To- Liquids), or a comm in ation of the same. In one year, the bus company is composed of a bus company based in Forsche rT ro psch. The amount of cobus busses is in technical in the bus cobus bussiness group only the day in which the budget can be of approximately 5% a year. x im adamen te 90%, from ap ro x im adamen te 5% aap ro x im adamen te 80%, from ap ro x im adamen te 5% aap ro x im adamen te 70%, from ap ro x im adamen te 5% aap ro x im adamen te 60% or ap ro x im adamen te 5% aap ro x im adamen te 50%.

En un a sp ec to de te rm in ado , se puede p re p a ra r una com p o s ic io n de b io co m b u s tib le que in c luya al m enos a p ro x im a d a m e n te 5 %, 10 %, 20 %, 30 %, 40 %, 50 %, 60 %, 70 %, 80 %, 85 %, 90% o 95% de un de riva do de ac ido g ra so que in c luya una ca d e n a de ca rb o n o que es 8: 0, 10: 0, 12: 0, 14: 0, 14: 1, 16: 0, 16: 1, 18: 0, 18: 1, 18: 2, 18: 3, 20: 0, 20: 1, 20: 2, 20: 3, 22: 0, 22: 1 o 22: 3. D ichas c o m p o s ic io n e s de b io co m b u s tib le pueden in c lu ir a d ic io n a lm e n te al m en os un ad itivo se le cc io n a d o de un ad itivo que reduce el pun to de e n tu rb a m ie n to que puede d is m in u ir el pun to de e n tu rb a m ie n to a una te m p e ra tu ra m e n o r de a p ro x im a d a m e n te 5 °C o m e n o r de a p ro x im a d a m e n te 0 °C; un te n s io a c tivo ; una m ic ro em u ls io n ; al m en os a p ro x im a d a m e n te 5 % , 10 % , 15 % , 20 % , 30 %, 40 %, 50 %, 60 %, 70 %, 80 %, 85 %, 90 % o 95 % de co m b u s tib le d iese l de tr ig lice rid o s ; una ga so lin a ob ten ida de pe tro leo ; o un co m b u s tib le d iese l de pe tro leo .

In a year of development, it is possible to prepare a bus com b ation that starts at least 5%, 10%. , 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, or 95% of a rural area so that a chain of channels is included rb ono which is 8: 0, 10: 0, 12: 0, 14: 0, 14: 1, 16: 0, 16: 1, 18: 0, 18: 1, 18: 2, 18: 3, 20: 0, 20: 1, 20: 2, 20: 3, 22: 0, 22: 1 or 22: 3. These compositions of the bus can also be added to you in At the same time, an ad- tive will be added to an ad- tive that reduces the point of your rb am ie n that the point of your rb am ie n can go one smaller than a minimum of 5 ° C or less than 0 ° C; a te n s io a c tivo; a m em ro em u ls io n; to m in os ap ro x im adamen te 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90% or 95% of co mbus tib le le ie l de tr ig lice rid os; a ga so lin a ob ten ida de pe tro leo; or a co b b s s tib le d iese l pe pe troro leo.

En a lgu nos aspec tos , la com p o s ic io n de co m b u s tib le que com p ren de los es te res g ra sos p ro du c ido s segun los m e todos, los ve c to re s , las ce lu las y las c o m p o s ic io n e s de la p resen te inven c ion com p ren den a d e m a s uno o m as a d itivo s para co m b u s tib le s d iese l. Los ad itivo s ad e cu a d o s son, de sea b le m en te , aq u e llo s que p ro po rc ion an un re n d im ie n to m e jo ra d o pero ta m b ie n co m p a tib ilid a d con los c o m p o n e n te s en la co m p o s ic io n del co m b u s tib le y d isp o s itivo s que estan aso c ia d o s n o rm a lm e n te con m o to res d iese l. C o m o e jem p los ilu s tra tivo s de o tros a d itivo s de co m b u s tib le a d e cu a d o s se inc luyen m e jo ra d o re s de ign ic ion o m e jo ra d o re s de l nu m ero de ce tano , de te rge n tes , d isp e rsa n te s , ag en te s a n tide sga s te , m o d ifica d o re s de l ind ice de v isco s id a d , m o d ifica d o re s de la fricc ion , m e jo ra d o re s de la lub ric idad , es ta b ilizan te s , an tiox idan tes , in h ib id o res de co rro s ion , b ioc idas, d e sa c tiva d o re s de m e ta les y pe qu en as c a n tid a d e s de o tros ad itivo s opc io na les , inc luyendo , sin lim itac ion , a n tie sp u m a n te s y se llado res . En asp e c to s pa rticu la res , a m en ud o se an ad en m e jo ra d o re s de ign ic ion o m e jo ra d o re s del nu m ero de ce ta no para m e jo ra r el ren d im ien to de l m o to r d iese l. Los m e jo rad o re s e je m p la re s del ind ice de ce ta no in c luye n n itra to de 2 '-e tilh e x ilo y o tros n itra tos de a lqu ilo . Los m e jo rad o re s del ind ice de ce ta no pueden an a d irse a una co m p o s ic io n de co m b u s tib le en una can tida d que es de a p ro x im a d a m e n te 0,01 % en peso a a p ro x im a d a m e n te 1,0 % en peso, po r e jem p lo , de a p ro x im a d a m e n te 0 ,05 % en peso a a p ro x im a d a m e n te 0 ,5 % en peso, basado en el peso to ta l de la co m p o s ic io n de com b ustib le .

In some respects, the com- bustion of the bus company that understands what they are doing according to the methods, I see them, What are the compositions and compositions of the present invention that also allow one or more admissions for cobus busses? The ad sivents are, of course, that is why they have a better income but they also have a good idea With the components in the bus company, they are associated with the company in terms of how they are. How do I get the most illustrious traffic of the cobus busts also include the best ignition ignition of the number of the number of ce tano, de te rge n tes, d isp e rsa n te s, ag en te san tide sga s te, modification of the ice id of v isco s id ad, modified mod s of friction I am better at lub ric idad, it is also important, an antiox idan tes, in h ib id o res de cor r s sion, b ioc idas, de sa c tiva do re s de me They are also eligible for additional options, including, without limitation, even if they are known. In this regard, the country has been added to the improvement of the number of ignition of the number of cents not to improve the ren im lien to lmo to rd iese l. The m e jo rad o re s e je m p re s of the ice index of ce ta no in c luye n n itra to 2 '-e tilh e x ilo y o tros n itra coughs of aquilo. The improvements in the ice index cannot be added to a bus company in a quantity that is approximately 0.01% by weight. aap ro x im adamen te 1.0% by weight, po re jem p lo, from ap ro x im adamen te 0.05% by weight aap ro x im adamen te 0.5% by weight, based on the weight to This is the com com usbib le com.

En d e te rm in a d o s aspec tos , en la com p o s ic io n de co m b u s tib le que c o m p re n d e el e s te r g ra so p ro du c ido segun las p re sen te s d ivu lg a c io n e s , pueden in c lu irse d ive rso s de te rg e n te s y /o d isp e rsa n te s para a so c ia r y d is p e rs a r o e lim in a r d e p o s ito s d a n in o s de las p iezas de l m o to r d iese l. Los d e te rg e n te s a d e cu a d o s co m p re n d e n n o rm a lm e n te una cab eza p o la r que com p ren de una sa l m e ta lica de un co m p u e s to o rg an ico a c id o y una la rga co la h id ro fo ba . C o m o e jem p los de de te rg e n te s se in c luye n sa les b o ra tad as de ca rb on a to , sa les bo ra tad as de su lfona to , que estan p re fe ren te m en te sob re b a sa d a s . V eanse , po r e jem p lo , las pa ten tes de E s tados U n idos n.° 4 744 920 y 4 965 003. C om o e je m p lo s de d isp e rsa n te s se inc luyen , sin lim itac ion , d isp e rsa n te s ca rb ox ilicos , d isp e rsa n te s de succ in im ida , d isp e rsa n te s de am in a y d isp e rsa n te s de M ann ich . V ea nse , po r e jem p lo , las p a te n te s de E E .U U . n.° 3 172 892, 3 438 757, 3 980 569 y 6 165 235. Los d isp e rsa n te s pueden e s ta r p re sen te s en la com p o s ic io n de co m b u s tib le en una can tida d de a p ro x im a d a m e n te 0,01 % en peso a a p ro x im a d a m e n te 0,1 % en peso, po r e jem p lo , de 0 ,03 a a p ro x im a d a m e n te 0 ,05 % en peso, ba sado en el peso to ta l de la co m p o s ic io n de com b ustib le .

In terms of specific aspects, in the com- bustion of the bus that he understands, he is ruled according to the circumstances of his life, they can go d ive rso s de te rg in te sy / od isp e rsa n te s for a so c ia ryd is pe rs aroe lim in ardep or s ito sda n in os de lmo to p rd see l. Those of you who are aware of how you are comm rm a lm in which you have a capacity for the distribution of a technical salary of a collective cost or a rga co la h id ro fo ba. How do you think you are in love with your friends, who are your parents, who are happy to see you? You are re ba sa das. See, po re jem p lo, the patents of E s tados U n idos No. 4 744 920 and 4 965 003. C om oe je mp lo s of d isp and rsa n s are included, without lim itation, d isp e rsa n te s ca rb ox ilicos, d isp e rsa n te s de succ in im ida, d isp e rsa n te s de am in ayd isp e rsa n te s de M ann ich See, for example, the parents of E U. U U. No. 3 172 892, 3 438 757, 3 980 569 and 6 165 235. The d isp e rsa n s s can be also present in the bus com bustion in a can tida d ap ap x im adamen te 0.01% by weight aap ro x im adamen te 0.1% by weight, po re jem p lo, from 0.03 aap ro x im adamen te 0.05% by weight , based on the total weight of the comm com com usb le.

En d e te rm in a d o s a sp ec tos , a la c o m p o s ic io n de co m b u s tib le pu eden an a d irse a g e n te s a n tid e sg a s te , inc luyendo , po r e jem p lo , sa les m e ta lica s de d ih id ro ca rb il d itio fos fa to , para p ro p o rc io n a r be ne fic ios ta n to de an tid e sg a s te com o a n tiox idan tes . V ease , po r e jem p lo , la p a ten te de E E .U U . n.° 5 898 023.

In te rm in adosa sp ec tos, the composition of co bus tib can be added to leave in the future, including you, you will be the only one from d ih id ro ca rb il d itio fos fa to, for p ro po rc io nar be ne fic ios ta n to de an tid e sg as they com oan thiox idan tes. See, po r e jem p lo, the p a ten e of E E. U U. No. 5,898,023.

En asp e c to s pa rticu la res , la can tida d de m e jo ra d o r de la lub ric ida d en la com p o s ic io n de com b u s tib le puede v a r ia r de a p ro x im a d a m e n te 1 ppm a a p ro x im a d a m e n te 50 000 ppm , po r e jem p lo , de a p ro x im a d a m e n te 10 p p m a a p ro x im a d a m e n te 20 000 ppm , o de a p ro x im a d a m e n te 25 ppm a a p ro x im a d a m e n te 10 000 ppm . Los e jem p los no lim ita n te s de m e jo rad o re s de la lu b ric ida d in c luye n e s te re s y ac ido s g rasos , que pu ed en o no s e r los m ism o s que los p ro d u c id o s segun los m e tod os d e sc rito s en es te docum e n to .

In the case of the cattle, the amount of improvement of the public in the composition of the bus may vary from about 1 ppm to 1 ppm. aap ro x im adamen te 50 000 ppm, po re jem p lo, of ap ro x im adamen te 10 ppmaap ro x im adamen te 20 000 ppm, or ap ro x im adamen te 25 ppm aap ro x im adamen te 10,000 ppm The jem p los no lim ita n te s de me jo rad o re s de la lu b ric ida d in c luye n e s te re s y ac gone sos rasos, which may or may not be the same That the P ducts are according to the methods of science in this document.

En asp ec tos pa rticu la res , la can tidad de e s ta b ilizan te s , que m e jo ra la e s ta b ilida d de a lm a ce n a m ie n to de la com p o s ic io n de com b us tib le , puede v a r ia r de a p ro x im a d a m e n te 0,001 % en peso a a p ro x im a d a m e n te 2 % en peso, po r e jem p lo , de a p ro x im a d a m e n te 0,01 % en peso a a p ro x im a d a m e n te 1 % en peso, basado en el peso to ta l de la co m p o s ic io n de com b us tib le . Un e s ta b iliza n te e je m p la r es una am ina p rim a ria de a lq u ilo te rc ia rio .

In the case of the country, the amount of this work, which will improve the life of the commendation of the community of com Also, it can vary from ap ro x im adamen to 0.001% by weight aap ro x im adamen te 2% by weight, po re jem p lo, from ap ro x im adamen te 0.01 % by weight aap ro x im adamen te 1 % by weight, based on the total weight of the comm com com usb tib tib le . A sta b iliza n te e je mp la r is an am ina p rim a ria de a lq u ilo te rc ia rio.

Los a n tio x id a n te s im p iden la fo rm a c io n de depo s itos de go m a en los co m p o n e n te s del s is te m a de com b u s tib le d e b ido a la ox id a c io n de los co m b u s tib le s en el a lm a ce n a m ie n to y /o inh iben la fo rm a c io n de co m p u e s to s de pe rox ido en c ie rta s com p o s ic io n e s de com b us tib le . La can tidad de a n tio x id a n te s puede v a r ia r de a p ro x im a d a m e n te 0,001 % en peso a a p ro x im a d a m e n te 5 % en peso, po r e jem p lo , de a p ro x im a d a m e n te 0,01 % en peso a a p ro x im a d a m e n te 1 % en peso, ba sad o en el peso to ta l de la co m p o s ic io n de com b ustib le .

The years of identity have been the idea of the deposit of government agencies in the com- posters of the bus system because it has gone to the oxidization of The busses in the year to date and / or in part were the forum for commuting in the region of com com tib le. The amount of the year may vary from ap ro x im adamen to 0.001% by weight aap ro x im adamen te 5% by weight, po re jem p lo, from ap ro x im adamen te 0.01% by weight aap ro x im adamen te 1% by weight, ba sad or in the total weight of the comm com com us.

Los in h ib id o res de co rro s ion p ro tegen los m e ta les fe rro so s en los s is te m a s de m ane jo de com b us tib le , ta le s com o tu b e ria s y ta n q u e s de a lm a ce n a m ie n to , de la co rro s ion . T a m b ie n se sab e que c ie rto s in h ib id o res de co rro s ion im p arten lub ric ida d ad ic iona l, y com o ta le s son p a rticu la rm e n te a d e cu a d o s cua nd o se de se a una lu b ric idad ad ic iona l. El in h ib id o r de co rro s ion puede e s ta r p resen te en la com p os ic ion de co m b u s tib le en una can tida d de a p ro x im a d a m e n te 0,001 % en peso a a p ro x im a d a m e n te 5 % en peso, po r e jem p lo , de a p ro x im a d a m e n te 0,01 % en peso a a p ro x im a d a m e n te 1 % en peso, b a sad o en el peso to ta l de la co m p o s ic io n de com bustib le .

In h ib id ores de cor rro s ion p ro tete me fe les les rro so s in the most common ways of using it, it is common to you and your family nques de a lm a ce nam ie n to, of the cor sion. It is also known that what is in charge of the taxation of the tax administration, and how they are part of the law in which All of them are given to a lucrative ad ic iona l. In h ib id or de cor r s sion may be also in the com bustion of the tibi le in an ap rox of ap ro x im adamen te 0,001% by weight aap ro x im adamen 5% by weight, po re jem p lo, of ap ro x im adamen te 0.01% by weight aap ro x im adamen te 1% by weight, ba sad or in the total weight of the comm ic ic com bustib le.

Los b ioc id as se u tilizan pa ra c o m b a tir el c re c im ie n to m ic ro b ia n o en la co m p o s ic io n de com b us tib le , que pueden es ta r p re sen te en la co m p o s ic io n de com b u s tib le a una con cen tra c ion de a p ro x im a d a m e n te 0,001 % en peso a a p ro x im a d a m e n te 5 % en peso, po r e jem p lo , de a p ro x im a d a m e n te 0,01 % en peso a a p ro x im a d a m e n te 1 % en peso, b a sad o en el peso to ta l de la co m p o s ic io n de com b ustib le .

The b ioc id as used to fight the c re c i cn i n to m ic ro b ia not in the commposi of com b us tib le, which can be ta rp re sen te in the COMPOSITIONS OF THE COMBUSTION OF A BUS WITH A CENTRO TION OF APPROX IMMEDIATE 0.001% BY WEIGHT AAPROX IM Adamen T 5% BY WEIGHT, BY PEOPLE OF APRO X IM Adamen te 0.01% by weight aap ro x im adamen te 1% by weight, ba sad or in the total weight of the comm us com.

Los d e sa c tiva d o re s de m e ta les sup rim en los e fe c to s ca ta litico s que tien en a lgu nos m e ta les , pa rticu la rm e n te el cob re , sob re la ox idac ion de com b ustib le , que puede es ta r p re sen te s en la com p os ic ion de co m b u s tib le en una ca n tid a d de a p ro x im a d a m e n te 0,001 % en peso a a p ro x im a d a m e n te 5 % en peso, po r e jem p lo , de 0,01 % en peso a a p ro x im a d a m e n te 1 % en peso, b a sad o en el peso to ta l de la com p o s ic io n de com b ustib le .

Those of sa c tiva do re s de me ta les sup rim in the e c e c t e ca l ta s ta lts that have in me some of them, pa rrticu la rm en te el cob re, sob re la ox idac Com b ion, which may be also present in the bus combustion in a capacity of appraisal 0.001% by weight a year or month 5% by weight, po re jem p lo, of 0.01% by weight aap ro x im adamen te 1% by weight, ba sad or in the total weight of the com com ustib le.

A d e m a s , pueden an a d irse m e jo ra d o re s de v isco s id a d , que son n o rm a lm e n te m a te ria le s po lim e rico s de pesos m o le cu la re s p ro m ed io en nu m ero de a p ro x im a d a m e n te 5 000 a a p ro x im a d a m e n te 250 000, y m o d ifica d o re s de fr icc io n , que son no rm a lm e n te co m p u e s to s o rg a n o -m o lib d e n o que con tien en azu fre , en pe qu en as can tidades . T a m b ie n pueden an a d irse in h ib id o re s de espum a , que inc luyen n o rm a lm e n te po lim e ro s de m e ta c rila to de a lqu ilo o po lim e ro s de d im e til s ilic io , a la com p o s ic io n de com b us tib le en una can tida d m e n o r de a p ro x im a d a m e n te 10 ppm . A s im ism o , pueden an ad irse se lla d o re s para g a ra n tiza r un se llad o e la s to m e rico ad ecu ad o e im p e d ir que en la co m p o s ic io n de co m b u s tib le pueda in c lu irse un m al se llad o p rem aturo .

In addition, they can add to the improvement of their identity, which are not limited to pesos, but they are also limited to pesos. ed io in ap ro x im adamen te 5,000 aap ro x im adamen te 250 000, and modified fr fr icc io, which are not rm a lm in te c co pts to so rg year - Mo lib deno than with blue sugar, in small quantities. They can also be added in Spain, which include no rm a lm in the po l e r s of the ta me r la rila to de lqu ilo o po lim e ro s de It is only possible to make up 10 ppm of less than 10 ppm. In addition, they can be added to be called s to guarantee a call to the rich ad ecuador and to ask that in the community of Tibet Co You can start a call by calling rem rem.

Ejemplos

Examples

Los s ig u ie n tes e je m p lo s ilus tran la g e n o m a n ip u la c io n de h o sp e d a d o re s de p ro du cc ion para p ro d u c ir de riva dos e sp e c ifico s de ac idos g rasos. En las f ig u ra s se ilu s tran ru tas b io s in te tica s im p lica d a s en la p roducc ion de de riva dos de ac ido s grasos.

The evidence is that the genoman ip is the home of the federal government for the military and two of the acidic acids. g satin. In the f u r s, there is an interest in the technical and political aspects in the production of fatty acids.

P o r e jem p lo , La FIG. 3 es un d iag ram a de la ru ta FAS que rep rese n ta las e n z im a s d ire c ta m e n te im p lica da s en la s in tes is de ac il-A C P . P ara a u m e n ta r la p ro du cc ion de d e riva dos de ac idos g rasos, com o ceras, e s te re s grasos, a lco h o le s g ra sos e h id ro ca rbu ros , una o m as de las en z im a s de sc ritas en este d o cum e n to pueden so b re e xp re sa rse o m u ta rse pa ra re d u c ir la in h ib ic ion po r re tro a lim e n ta c io n , con el fin de a u m e n ta r la ca n tid a d de a c il-A C P p roduc ida . A d ic io n a lm e n te , e n z im a s que m e tab o lizan los p ro du c to s in te rm e d io s para fa b ric a r p ro du c to s ba sados en ac ido s no g ra sos (por e jem p lo , rea cc io nes se cu n d a ria s ) pueden d e le c io n a rse o a te nua rse fu n c io n a lm e n te pa ra a u m e n ta r el flu jo de ca rb on o a tra ve s de la ruta de b io s in te tica ac ido s g ra sos (FA S ). En los s ig u ie n tes e jem p los , se de scriben m uch os h o sp e d a d o re s de p ro du cc ion que se han m o d ifica d o para a u m e n ta r la p ro du cc ion de ac ido s grasos.

P ore jem p lo, FIG. 3 is a day of the FAS route that represents the implicit enzymes in the synthesis of ac-A-CP. In order to increase the risk of de riva two of common acids, such as waxes, these are fatty, so that they are the same as the ID, which is one of The areas of sc ritas in this do cum en to can be taken to be omitted to be injected in order to limit this, with The end of the year is the increase in ac-A CP CP roduc. A d ic io na lm en te, enzymes that tab or liza the p ro du c to s in te rm ed io s for fa b ric arp ro du c to s based on ac i s not g ra sos ( For example , the reactions will be taken into account) they may read or continue to be available in order to increase the flow of money or through time. s of the route of b io s in te tica ac ido sg ra sos (FA S). In the next few years, many of them have been described as having been modified to increase the rate of access to the situation. fatty

Las FIG. 4 y 5 rep rese n tan ru tas b ios in te ticas que pueden g e n o m a n ip u la rse para p re p a ra r es te res g rasos y a lco h o le s grasos, re sp ec tiva m en te . La tra n s fo rm a c io n de cada sus tra to (p o r e jem p lo , a ce til-C o A , m a lon il-C o A , ac il-A C P , ac ido g ra so y ac il-C o A ) en cada p ro du c to (p o r e jem p lo , ace til-C o A , m a lon il-C o A , ac il-A C P , ac ido graso , ac il-C oA , a ld e h id o s g rasos , es te re s g ra so s y a lco h o le s g ra so s ) pu ede re a liza rse u tiliza n d o va rio s p o lip e p tid o s d ife re n te s que son m ie m bros de las c la ses de en z im a s ind icadas.

FIG. 4 and 5 re n b e nts so basic routes that they can be used to make sure that they are fatty and fatty, re fl ective. The work of each of their deals (for example, to C-A or C, to L-C or A, to C-CP, to G-1 so and to see) C or A) on each p ro du c to (for example, ace til-C or A, ma lon il-C or A, ac il-A CP, fatty acid, ac il-C oA, a ld eh id osg rasos, this re sg ra so sya lco ho le sg ra so s) i can re a liza rse u use the rio spo lip e p tid osd ife re n te s which are members of the c the ses of in indicated indices.

Los s ig u ie n te s e jem p los de sc rib en m ic ro o rg a n ism o s que se han g e n o m a n ip u la d o o que pueden g e n o m a n ip u la rse para p ro d u c ir a lco h o le s g rasos, e s te re s g ra sos e h id ro ca rb u ro s esp ec ificos .

I know that those of sc ribs in m icro or rg an isms that have genoman ip u doo that can genoman ip u rse for p ro duc go to lco ho le sg satos, he s re sg ra sos eh id ro ca rb u ro s esp ec icos.

EJEMPLO 1. CONSTRUCCION DE HOSPEDADORES DE PRODUCCION

EXAMPLE 1. CONSTRUCTION OF PRODUCTION HOSPEDADORES

Un h o sp e d a d o r de p ro du cc ion e je m p la r es LS 9001. LS9001 se p ro du jo m o d ifica n d o C41 (D E 3 ) a p a rtir de la s o b re e xp re s io n (S a in t B eaus ine , F ranc ia ) a la in ac tiva c ion de l gen fad E (a c il-C o A d e sh id rog en asa ).

A ho sp age of p ro du cc ion e je mp la r is LS 9001. LS9001 is p ro du jo modified C41 (DE 3) off of the sop re e xp re s io n (S a in t B eaus ine, F ranc ia) to the initiative of the gene fad E (ac il-C or A of sh id rog in handle).

En resum en , la cepa in ac tiva da fadE de E. coli se p re pa ro u tilizan do los ce b a d o re s Y a fV _ N o tI e Iv ry_O l para a m p lif ic a r a p ro x im a d a m e n te 830 pb de fad E en d irecc ion 5' y los ceb ad o res L p ca f ol y Lp ca R _ B a m para a m p lif ica r a p ro x im a d a m e n te 960 pb de fadE en d irecc ion 3'. P ara c re a r una con s tru cc io n para la de le c ion en m arco del gen fadE com p le to , se u tilizo P C R con sup e rp os ic ion . La con s tru cc io n de de le c ion fad E se c lo no en el p lasm id o pK O V 3 se n s ib le a tem p e ra tu ra , que co n te n ia un gen sacB para la se lecc io n po r co n tra s te y se rea lizo una de lec ion c ro m o so m ica de fadE segun el m e tod o de L in k e t al., J. B act. 179 :6228 -6237 , 1997. La cep a resu lta n te no fue capaz de d e g ra d a r ac ido s g ra sos y a c il-C o A g rasos. En el p resen te d o cu m e n to es ta cepa in ac tiva da se de no m in a E. coli (D E 3, FadE).

In resume in the ac tive in strain E. coli fadE da is p re pa ro or tilizan do re ce Saturday And fV s _ No tI and Iv for ry_O l ic amp lif Arap ro x im adamen like 830 pb of fad E in direction 5 'and the ceb ad o res L p ca f ol and Lp ca R _ B am for amp amp ica rap ro x im adamen te 960 bp of fadE in direction 3'. P ara c re ar one with the structure for reading in the arc of the gene In addition , PCR was used with suppression. The fad E le c tion con c tio n is c in the p p id o pK OV 3 is your temper, which has a sacB gene for the se lesson for each other and one of the most important lessons of fadE according to the method of L in ket al., J. B act. 179: 6228-6237, 1997. The summary artist was not able to give a sg ra ac i sg ra sos yac il-C o A gos ce. In the present, this strain is not active in E. coli (DE 3, FadE).

O tra cepa de de lec ion FadE, M G 1655, se con s tru yo segun los p ro ce d im ie n to s desc ritos po r D a tse nko et al., P N A S (U S A ), 97: 6640 -6645 (2000 ), con las m o d ifica c io n e s que se d e sc rib e n a co n tin u a c io n . Los dos c e b a d o re s u tiliza d o s para c re a r la de le c ion fue ron :

Another strain of lesson FadE, MG 1655, is answered according to the criteria given by r n a tse nko et al., PNAS (USA), 97: 6640-6645 (2000), with the modifications that were made of sc rib ena co n tin uac io n. The two priming re used to read the lesson were:

Del-fadE-F: 5 ’-A A A A A C A G C A A C A A T G T G A G C T T T G T T G T A A T T A T A T T G T A A A C A T A T T G A T T C C G G G G A T C C G T C G A C C ; (S E Q ID N O :69 ) y

Del-fadE-F: 5 '-AAAAACAGCAACAATGTGAGC TTTGTTGTAATTATATTGTAA ACATATTGATTCCGGGGATCC GTCGACC; (SEQ ID NO: 69) and

Del-fadE-R: 5 ’-A A A C G G A G C C T T T C G G C T C C G T T A T T C A T T T A C G C G G C T T C A A C T T T C C T G T A G G C T G G A G C T G C T T C (S E Q ID N O :70).

Del-fadE-R: 5 '-AAACGGAGCCTTTCGGCTCCG TTATTCATTTACGCGGCTTCA ACTTTCCTGTAGGCTGGAGCT GCTTC (SEQ ID NO: 70).

C a d a uno de los c e b a d o re s D e l-fa d E -F y D e l-fad E -R , co n tie n e n 50 ba ses de h o m o lo g ia con el gen fadE de E. coli y se u tiliza ro n para a m p lif ic a r el case te de re s is te n c ia a ka n a m ic in a del p lasm id o pK D 13 po r P C T com o se describe . El p ro du c to de la P C R resu ltan te se u tilizo para tra n s fo rm a r ce lu la s M G 1655 de E. coli e le c tro co m p e te n te s que con tien en pK D 46. Las ce lu las se in du je ron p re v ia m e n te con a ra b inosa d u ran te 3-4 ho ras com o de sc rib e Datsenko, c itad o a n te rio rm en te . D e spues de 3 ho ras de c re c im ie n to en un m ed io S O C a 37 °C, las ce lu las se sem b ra ro n en p laca s de a g a r Lu ria que con ten ian 50 pg /m l de kan am ic ina . Las co lo n ia s res is te n tes se a is la ron de spu es de una in cu b a c io n d u ra n te la noche a 37 °C. La in te rrup c ion del gen fadE se con firm o en a lgu nas de las co lo n ia s m ed ia n te am p lif ica c io n po r P C R u tiliza n d o los fadE-L2 y fadE-R1, que se d isen a ron para f la n q u e a r el gen fadE .

Each one of the primers is D e l-fa d E-F and D e l-fad E-R, with 50 basic classes with the fadE gene of E. coli and Use it for the purpose of the case in which you will be able to use it as described. The result of the PCR was that they were used for the purposes of the MG 1655 of E. coli and that they have in pK D 46. The That is the reason why they have been with you at 3-4 hours, as well as sc rib e Datsenko, where they laughed at you. After 3 hours of rework in a SOC medium at 37 ° C, the semesters will be seized in the agar agar shells that had 50 pg. / ml of kan am ic ina. The most important factors are the rum after one night, at 37 ° C. The incorporation of the fadE gene was signed in some of the co-operative countries with PCR or using the fadE-L2 and fadE-R1, that rum was designed to identify the fadE gene.

fadE-L2 5 ’-C G G G C A G G T G C T A T G A C C A G G A C (S E Q ID N O :71); y

fadE-L2 5 '-CGGGCAGGTGCTATGACCAGG AC (SEQ ID NO: 71); and

fadE -R 1 5 ’-C G C G G C G T T G A C C G G C A G C C T G G (S E Q ID N O :72 )

fadE -R 1 5 ’-C G C G G C G T T G A C C G G C A G C C T G G (S E Q ID N O: 72)

D e sp u e s de c o n firm a r la d e le c io n a p ro p ia d a de fadE, se u tilizo una c o lo n ia pa ra re tira r el m a rc a d o r K m Ru tiliza n d o el p la sm id o pC P 20. La cepa resu lta n te se d e n o m in o M G 1655 (AfadE).

In accordance with the signature of the National Law of FadE, a co-operation was used for the shooting of the principal and the use of the law or pC P 20. The summary strain is called MG 1655 ( AfadE).

Los ho sp e d a d o re s con el gen fad E d e le c io n a d o se som e tie ro n a a jus tes ad ic io na le s . S e in tro du jo un p lasm ido que llevab a los cua tro genes que e ran re sp o n sa b le s de la ac tiv idad de la a c e til-C o A c a rb o x ila sa en E. coli (accA, accb accC y acd, N u m eros de R e g is tro de G en B ank: N P _ 414727 , N P _417721 , N P _ 417722 , N P _ 416819 , EC 6.4.1.2 ). Los genes accABCD se c lo n a ro n en dos e ta pas com o op e ro n e s b ic is tron icos en los s itio s Ncol/HindIII y Ndel/AvrlI de pA C Y C D uet-1 (N o vag en , M ad ison , W I), y el p lasm id o resu ltan te se de no m in o pA S 004.126. C om o a lte rna tiva , el h o s p e d a d o r de p ro du cc ion se g e n o m a n ip u lo para e x p re s a r accABCD de Lactobacillus plantarum.

The pastors with the fad E gene of the family fell asleep just ad na io na le s. It is a problem that will lead to the four genes that are responsible for the activity of acetyl-C or A ca rb ox ila sa in E. coli ( accA, accb accC and acd, Numbers of R eg is of G in B ank: NP_ 414727, NP_417721, NP_ 417722, NP_ 416819, EC 6.4.1.2). The accABCD genes can be found in two stages of communication in the Ncol / HindIII and Ndel / Avrl sites of pA CYCD uet-1 (N o vag en, M ad ison, WI), and the pasm id o resu ltan te de no m in o pA S 004.126. C om a lte rna tiva, the host of the pipeline genoman ip u lo to exp la sar accABCD of Lactobacillus plantarum.

O tra s m o d ifica c io n e s que se inc luye ron en un h o sp e d a d o r de p ro d u cc io n in c luye ron lo s igu ien te : s o b re e xp re s io n de aceEF (que cod ifica el co m p o n e n te E1p d e sh id ro g e n a sa y el co m p o n e n te E 2p d ih id ro lip o a m id a a c iltra n s fe ra sa de los com p le jos de p iruva to y 2 -o xo g lu ta ra to de sh id ro g e n a sa ); y fabH/fabD/fabG/acpP/fabF (que cod ifica FA S ) de E. coli, Nitrosomonas europaea (A T C C 19718), Bacillus subtilis, Saccharomyces cerevisiae, Streptomyces spp, Ralstonia, Rhodococcus, Corynebacteria, Brevibacteria, Mycobacteria y le vad u ras o lea g in osa s . De m an era sim ilar, se g e n o m a n ip u la ro n ho sp e d a d o re s de p ro du cc ion para e xp re sa r accABCD (que co d ifica la ace til C o A ca rb ox ilasa ) de Pisum savitum. S in em bargo , cuando el h o sp e d a d o r de p ro du cc ion ta m b ie n p ro du c ia bu tano l, se de scub rio que era m en os de se a b le e x p re s a r el ho m o lo g o de Pisum savitum.

The work that is included in a period of age in the world is as follows: on account of aceEF (which codifies the co they show the E1p of sh id ro gena sa and the co mpone te E 2p d ih id ro lip oam id aac iltra ns fe ra sa of the com p le jos de p iruva to y 2 -o xo g lu ta ra to de sh id ro gena sa); and fabH / fabD / fabG / acpP / fabF (encoding FA S) of E. coli, Nitrosomonas europaea (ATCC 19718), Bacillus subtilis, Saccharomyces cerevisiae, Streptomyces spp, Ralstonia, Rhodococcus, Corynebacteria, Brevibacteria and Mycodbacteria, Mycod u flush or read g in osa s. In a similar way, they are generated as a source of protection for the action (which codes the acetyl C or A ca rb ox ilasa) of Pisum Savitum S in em bargo, when ho sp age of p ro du cc ion ta mb ie np ro du c ia bu tano l, it was scub laughed that it was more than a few days ago. from Pisum savitum.

En a lg u n o s h o sp e d a d o re s de p ro du cc ion , los ge ne s se in ac tiva ron o a te n u a ro n u tiliza n d o el m e tod o de Link, e t al., J. B ac te rio l. 179 :6228 -6237 , 1997. Los genes que se in a c tiva ro n o a te nua ron , in c lu ye ro n g p sA (que co d ifica la sng lice ro l 3 -fo s fa to d e sh id ro g e n a sa b ios in te tica , n.° de reg is tro N P _ 418065 de G e n B ank , EC: 1.1.1.94); ldhA (que cod ifica la lac ta to de sh id ro g e n a sa , n.° de reg is tro N P _ 415898 de G en B ank, EC: 1.1.1.28); pflb (que cod ifica la fo rm a to a ce tiltra n s fe ra sa 1, n.° de reg is tro P 09373 de G en B ank, EC: 2.3.1.54 ); adhE (que cod ifica la a lcoho l de sh id ro g e n a sa , n.° de reg is tro C A A 47743 de G en B ank, EC: 1.1.1.1 , 1.2.1.10); pta (que co d ifica la fo s fo tra n sa ce tila sa , n.° de reg is tro N P _ 416800 de G e n B ank , EC: 2.3.1.8 ); poxB (qu e co d ifica la p iruva to ox ida sa , n.° de reg is tro N P _ 415392 de G e n B ank , EC: 1.2.2.2 ); ackA (que cod ifica la a ce ta to c inasa , n u m ero de reg is tro N P _ 416799 de G en B ank, EC: 2.7.2.1 ), y co m b in a c io n e s de los m ism os.

In some recent years, the people have been active in the process of Link, et al., J. B ac tio rio l . 179: 6228-6237, 1997. The genes that were activated in the past did not stop you, in which the sng lice ro l 3 -fo s fa to de sh id ro gena sa b ios in te tica, reg. no. NP _ 418065 of G in B ank, EC: 1.1.1.94); ldhA (which codifies the lacta to of sh id ro gena sa, reg.number NP _ 415898 of G in B ank, EC: 1.1.1.28); pflb (which encodes the photo rm a to a ce tiltra ns fe ra sa 1, reg. no. P 09373 of G in B ank, EC: 2.3.1.54); adhE (coding for the code l of sh id ro gena sa, reg.number CAA 47743 of G in B ank, EC: 1.1.1.1, 1.2.1.10); pta (which codifies the fo s fo tra n sa ce tila sa, reg.number NP _ 416800 of G in B ank, EC: 2.3.1.8); poxB (what does the p uuva to ox ida sa, reg. no. NP _ 415392 of G in B ank, EC: 1.2.2.2); ackA (which encodes the cease to c inase, registration number NP _ 416799 of G in B ank, EC: 2.7.2.1), and changes in the same months.

De m a n e ra s im ila r, para la de le c ion de fadE, se in tro du jo la m u ta c io n P lsB [D 311 E ] en LS9001 para a te n u a r plsB. E sta m u tac ion d ism inu yo la can tidad de ca rb on o de sv ia do a la p ro du cc ion de fo s fo lip id o s . S e creo un a le lo que c o d ific a b a P ls B [D 311 E ] re e m p la za n d o e l co d o n G A C pa ra el a sp a rta to 311 con un cod on G A A para el g lu tam a to . El a le lo a lte ra d o se p re pa ro po r s in te s is g e n ica y el a le lo de tip o s ilve s tre p lsB c ro m o so m ico se ca m b io po r el a le lo plsB [D 311 E ] m u tan te u tiliza n d o el m e tod o de Link et al. (vea se an te rio rm en te ).

In a similar way, for the legacy of FADE, the P lsB [D 311 E] in LS9001 was introduced in LS9001 to continue plsB. This is the in m ation of the amount of money from the source of the funds. S e created a what the code for P ls B [D 311 E] represents the GAC code for the year to 311 with a GAA codon for the g tam tam to . The person with whom he or she is po e r is the only one and the person with the most important type of experience is also allowed to change the year. plsB [D 311 E] mu so you use the method of Link et al. (see rm an te rio en te).

EJEMPLO 2. MODIFICACIONES EN HOSPEDADORES DE PRODUCCION

EXAMPLE 2. MODIFICATIONS IN PRODUCTION HOSPEDERS

P ara la e xp res ion de d ive rsa s p ro te in a s que se u tilizan en la s in te s is de d e riva d o s de ac ido s grasos, se con s tru ye ro n los s ig u ie n te s p lasm idos . Las co n s tru cc io n e s se p re pa ra ron u tiliza n d o m e tod os e s ta n d a r de b io lo g ia m o lecu la r. T odo s los genes c lo n a d o s se pus ie ron ba jo el con tro l de p ro m o to re s in du c ib les po r IP TG (p o r e jem p lo , un p ro m o to r de T7, un p ro m o to r tac o un p ro m o to r lac).

For the purpose of the ive rsa sp ro te sins so that they are used in the syndicate of fatty acids, they are based on the differences You splashed. The co-ventors can be used for the use of the standard of payment. All the genes with which they were put under the control of the country in which the IP TG (for example, a month of T7, a p ro mo to r tac o a p ro mo to r lac).

El gen 'tesA (gen de tio e s te ra s a A, nu m ero de reg is tro N P _ 415027 de G en B ank, sin se cu e n c ia lid e r (S E Q ID NO: 31 ) (C ho y C ronan, J. B iol. C hem ., 270 :4216 -9 , 1995, EC: 3.1.1.5 , 3.1.2.-)) de E. coli, se c lono en un v e c to r pE TD uet-1 d ige rido con Ndel/AvrlI (el v e c to r pE TD uet-1 de sc rito en el p re sen te do cu m e n to esta d ispo n ib le en N ovagen , M ad ison , W I). Los genes que cod ifica n t io e s te ra sa s (T E ) ve g e ta le s de tipo FatB de Umbellularia californica, Cuphea hookeriana y Cinnamonum alcanfor (N u m e ro s de R e g is tro de G enB ank: U cF a tB 1 = A A A 34215 , C h F a tB 2 = A A C 49269 , C h F a tB 3 = A A C 72881 , C cF a tB = A A C 49151 ) se c lo na ron in d iv id u a lm e n te en tre s ve c to re s d ife ren te s : (i) pE T D ue t-1 d ig e rid o con NdeI/AvrII; (ii) p B lu e s c rip t KS+ d ig e rid o con XhoI/HindIII (S tra ta g e n e , La Jo lla , CA, para c re a r p ro te inas de fus io n la cZ ::T E N -te rm ina les ); y (iii) pM A L -c2 X d ige rido con XbaI/HindIII (N e w E ng land Lab, Ipsw ich , M A ) (p a ra c re a r fu s io n e s malE::TE n -te rm ina les ). El gen fadD (que cod ifica la a c il-C o A s in ta sa ) de E. coli se c lo no en un d e riva d o de pC D F D ue t-1 d ig e rid o con NcoI/HindIII, que c o n te n ia el gen acr1 (a c il-C o A red uc tasa ) de Acinetobacter baylyi A D P 1 de n tro de sus s itio s NdeI/AvrII.

The gen 'tesA (uncle gene is te ra sa A, registration number NP _ 415027 of G in B ank, without being a leader (SEQ ID NO: 31) (C ho and C ronan , J. B iol. C hem., 270: 4216-9, 1995, EC: 3.1.1.5, 3.1.2.-)) of E. coli, was extended in a neighborhood of TD TD-1 d Rigid with Ndel / AvrlI (the neighbor to the TD TD-1 of the Rite in the presence of the public in New Mexico, M ad ison, WI). The genes that codify nt io is te ra sa s (TE) see gels of the FatB type of Umbellularia californica, Cuphea hookeriana and Cinnamonum camphor (Numbers of R eg is of G enB ank: U cF a tB 1 = AAA 34215, C h F a tB 2 = AAC 49269, C h F a tB 3 = AAC 72881, C cF a tB = AAC 49151) were reported in d iv id ua lm in te between s ve c to re sd ife ren te s: (i) pE TD ue t-1 d ig e rid o with NdeI / AvrII; (ii) p B lu esc rip t KS + d ig e rid o with XhoI / HindIII (S ta ta gene, La Jo lla, CA, for c re arp ro f lation lines cZ :: TEN -te rm ina les); and (iii) pM AL-c2 X d ige rido with XbaI / HindIII (N ew E ng land Lab, Ipsw ich, MA) (pa ra c re ar fu s io nes malE :: TE n -te rm ina les) . The fadD gene (which encodes the ac il-C or A s in ta sa) of E. coli was named in a pC DFD riva do ue t-1 d ig e rid o with NcoI / HindIII, which with the acr1 gene (ac il-C or A red uc rate) of Acinetobacter baylyi ADP 1 from one of its NdeI / AvrII sites.

La Tabla 7 p ro p o rc io n a un resu m en de los p lasm id os ge n e ra d o s para c re a r va rio s ho sp e d a d o re s de p roducc ion e jem p la re s .

Table 7 p ro p o rc io n a summary of the p lasm id os ge n e ra d o s for c r a r va rio s ho sp e d a d o re s of p roduction e jem p la re s.

T l 7: R m n l mi iliz n l h r r i n

T l 7: R m n l mi iliz n l h r r i n

Un exp erto en la m a te ria a p rec ia ra que para o b te n e r cep as s im ila re s pueden u tiliza rse d ife ren te s p lasm id os y m o d ifica c io n e s ge no m icas .

An expert in the matter would say that in order to obtain them, they can be used if they can be used as a model and can be used to modify genetics.

Los plasmidos de expresion seleccionados contenian replicones y marcadores de resistencia a antibioticos compatibles para producir un sistema de expresion de cuatro plasmidos.

The selected expression plasmids contained replicons and compatible antibiotic resistance markers to produce a four plasmid expression system.

En algunos aspectos de realizaciones, LS9001 puede cotransformarse con: (i) cualquiera de los plasmidos que expresan TE; (ii) el plasmido que expresa fadD, que tambien expresa acrl; y (iii) el plasmido de expresion de ester sintasa.

In some aspects of embodiments, LS9001 can be co-transformed with: (i) any of the plasmids expressing TE; (ii) the plasmid expressing fadD, which also expresses acrl; and (iii) the ester synthase expression plasmid.

Como sera obvio para un experto en la materia, cuando LS9001 se induce con IPTG, la cepa resultante producira concentraciones mas altas de alcoholes grasos a partir de fuentes de carbono tales como glucosa.

As will be obvious to one skilled in the art, when LS9001 is induced with IPTG, the resulting strain will produce higher concentrations of fatty alcohols from carbon sources such as glucose.

EJEMPLO 3. PRODUCCION DE ALCOHOL GRASO EN LA CEPA DE E. C O L IR E C O M B IN A N T E

EXAMPLE 3. PRODUCTION OF ALCOHOL FAT IN THE CEPA DE E. COL IR ECOMB IN ANTE

En un hospedador de produccion, se produjeron alcoholes grasos expresando de manera exogena un gen de tioesterasa y un gen de acil-CoA reductasa. De manera mas especifica, los plasmidos pCDFDuet-1-fadD-acr1 (acil-CoA reductasa) y pETDuet-1-'TesA (tioesterasa) se transformaron en la cepa LS9001 de E. coli y los transformantes correspondientes se seleccionaron utilizando placas LB complementadas con espectinomicina 100mg/l y carbenicilina 50 mg/l. Cuatro transformantes de LS9001/pCDFDuet-1-fadD-acr1 se inocularon independientemente en 3 ml de un medio M9 complementado con carbenicilina 50 mg/l y espectinomicina 100 mg/l. Las muestras que contenian los transformantes se cultivaron a 25 °C en un agitador (agitacion a aproximadamente 250 rpm) hasta que alcanzaron una DO600 de 0,5. A continuacion, se transfirieron 1,5 ml de cada muestra a un matraz de 250 ml que contenia 30 ml del medio M9 descrito anteriormente. El cultivo resultante se cultivo a 25 °C en un agitador hasta que alcanzo una DO600 de entre 0,5-1,0. Despues, se anadio IPTG a una concentracion final de 1 mM. El cultivo celular continuo durante 40 horas.

In a production host, fatty alcohols were produced exogenously expressing a thioesterase gene and an acyl-CoA reductase gene. More specifically, plasmids pCDFDuet-1-fadD-acr1 (acyl-CoA reductase) and pETDuet-1-'TesA (thioesterase) were transformed into E. coli strain LS9001 and the corresponding transformants were selected using supplemented LB plates with spectinomycin 100mg / l and carbenicillin 50mg / l. Four transformants of LS9001 / pCDFDuet-1-fadD-acr1 were independently inoculated into 3 ml of an M9 medium supplemented with 50 mg / l carbenicillin and 100 mg / l spectinomycin. Samples containing the transformants were grown at 25 ° C on a shaker (shaking at approximately 250 rpm) until they reached an OD600 of 0.5. Next, 1.5 ml of each sample was transferred to a 250 ml flask containing 30 ml of the M9 medium described above. The resulting culture was grown at 25 ° C on a shaker until it reached an OD600 between 0.5-1.0. Then, IPTG was added to a final concentration of 1 mM. Cell culture continued for 40 hours.

Despues, las celulas se centrifugaron y se sedimentaron a 4000 rpm. El sedimento celular se suspendio en 1,0 ml de metanol. Despues, se mezclaron 3 ml de acetato de etilo con las celulas suspendidas, seguido de la adicion de 3 ml de H2O. A continuacion, la mezcla se sometio a utrasonido durante 20 minutos. La muestra resultante se centrifugo a 4000 rpm durante 5 minutos. Despues, la fase organica (la fase superior), que contenian el(los) alcohol(es) graso(s), se sometio a analisis de GC/Ms . El titulo de alcohol total (incluido el tetradecanol, hexadecanol, hexadecenol y octadecenol) fue de aproximadamente 1-10 mg/l. Cuando una cepa de E. coli que llevaba solo vectores vacios, se cultivaba en las mismas condiciones y siguiendo el mismo protocolo, se obtenia un titulo de alcoholes grasos de solo 0,2-0,5 mg/l.

Then, the cells were centrifuged and sedimented at 4000 rpm. The cell pellet was suspended in 1.0 ml of methanol. Then, 3 ml of ethyl acetate was mixed with the suspended cells, followed by the addition of 3 ml of H2O. Then, the mixture was subjected to utrasonide for 20 minutes. The resulting sample was centrifuged at 4000 rpm for 5 minutes. Then, the organic phase (the upper phase), which contained the fatty alcohol (s), was subjected to GC / Ms analysis. The total alcohol titer (including tetradecanol, hexadecanol, hexadecenol and octadecenol) was approximately 1-10 mg / l. When an E. coli strain that carried only empty vectors, was grown under the same conditions and following the same protocol, a fatty alcohol titer of only 0.2-0.5 mg / l was obtained.

1. Produccion de la muestra de biodiesel # 23-30

1. Production of biodiesel sample # 23-30

La muestra de biodiesel # 23-30 ("muestra # 23-30") se produjo mediante cultivo en biorreactor de una cepa de E. coli (C41 DE3 AfaaE AfabR 'TesA fadD adplws) genomanipulada para producir esteres grasos. Para la expansion del cultivo se utilizo un protocolo de inoculo de dos fases. La primera fase consistio en la inoculacion de un medio de LB 50 ml (complementado con carbenicilina 100 pg/l y espectinomicina 100 pg/l) en un matraz de agitacion deflector de 250 ml con un vial de 1 ml de material congelado de la cepa de E. coli de produccion de ester. Este matraz de semilla se incubo a 37 °C durante aproximadamente 7 horas (DO600 final = 4,5, pH 6,7), despues de lo cual se transfirieron 3 ml del cultivo primario a cada uno de tres matraces deflectores de 2 l que contenian medio F1 minimo tamponado 350 ml que tambien contenia carbenicilina 100 pg/l y espectinomicina 100 pg/l. El tampon del matraz de agitacion utilizado fue Bis-Tris propano a una concentracion final de 200 mM (pH 7,2). Estos matraces secundarios se incubaron a 37 °C durante aproximadamente 18 horas (DO600 final = 12, pH 5,5) y los contenidos se utilizaron para inocular tres biorreactores de 14 l con un volumen inicial de 6,5 litros de medio F1 minimo tamponado despues de la inoculacion. Estos biorreactores tambien contenian carbenicilina 100 pg/l y espectinomicina 100 g/l.

The biodiesel sample # 23-30 ("sample # 23-30") was produced by bioreactor culture of a strain of E. coli (C41 DE3 AfaaE AfabR 'TesA fadD adplws) genomanipulated to produce fatty esters. For the expansion of the culture, a two-phase inoculum protocol was used. The first phase consisted in the inoculation of a 50 ml LB medium (supplemented with 100 pg / l carbenicillin and 100 pg / l spectinomycin) in a 250 ml deflector shake flask with a 1 ml vial of frozen material of the strain of E. coli ester production. This seed flask was incubated at 37 ° C for approximately 7 hours (final DO600 = 4.5, pH 6.7), after which 3 ml of the primary culture was transferred to each of three 2L deflector flasks that contained 350 ml minimum buffered F1 medium which also contained 100 pg / l carbenicillin and 100 pg / l spectinomycin. The buffer of the stirring flask used was Bis-Tris propane at a final concentration of 200 mM (pH 7.2). These secondary flasks were incubated at 37 ° C for approximately 18 hours (final DO600 = 12, pH 5.5) and the contents were used to inoculate three 14 l bioreactors with an initial volume of 6.5 liters of buffered minimum F1 medium after inoculation. These bioreactors also contained 100 pg / l carbenicillin and 100 g / l spectinomycin.

Estos biorreactores de 14 l se cultivaron inicialmente a 37 °C, y los niveles de oxigeno disuelto se mantuvieron al 30 % de saturacion, utilizando los bucles en cascada de agitacion y enriquecimiento de oxigeno. El pH de la mezcla de fermentacion se mantuvo a 7,2, utilizando H2SO41 M y gas amoniaco anhidro. Cuando se agoto la carga de glucosa original de 5 g/l en el medio basal, se inicio un suministro de nutrientes en cada biorreactor que consistia principalmente en glucosa al 43 % (p/v). Despues, la velocidad de suministro de la solucion de glucosa se ajusto manualmente durante el ciclo de fermentacion para mantener la glucosa residual a un valor bajo (pero no nulo) durante el ciclo de fermentacion. Cuando la DO600 de los cultivos llego a un valor de 30, los cultivos se indujeron con una concentracion final de IPTG de 1 mM. En este punto de induccion, la temperatura de cultivo del biorreactor se redujo a 30 °C, y al cultivo se anadieron aproximadamente 15 ml/l (basandose en un volumen de 6,5 a 7 litros) de etanol y en todo momento se controlo mediante HPLC. Para mantener las concentraciones residuales a aproximadamente 20 ml/l, se anadio etanol adicional periodicamente a los biorreactores. Los contenidos de los biorreactores se recogieron despues de aproximadamente 60 horas de cultivo, con aproximadamente 10 l del caldo recogido de cada uno de los tres biorreactores.

These 14 l bioreactors were initially grown at 37 ° C, and dissolved oxygen levels were maintained at 30% saturation, using cascade loops of oxygen agitation and enrichment. The pH of the fermentation mixture was maintained at 7.2, using H2SO41 M and anhydrous ammonia gas. When the original glucose load of 5 g / l in the basal medium was depleted, a nutrient supply was initiated in each bioreactor consisting mainly of 43% glucose (w / v). Then, the supply rate of the glucose solution was manually adjusted during the fermentation cycle to keep the residual glucose at a low (but not zero) value during the fermentation cycle. When the OD600 of the cultures reached a value of 30, the cultures were induced with a final IPTG concentration of 1 mM. At this induction point, the bioreactor culture temperature was reduced to 30 ° C, and approximately 15 ml / l (based on a volume of 6.5 to 7 liters) of ethanol was added to the culture and at all times it was controlled by HPLC. To maintain residual concentrations at approximately 20 ml / l, additional ethanol was added periodically to the bioreactors. The contents of the bioreactors were collected after approximately 60 hours of culture, with approximately 10 l of the broth collected from each of the three bioreactors.

Estos caldos recogidos se combinaron y se extrajeron con un volumen equivalente de acetato de etilo con agitacion a te m p e ra tu ra am b ie n te du ran te dos horas. D espues, los ex trac to s de ca ldo se ce n trifu g a ro n (3 500 rpm , 30 m in u to s ) para se p a ra r las capas liqu idas, seg u ido de la e lim in ac ion de la capa o rg an ica para su po s te rio r p ro ce sa m ie n to . El ace ta to de e tilo se e lim in o cas i p o r c o m p le to (< 0 ,3 % res idu a l, segun lo d e te rm in a d o po r G C /F ID ) de la capa o rg a n ica u tilizan do eva p o ra c io n ro ta to ria (B uch i, R -200 ), d e ja ndo a p ro x im a d a m e n te 90 m l de un liqu ido de c o lo r oscu ro o lea g in oso . A este liqu ido se le d e n o m in o m ue s tra #23 -30.

These collected broths were combined and extracted with an equivalent volume of ethyl acetate with stirring. You will have two hours. Subsequently, the extra heat tracts will be trifuged (3 500 rpm, 30 m in u) to stop the liquid layers, according to the limitation of the action. of the layer or rg A ica for its power rp ro ce sa m ie n to. The acetyl of the type is limited to iporcomp le (<0, 3% res idu al, according to the terms of the GC / F ID) of the layer or only used Evapora tio ro ro to ria (B uch i, R -200), of 90 ml of liquid from the dark or read g in bear. This liquid is called in or more tra # 23 -30.

2. C u a n tifica c io n de F A y F A E E en la m ue s tra # 23 -30

2. C u a n tification of F A and F A E E in the month # 23 -30

U tiliza n d o un s is te m a A g ile n t 5975 B M S D eq u ipad o con una co lum n a D B -5 de 30 m x 0 ,25 m m (p e licu la de 0 ,10 |jm ), se rea lizo G C -M S . La te m p e ra tu ra de la c o lu m n a fue iso te rm ica de 3 m in u to s a 100 °C. La te m p e ra tu ra de la co lum n a se p ro g ra m o para a u m e n ta r de 100 °C a 320 °C a una ve lo c id a d de 20 °C /m in . C u an do se a lcan zo la te m p e ra tu ra fina l de 320 °C, la c o lu m n a p e rm a n e c io iso te rm ica d u ra n te 5 m in u to s a esa te m p e ra tu ra . El v o lu m e n de in ye cc io n fu e de 1 j l . E l gas po rta do r, he lio , se libe ro a 1,3 m l/m in . El e sp e c tro m e tro de m asa s e s ta b a e q u ip a d o con una fue n te de io n izac ion po r im p ac to de e lec trones . La te m p e ra tu ra de la fu e n te de io n iza c io n se es ta b le c io a 300 °C. Los pa trones de FA E E (p o r e jem p lo , d o d e ca n o a to de etilo , te tra d e c a n o a to de etilo, c is -9 -h e xa d e ce n o a to de etilo , h e xa d e ca n o a to de e tilo , o c ta d e ca n o a to de e tilo , to d o s> 99 % ); los pa tro ne s de e s te r m e tilico de ac ido graso (F A M E ) (p o r e jem p lo , d o d e ca n o a to de m etilo , te tra d e c a n o a to de m etilo , pe n ta d e ca n o a to de m etilo , cis-9-h e xa d e ce n o a to de m etilo , h e xa d e ca n o a to de m etilo , c is -11 -o c ta d e ce n o a to de m etilo , to d o s> 99 % ); tr im e tils ilil d ia z o m e ta n o (T M S D , 2 M en hexano); ac ido c lo rh id rico (37 % ); m e ta n o l (> 99 ,9 % ); y a ce ta to de e tilo (> 99 ,9 % ) se ad q u ir ie ro n en S ig m a -A ld rich y se ap lica ron sin pu rifica c ion previa .

Using a system that is 5975 BMSD eq u ipad or with a DB-5 luminaire of 30 mx 0.25 mm (eg 0.10 | jm), GC - MS. The te mpe ra tu ra of the column was iso rm ica of 3 m in u to 100 ° C. The temperature of the column will be increased to increase from 100 ° C to 320 ° C at a rate of 20 ° C / m in. When the fine temperature of 320 ° C is reached, the cooperative group is 5 meters in time for that time. The volume of in ye cc io n was 1 jl. The gas po rta do r, he lio, was released at 1.3 ml / m in. The ec eter of the three months with a single source of information from the students. The temperature of the fu in te de n iza n iza c io n is ta b le c io at 300 ° C. The patterns of FA EE (by jem p lo, where ethyl cathoa are, they are ethyl decay, c is -9-hee of ethyl ceil, he xa of ca. oc t of the type of tilo, to s>99%); The Fathers of the Fatty Acid (FAME) (Pore Jem P Lo, where I can use it as an Ethyl, I will have a Decay of Methyl, I will have an Ethyl Candidate) , cis-9- he xa of ethyl ketone, he xa of ethyl kettle, c -11-ct of ethyl kettle, to>99%); tr im e tils ilil d ia zome ta no (TMSD, 2 M in hexane); a c ido c lo rh id rico (37%); ta nol (> 99, 9%); At the end of the year (> 99.9%) it was acquired in S ig ma -A ld rich and was applied without prior notification.

La m u e s tra # 23 -30 se d e riva tizo a n a d ie n d o tr im e tils ili ld ia z o m e ta n o (T M S D ) 50 j l , H C l 8 j l y m e tan o l 36 j l a 1 m l de m ue s tra (1 m g /m l en a ce ta to de e tilo). La m ezc la se in cubo a te m p e ra tu ra am b ie n te d u ra n te 1 hora.

Sample # 23 -30 is from Riva Chavez a year ago, and it is not allowed (TMSD) 50 jl, HC l 8 jlyme tan ol 36 jla 1 ml m ue s tra (1 mg / ml in acetyl of tilo). The m ezc la was covered in te m p e ra tu ra am b ie n d u ra te te 1 hour.

A n te s de la cua n tifica c ion , se id en tifica ron los FA E E y FA M E en la m ue stra # 23 -30 u tilizan do dos m e todos . En p r im e r lugar, el t ie m p o de re te n c io n de G C de ca d a c o m p u e s to se c o m p a ro con e l t ie m p o de re ten c io n de un pa tron con oc ido . En s e g u n d o lugar, la id e n tifica c io n de cad a co m p u e s to se c o n firm o e m p a re ja n d o el e sp e c tro de m asa s del c o m p u e s to con un e sp e c tro de m asas e s ta n d a r en la b ib lio teca de esp e c tro s de m asas.

In addition to the quantification, the FA E E and FA M E were identified in method # 23 -30 using two methods. Rather, the term of retention of G C from each one with the same with the retention of a patron with a license. Secondly, the identification in each case is signed with the component of the handle of the compound with an example of masses. the b ib lio teak from esp ec tro s de t handles.

C u an do se d ispo n ia de un pa tron para un FA E E o un FA M E , la cua n tifica c ion de FA E E o FA M E se de te rm in a b a ge n e ra n d o una cu rva de ca lib ra c io n (co n ce n tra c io n fre n te a la resp ue s ta del in s tru m en to ). D espues, se ob tuvo una re lac io n linea l en tre la resp u e s ta de l in s tru m e n to y la co n ce n tra c io n de ana lito . La co n ce n tra c io n de l co m p u e s to en la m ue s tra se d e te rm in o to m a n d o su resp u e s ta de l in s tru m e n to y re fir ie n d o se a la cu rva de ca lib rac ion .

When a country is available for a FA EE or a FA ME, the qualification of FA EE or FA ME is to be issued in a cash register. ra tio n (with the fre n t ent in response to the in s tru m in to). Then, a straight line was obtained between the response of the intruder and the connection of Ana Lito. The co-ce ntra tion of the commuting in the foregoing will be sent to you in response to the response and re signed in the case of the ca lib ration.

C u a n d o un pa tron para un FA E E no es ta ba d ispo n ib le , se u tilizab a una resp ue s ta de in s tru m e n to p ro m ed io para d e te rm in a r las c o n ce n tra c io n e s de l com p ue s to . La pend ien te y la in te rse cc io n de to d a s las cu rva s de ca lib ra c io n e x is te n te s se p ro m ed ia ro n . De es to s p ro m ed ios , se d e te rm in o una re lac ion linea l en tre la c o n ce n tra c io n y la resp u e s ta de l in s tru m en to . D espues, las co n ce n tra c io n e s de co m p u e s to s d e sco n o c id o s se d e te rm in a ron re la c io n a n d o las resp ue s ta s del in s tru m en to con la re lac ion linea l en tre la resp ue s ta del in s tru m e n to y la c o n ce n tra c io n u tilizan do la Ecuacion 1.

When a sponsor for an FA EE is not ta ba d ispo n ib le, a response from an intruder is used for the purpose of demonstrating com ions of the community. The pendant and the interest of all the cava rats of the lib e r e n is te s te p ro m ed ia ro n. If this is the case, there is a line-up relationship between the control and the response of the in-service. Then, the comm nn tio ns of commutations of the sco noc ids will be given to r r in re r ing the answers of the in s tru m in t with the re lac The line between the response of the research and the con- centration used in Equation 1.

Ecuacion 1: c o n ce n tra c io n = (re sp u e s ta del in s tru m en to - in te rcep ta c ion p ro m e d io )/p e n d ie n te p ro m ed io D e sp u e s de id e n tific a r y cu a n tif ic a r el FA M A , la co n ce n tra c io n de los ac ido s g ra sos lib res a so c ia d o s se d e te rm in o en fu n c io n de la co n c e n tra c io n de F A M E y de la re lac io n de peso m o le c u la r de F A con re sp e c to a FA M E . P o r u ltim o, la c o n ce n tra c io n de F A E E y FA en m g/l se tra n s fo rm o en p o rce n ta je en la m ue s tra de b iod iese l (% p/p).

Equation 1: with ce ntra tio n = (answer of the in s tru m en to - in te rcep ta p ion p ro med io) / pend ie n te p ro m ed io D e sp u e If you want to identify the FA MA, the relationship between the two states will be free as well as two days will be available. The knowledge of FAME and the re nion of the mo r weight of the FA r with the SP ME re. First, the FAEE and FA con- centration in mg / l was carried out in the middle of the year in the middle of the year (% p / p ).

En la Tabla 8 se en um e ran las co n ce n tra c io n e s de F A E E y F A en la m u e s tra # 23 -30. La c o n ce n tra c io n to ta l de F A E E y FA fue de 80 ,7 %. El resto de los co m p u e s to s d e sco n o c id o s pueden an a liza rse po r el m e tod o LC /M S /M S . Los co m p o n e n te s p rinc ip a les de la m ue stra # 23 -30 fue ro n p e n ta d e ca n o a to de etilo, c is -9 -h e xa d e ce n o a to de etilo, h e xa d e ca n o a to de e tilo y c is -11 -o c ta d e ce n o a to de etilo.

Table 8 lists the conditions of F A E E and F A in step # 23 -30. The total cost of F A E E and FA was 80.7%. The rest of the contracts may be analyzed by the LC / M S / M S method. The com- plications of the IP address at # 23 -30 was a group of ethyl channels, that is -9-years of ethyl groups, hexes of the channels of and tilo and c is -11 -octa of ethyl acetate.

T l : P r n FAEE FA n l m r 2 -

T l: P r n FAEE FA n l m r 2 -

n in i n

n in i n

S o rp re n d e n te m e n te , la m ue s tra # 23 -30 co n te n ia F A y F A E E im pares.

S o rp re n d e n te m e n te, the most tra # 23 -30 co n te n ia F F and F A E E im pairs.

3. A n a lis is e lem e n ta l cua n tita tivo de la m u e s tra # 23 -30

3. A na lis is e lem e n ta l qua n tita tive of the m u e s tra # 23 -30

S e sa b e qu e los m e ta le s pe sad os en ven en an los c a ta liza d o re s u tilizad os en el c ra q u e o ca ta litico . P ara m e d ir los n ive le s de m e ta le s pe sad os en la m ue s tra # 23 -30 , la m u e s tra # 23 -30 se e n v io a los la b o ra to rio s G a lb ra ith , Inc., pa ra el a n a lis is cu a n tita tivo e lem e n ta l de a rsen ico , ca lc io , ca rb on o , c lo ro , coba lto , cobre, h id ro gen o , h ierro , ag ua de Karl F isher, p lom o, m an ga neso , m agnes io , m ercu rio , m o libd en o , n itrogeno, po tas io , sod io , azu fre , cinc, o x ig eno y fos fo ro . Los m e to d o s p re p a ra to r io s y a n a lit ico s se d e sc rib e n a co n tin u a c io n . Los re su lta d o s s e m u e s tra n en la Tabla 9 . T o d a s las ca n tid a d e s in d ica da s en la Tabla 9 e s tu v ie ro n por de b a jo de l n ive l de cu a n tifica c io n (LO Q , level of quantitation) e xce p to para el c a rb o n o (73 ,38 %), c lo ro (91 ppm ), h id ro g e n o (12,1 %), ag ua de Karl F ish e r (0 ,998 %), m e rcu rio (0 ,057 ppm ), o x ig e n o (14 ,53 % ) y fo s fo ro (343 ppm ). P o r lo tan to , la m ue s tra # 23 -30 no c o n te n ia n ive les a lto s de m e ta les p e sa d o s de in te res.

I know what makes me sad to see you in your classifications in the critical context. In order to go to the levels of the sadness in the month # 23 -30, the sample # 23 -30 was sent to the offices of G a lb ra ith, Inc., for the analysis is cu t tive tive and lem in arsenic, ca lc io, ca rb on o, c lo ro, coba lto, copper, id r o gen o, iron, Karl F isher, p lom o, m an ga neso, m agnes io, m ercu rio, mo libd en o, n itrogen, po tas io, sod io, azu fre, zinc, ox ig eno and fos fo ro. The lithic methods for the lithic synapses will be discussed. The re two years are shown in Table 9. In all cases in Table 9 it is your opinion under the level of the quantification (LO Q, level of quantitation) e xce p to the ca rb ono (73, 38%), c lo ro (91 ppm), h id ro geno (12.1%), ag ua de Karl F ish er (0, 998%), me rio rio (0, 057 ppm), ox ig eno (14, 53%) and fo fo fo (343 ppm). For the most part, the most recent # 23 -30 is not with them at the end of the year.

M e tod o G -52 : R e v 6: D iges tion con m ic ro o n d a s d e m u e s tra s pa ra a n a lis is d e m e ta les

METRO G -52: R e v 6: D eges tion with m icro o n d a s d e m u e s s s ra ra e n is l is d e m e ta les

En un rec ip ie n te de m ic ro o n d a s se peso una can tida d ap ro p ia d a de m u e s tra de ce rca de 0,001 g. D espues, se a n a d ie ro n los re a c tivo s a p ro p ia d o s al re c ip ie n te d e m ic ro ond as . Si se o b s e rv a b a una rea cc io n v is ib le , se d e ja b a qu e la rea cc io n ce sa ra a n te s de ta p a r el rec ip ien te . de spu es , el rec ip ie n te se s e llo y se co loco en el m ic ro o n d a s de a cu e rd o con las in s tru cc io n e s de l fa b rica n te . La te m p e ra tu ra de cad a re c ip ie n te a lca n zo un m in im o de 180± 10 °C en 5 m inu tos . Se m a n tu vo a un m in im o de 180 ± 10 °C d u ra n te 10 m inu tos. A l fina l de l p ro g ra m a de m ic ro ond as , los re c ip ie n te s se d e ja ro n e n fria r d u ra n te un m in im o de 5 m in u tos a n tes de re tira rlos . Los re c ip ie n te s se d e s ta p a ro n y se tra n s fir ie ro n a m a tra ce s v o lu m e tr ic o s pa ra su an a lis is m e d ia n te la te c n ic a ad ecu ad a .

In a wavelength vessel a weight of a sample of about 0.001 g was weighed. Subsequently, the Sap ro P ia rents are added to the repository. If a new area is observed, the area will be readable only recently. of that, the recipient was called and placed in the same wavelengths with the instructions of the manufacturer. You will have a minimum of 180 ± 10 ° C in 5 minutes. Your voice is sent at a minimum of 180 ± 10 ° C for 10 minutes. At the end of lp ro g ra ma de m ic ro ond as, the re c ip ie n te s se ja ro nen fria rdu ra n a minimum of 5 m in u cs before re throw them. Re c ip ie n te s des ta ta ro ro y se n s fir ie ro nama tra ce svo lu m e tr icos para el ani lis is med ia te tea cn ic a ad ecu ad a .

M etod o G -55 , R e v 3: B om b a de com b us tio n de o x ig eno P a rr para la d e te rm in a c io n de h a lo gen os

METHOD G -55, R e v 3: B om b a de com b us tio n o x ig eno P a rr for d e te rm in a tio n h h a lo gen os

Las m u e s tra s se p e sa ro n en una co p a de co m b u s tio n y se a n a d io ace ite m in e ra l co m o a u x ilia r de c o m b u s tio n . P ara las m e d ic io n e s de c lo ro (C I) y b ro m o (B r), se a n a d io una so lu c io n de pe ro x id o de h id ro g e n o al 1 % a la bom ba . Para las m e d ic io n e s de azu fre (S), se a n a d io una so luc io n de h id ro x id o de so d io 0,01 N. La m u e s tra y la cop a se se lla ro n en una bo m ba de co m b u s tio n de ox ig e n o P a rr ju n to con una so luc io n a b so rb e n te adecu ad a . La bo m ba se pu rgo con ox igeno , D espues, se p re su rizo a 25 -30 a tm de pres ion de ox igeno , y se encend io . A con tinu ac io n , los con ten ido s de la bo m ba se m ezc la ron b ien y se tra n s fir ie ro n a un v a s o de p re c ip ita d o s para su p o s te rio r ana lis is .

The others are entered into a co-p a co m b u s unio and added to each other in the process of communication. For the measurements of oil (CI) and white (B r), a solution of only 1% or 1% was added to the pump. ba. For the sugar measurements (S), a solution of h id ro x id o so d io 0.01 N was added. The sample and copy was called In a bus company with an ox og P p r r ju n to with a lu so nab so rb in the right way. The boom was purged with oxygen, then it was curled at 25-30 tm of oxygen pressure, and ignited. Even then, those who have had a good taste of the rum and signed a glass of water for their own potential. .

M e tod o G -30B , R e v 7: D iges tio n de cen iza h u m ed a de co m p u e s to s in o rg a n ico s y o rg an icos para an a lis is de m e ta les La m ue stra se ca rb o n izo u tilizan do H ² S O ⁴. Si se an a lizan los m e ta les que fo rm an su lfa tos inso lub les , se u tilizan H C O ⁴ y H N O ³ para ca rb o n iz a r el m a te ria l o rg an ico . D e sp u e s de c a rb o n iza r la m uestra , se an ad io H N O ³ y la m ue stra se so m e tio a re flu jo pa ra s o lu b iliz a r los m e ta le s p re sen te s . Si la so luc io n se vo lv ia tu rb ia , se a n a d ia H C l para a yu d a r a c o m p le ta r la d ige s tion . S e pu ede u sa r HF si el s ilic io e s ta ba p re se n te en la m uestra , pero so lo si el s ilic io no e ra un a n a lito de in te res. T odo el HF u tilizad o es ta ba res trin g id o a rec ip ie n tes de T e flon . El d ige rido tra n sp a re n te se tra n s fir io c u a n tita tiva m e n te a un m a traz vo lu m e tr ic o de C lase A y se llevo a un v o lu m e n fina l. D espues, la m ue s tra se ana lizo .

METHOD G -30B, R ev 7: H e c a tio n of h e m e c a c o mts to s in o rg ico only or rg an icos for me anis lis is de me ta les La m That is why they are used as H ² SO ⁴ . If the metrics that are used for their inso lub cles are analyzed, HCO ⁴ and HNO ^{3 are used} to determine the logic of the logic. In the case of the sample, HNO ^{3 was added} and the sample was included in the flow for the purposes of the study. s. If the solution was heard by you, HC l was added for a year when the decision was made. It is possible to know if the illicit is also in the sample, but only if the illicit is not an interest in interest. All of the HF is used as trin g id or recents of t e flon. The tra t sp e re r a nd re n nt was signed how much tita tive t e m a tr e m e tr ic o of C lase A and took a fine lu l l volume. Then, the sample was analyzed.

M e tod o M E -4 A R e v 2: D e te rm in a c io n de a n io n e s su p rim id o s po r c ro m a to g ra fia de iones

M e tod M E -4 A R e v 2: D e te rm in a c io n a n io n e s its p rim id o s po r c ro m a to p io trust

M e tod o S -300 R e v 7: D e te rm in a c io n de ag ua po r titu la c io n cou lo m e trica (K arl F ische r)

M e td S -300 R e v 7: D e te rm in a tio n ag ua po r titu la c io n cou m e trica (K arl F ische r)

E ste m e to d o com b in o la t itu la c io n c o u lo m e trica con la de K arl F ische r. La m u e s tra se m ezc lo con una m ezc la de a m in a -m e ta n o l que con ten ia p re d o m in a n te m e n te iones e yod o (I-) y d iox id o de azu fre . El yod o p ro du c ido en el an od o a tra ve s de la e lec tro lis is se de jo re a cc io n a r con agua. En d icho s casos, el yod o se p ro du jo en p ropo rc ion d irec ta a la can tidad de e lec tric id ad segun la Ley de Faraday. A de m as , dado que 1 m ol de agua reacc iona e s te q u io m e tr ica m e n te con ¹ m ol de yodo , ¹ m g de agua era eq u iva le n te a 10,71 cu lom b ios de e lec tric id ad . U tiliza n d o es te p rinc ip io , El m e d id o r de hu m edad d e te rm in o la ca n tid a d de ag ua d ire c ta m e n te a p a rtir de l nu m ero de cu lo m b io s n e cesa rios para la e lec tro lis is . Este p ro ced im ie n to in c luyo ta n to la in tro du cc ion d irec ta com o una te cn ica de tra ta m ie n to p re v io con vap o rizado r.

This method is com b in o la t itu la c io ncou m e trica with that of K arl F ische r. The sample was shown with a sample of am in a-me ta nol that had p re dom in an te mentions and yod o (I-) y d iox id o de azu fre. The iod op ro du c gone in the an e o r th e r th e l le tr tro lis is se j re re c a c io nar with water. In such cases, the period is directly directed at the amount of credit according to the Faraday Law. In addition, since 1 mol of reaction water is less than ¹ m iodine, ¹ mg of water was only 10.71 cu lom b ios Of electricity . Use is the main IP, the median of human age of the year in which they will depart from the number of the most important cesa rios for the e lec troph is. This process has led to the introduction of a direct treatment technique with vaping or curling r.

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continuation

M e tod o E 16-2 , rev 9 (T ra za d o E 16 -2A ): D e te rm in a c io n de azu fre u tiliza n d o el a n a liz a d o r S C -432 D R de LE C O El a n a liz a d o r de a zu fre S C -432 D R es un in s tru m e n to in fra rro jo no d ispe rs ivo , co n tro la d o d ig ita lm en te , d ise n a d o para m e d ir e l co n te n id o de a zu fre en una va rie d a d de m a te ria le s o rg a n ico s e in o rg an icos . La m ue s tra se q u em o a 1350 ± 50 °C en una a tm os fe ra de o x ig eno puro. El azu fre se ox ido a d iox id o de azu fre y se cu a n tifico po r ab so rc io n de in fra rro jo . El S C -432 D R e s ta ba e q u ip a d o con dos de tec to res , una ce lu la de in fra rro jo de a lto y ba jo a lcance .

M e tod E 16-2, rev 9 (T ra za do E 16 -2A): D e te rm in ac io n de azu frequently used by ana liz ador SC -432 DR de LE CO Ana liz a de zu fre SC -432 DR is an in s tru men to in fra rro jo no d ispe rs ivo, with the dod ig ita lm en te, designed to med go the co n te n id or from a zu fre in a variety of ways, it will be only logical. The sample is taken at 1350 ± 50 ° C in one year of pure time. The sugar was oxidized to the sugar diox and it was reported as a source of investment. The S C -432 D R e s ta ba e q u ip a d o with two tech teres, one of which is in fra rro jo de a lto and low in range.

M etod o M E -2, R e v 14: D e te rm in a c io n de ca rbono , h id ro gen o y n itrogeno

METHOD M E -2, R e v 14: D e te rm in a ca io n carbon, h id ro gen o y n itrogen

E ste in s tru m e n to q u em a la m ue stra en oxig eno puro a 950 °C en con d ic io n e s es ta ticas para p ro d u c ir p ro du c to s de co m b u s tio n de C O ² , H² O y N ². El P E -240 an a liza a u to m a tica m e n te estos p ro du c to s en un a n a liza d o r de co n d u c tiv id a d te rm ica en es tado e s ta c io n a rio de au to in teg rac ion . P ara a yu d a r a la com b ustio n , puede an a d irse a n h id r id o tu n g s tico . P ara m u e s tra s d ific iles de quem ar, puede em p le a rse un t ie m p o de co m b u s tio n p ro lo n g a d o (p o r e jem p lo , m od o de q u em a dific il).

This is the first time in pure oxygen at 950 ° C in which conditions are tactic for CO du du o co du bus co CO bus tio n de CO ² , H ² O and N ² . The EP -240 has an automatic approach to these policies in a national analyst in this state of America. ion For the purpose, you can add an identity card. In order to show the possibility of burning, a type of bus can also be used (for example, for what is difficult).

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M etod o M E -70 , R e v 4: E sp e c tro m e tria de em is ion a to m ica de p lasm a aco p la d o in d u c tiva m e n te

METHOD M E -70, R e v 4: E sp e c tro m e tr tr e m e tion of th e p icam in order to accept the m e n t

E s te m e to d o d e sc rib e d e te rm in a c io n e s de e le m e n to s m u ltip le s po r IC P -A E S u tiliza n d o s is te m a s o p tico s s im u lta n e o s y v isu a liza c io n ax ia l o rad ia l de l p lasm a. El in s tru m e n to m ide los esp e c tro s de em is ion ca ra c te ris tico s por e sp e c tro m e tria op tica . Las m ue s tra s se ne bu liza ron y el ae roso l resu ltan te se tra n sp o rto a la a n to rcha de p lasm a. Los esp e c tro s de em is ion e sp e c ifico s de e lem e n to se p ro du je ro n m ed ia n te p lasm a aco p la d o in d u c tiva m e n te a ra d io fre cu e n c ia . Los e sp ec tros se d isp e rsa ro n m ed ia n te un e sp e c tro m e tro de re jilla y las in te n s id a d e s de las line as de em is ion se co n tro la ron con d isp o s itivo s fo to se n s ib le s . P ara la de te rm in a c io n de l o ligo e lem en to , fue ne cesa rio re a liza r una co rre cc ion de fon do . El fo n d o se m id io a d yace n te a las lineas de ana lito en las m ue s tra s d u ran te el ana lis is . La pos ic ion se le cc io n a d a para la m ed ic ion de in tens idad de fondo , en uno o am b os lados de la linea an a litica , se d e te rm in o po r la com p le jid a d de l esp ec tro a d ya ce n te a la linea de ana lito . En un m odo de an a lis is , la po s ic io n u tilizada debe s e r lo m as libre pos ib le de la in te rfe ren c ia esp e c tra l y debe re fle ja r el m ism o cam b io en la in te ns idad de fo n d o que se p roduce en la long itud de onda de l an a lito m ed ida . La co rre cc ion de fo n d o no es ne ce sa ria en los caso s de e n sa n ch a m ie n to de lineas donde una m ed ic io n de co rre cc io n de fo n d o d e g ra da ria rea lm en te el resu ltad o ana litico .

This is the reason why you will be able to learn more about the IC P-AES or how to use it in the most important way. It is the radius of lp lasm a. The intru t m e n to m idem es the spe e c tro s of the ca rs c e r e c ttic r e t e r e c tro m e tria op op tica. The others have been used and the result has been translated into the last year. The e esm e es es e es e e e em e ss of e lem en to se p ro du je ro nm ed ia n te p lasm a du p in du d in tive du c a d dio fre cu en . E sp ec tros is d isp e rsa ro nm ed ia n te an e sp ec tro meter de re jilla and the inte sts of the lines of em ision rum with d isp It is important for you to know. For the purpose of carrying out the law and lem on it, it was necessary to establish a fund of connection. The model was adapted to the lines of Ana Lito in the years after the analysis. The position was given nothing for the measurement of background intensity, on one or both sides of the analytical line, the com r From the specialist, already close to the Ana Lito line. In a method of analysis, the potentialized use must be as free as possible in the international sphere and must reflect the change in the same way. In the interest of the fund that is produced in the wavelength of the long term. Funding is not necessary in the case of lines where a network of international funds is available. in the restaurant or the lithic ana.

M etod o E 80-2 , R e v 4: D e te rm in a c io n de m ercu rio (te cn ica a u to m a tiza d a de v a p o r frio )

METHOD E 80-2, R e v 4: D e te rm in a c io n de m ercu rio (technic a u to m a m a m a d a d a v a p o r cold)

E ste p ro ce d im ie n to se basa en el m e tod o E P A S W 846 7471A . La ab so rc ion a to m ica de v a p o r frio se basa en la te o ria g e ne ra l de la a b so rc ion a tom ica , que sos tie ne que los a tom os lib res de l ana lito ab so rben e n e rg ia de una fue n te de la m pa ra que es p ro po rc ion a l a la c o n ce n tra c io n de ana lito . A l u tiliza r una la m pa ra que con tiene el m eta l a m ed ir, la lo ng itud de on da exac ta ne cesa ria para la a b so rc ion se p ro du jo y las in te rfe re n c ia s se redu je ron con s id e ra b le m e n te . La a b so rc io n a to m ica de v a p o r frio u tiliza es te p rinc ip io , y los a to m o s de m ercu rio se libe ra ron al re d u c ir los iones de m ercu rio con c lo ru ro de estano (II) (S n C l)² ). El gas n itrogeno tra n sp o rta b a los a tom os a traves de una ce lu la op tica , con la la m p a ra Hg en un ex trem o y el d e te c to r en el o tro ex trem o . D eb ido a que se e m p le o el m e tod o de v a p o r frio , en lu g a r de un m e todo de llam a, los com p ue s to s o rg an icos no d ige rido s fue ro n una p re o cu p a c io n de in te rfe ren c ia , d e b ido a su am p lia band a de lo n g itu d e s de onda de abso rc ion .

This step is based on the EPASW 846 7471A method. The cold steam toxicant is based on the general theory of the acoustic blast, which has to be free of the litho abyss. There is a lamp source that is only for the purpose of Ana Lito. In light of a lamp that has the right to go, the only thing that is necessary for the abbreviation is open and the interests are needed. s were reduced with s id e ra b le ment. The use of cold steam is useful in the first place, and those from Mexico are released when the ions of the territory are released along with the course of tin (II) (S n C l) ² ). The nitrogen gas traverses the atoms through a lens of the optic, with the Hg lamp at one end and that of the te c to r at the other end. Due to the fact that the cold steam method was used, instead of a calling method, the non-governmental organizations were not allowed When he told you about his life, he had gone to his broadband of the abscess wave of his abso rtion.

Tabla 9 : Analisis elemental cuantitativo de la muestra # 23-30

Table 9: Quantitative elementary analysis of sample # 23-30

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EJEMPLO 5. PRODUCCION Y LIBERACION DE ALCOHOL GRASO DEL HOSPEDADOR DE PRODUCCION

EXAMPLE 5. PRODUCTION AND RELEASE OF ALCOHOL FAT FROM THE PRODUCTION HOST

acrl (que cod ifica la a c il-C o A red uc tasa ) se exp reso en E. coli cu ltiva d a con g lu co sa com o un ica fu e n te de ca rb o n o y ene rg ia . E. coli p rodu jo pe qu en as can tida de s de a lcoh o le s g ra sos ta le s com o d o de can o l (C ¹² : ⁰-O H ), te tra d e ca n o l (C ¹⁴ : ⁰ -O H ) y h e xa d e ca n o l (C ¹⁶ : ⁰-O H ). En o tras m uestras , FadD (a c il-C o A s in ta sa ) se exp reso ju n to con acrl en E. coli. S e ob se rvo un a u m e n to de c in co ve ce s en la p ro du cc ion de a lcoh o l graso.

acrl (which codifies the ac-C or A network uc rate) was exported in E. coli quantitative with a common interest in the capital and in the United States. E. coli p rodu jo pe que e las can tida de s de a lcoh o le sg ra sos ta le s com odo de can ol (C ¹² : ⁰ -OH), I brought you a can (C ¹⁴ : ⁰ - OH) yhe xa of can nol (C ¹⁶ : ⁰ -OH). In or after our samples, FadD (ac il-C or A s in ta sa) was reported with acrl in E. coli. A rise in volume was observed in the fatty oil sector.

En o tras m ue stra s , en un p la sm id o que lle vab a accABCD co n s tru id o com o se de sc rib e en E JE M P L O 1, acrl, fadd y accABCD (a c e til-C o A ca rb ox ilasa ), se exp resa ron ju n to con d ive rsa s t io e s te ra sa s (T E ) in d iv id u a le s en E. coli C41 (D E 3 ) de tipo s ilves tre y en una E. coli C41 (D E 3 AfadE, una cepa que ca re c ia de a c il-C o A de sh id ro g e n a sa ). Esto d io lu g a r a a u m e n to s ad ic io n a le s en la p ro d u cc io n de a lcoh o l g raso y en la m o d u la c io n de los pe rfiles de a lcoh o le s g ra so s (vea se la FIG. 6). P o r e jem p lo , la so b re e xp re s io n de 'T e sA (p E T D u e t-1 - 'T e s A ) de E. colien este s is te m a o b tu vo un au m e n to de a p ro x im a d a m e n te 60 ve ce s de C ¹² : ⁰ -O H , C ¹⁴ :⁰-O H y C m ⁰-O H , s ie nd o C ¹⁴ : ⁰ -O H el p rinc ipa l a lcoh o l graso . S e o b tuvo un resu lta d o m uy s im ila r cuando se exp reso la en z im a C h F a tB 3 (F a tB 3 de Cuphea hookeriana en p M A L -c2X -T E cu ). C u an do se exp reso la en z im a UcFatB1 (FatB1 de Umbellularia californica en pM A L -c2 X -T E u c ), la p ro du cc ion de a lcoh o l g ra so au m e n to a p ro x im a d a m e n te 20 ve ce s y C ¹² : ⁰ -O H era el a lcoho l g ra so p re do m ina n te .

On or after me, on a page that will be used as an option in the E JE MPLO 1, acrl, fadd and accABCD (ace til-C or A ca rb ox ilasa), ju r e r e r th e rsa st io es te ra sa s (TE) in d iv id ua le s in E. coli C41 (DE 3) of type s ilves tre y en an E. coli C41 (DE 3 AfadE, a strain that falls from ac il-C or A from sh id ro gena sa). This day will be added to you only in the city of Olco, and in the module of the profiles of other people (see below). FIG. 6). P o jem p lo, the so b re e xp re s io n of 'T e sA (p ETD ue t-1 -' T is A) of E. colien this s is te maob you vo an increase of ap ro x im adamen te 60 s ce C ¹² : ⁰ -OH, C ¹⁴ : ⁰ -OH and C m ⁰ -OH, s ie nd o C ¹⁴ : ⁰ -OH the beginning of the fatty lcoh ol. S eob had a domestic result and when it was exported in z im a C h F a tB 3 (F a tB 3 of Cuphea hookeriana in p MAL-c2X -TE cu). When the UcFatB1 (FatB1 of Umbellularia californica in pM AL -c2 X -TE uc) was exported, the p o du dion of a lcoh olg ra so au men t ap ro x im adamen te 20 ve ce s C ¹² : ⁰ -OH was the lcoho lg ra so p re do m ina n te.

La exp res ion de C h F a tB 3 y UcFatB1 ta m b ie n con du jo a la p ro du cc ion de ca n tid a d e s s ig n ifica tiva s de los a lcoho les g ra sos in sa tu ra d o s C ¹⁶ : ¹-O H y C ¹⁴ : ¹-O H , re sp e c tiva m e n te . T a m b ie n se en co n tra ro n a lcoh o le s g ra sos en el s o b re n a d a n te de m u e s tra s ge n e ra d a s a p a rtir de la e xp res ion de 'tesA. A 37 °C, se en co n tra ro n can tida de s a p ro x im a d a m e n te ig ua les de a lcoh o le s g ra sos en el so b re n a d a n te y en el sed im e n to ce lu la r. M ie n tras que a 25 °C, se en co n tro que a p ro x im a d a m e n te el 25 % de los a lco h o le s g ra sos e s ta ba en el so b ren ada n te . V e a se la FIG. 7.

The expiration of C h F a tB 3 and UcFatB1 ta mb ie n with du p a du r p du du cation of the a lcoho les g ra sos in sa tu ra ra two C ¹⁶ : ¹ -OH and C ¹⁴ : ¹ -OH, re sp ec tiva men te. It is also in connection with the law that you are still on the show to show yourself the main role of the testa. At 37 ° C, it is found in the presence of sap ro x im adamen that you are equal to lcoh or you are still in the so b re swim and in thirst im in ce ce r la r . After 25 ° C, it was found that 25% of the years are still acceptable in the case. See FIG. 7.

EJEMPLO 6. PRODUCCION DE ALCOHOL GRASO UTILIZANDO UNA VARIEDAD DE ACYL-COA REDUCTASAS

EXAMPLE 6. PRODUCTION OF ALCOHOL FAT USING A VARIETY OF ACYL-COA REDUCTASAS

E ste e je m p lo d e sc rib e la p ro d u cc io n de a lco h o l g ra so u tiliza n d o una va rie d a d de a c il-C o A reduc tasas . Los a lco h o le s g ra sos pueden s e r el p ro du c to fina l. C o m o a lte rna tiva , las ce lu la s ho sp e d a d o ra s de p ro du cc ion pueden g e n o m a n ip u la rse para e xp re s a r/s o b re e x p re s a r a d ic io n a lm e n te e s te r s in ta sa s para p ro d u c ir e s te re s grasos.

This is the best way to make it possible to use a series of C-C or A reduction rates. A few years ago they can be the only way to end it. As a rn tiva, the c o lu s s sp e r e p ered ra s of p ro du cc ion can genoman ip u la rse for e xp re sar / sob re exp re sarad ic io na lm en te e s te in Ta sa s para p ro duc go e s re re o fatty.

C a da uno de los cua tro genes que co d ifica b a las a c il-C o A re d u c ta sa s g ra sas (Tabla 10) de d ive rsa s fue n tes , se o p tim izo con co d o n e s para su exp re s io n en E. coli y se s in te tizo en C o don D ev ices, Inc. (C a m brid ge , M A). C a da uno de los genes s in te tiza d o s se c lono com o un fra g m e n to Ndel-AvrII en el v e c to r p C D F D u e t-1 -fa d D (de sc rito en el E je m p lo 2). C ada uno de los p la sm id o s que llevaban es to s ge ne s de a c il-C o A re d u c ta sa con el gen fadD de E. coli, se tra n s fo rm o en la cep a C41 (D E ) de E. coli (a d q u ir id a en O ve r-e xp re ss io n ).

C a gives one of the four genes that codifies the ac c-C or A du du ta sa sg ra sas (Table 10) of d ives rs s s nts, op op left with codes for their it expired in E. coli and it was reported in C o don D evices, Inc. (C am brid ge, MA). C a gives one of the genes s in te chal d two was extended as a phrase to Ndel -AvrII in the CDFD neighborhood ue t-1 -fa d D (from sc rite in E je mp lo 2) . Each one of the parents who carried them is all ac ac-C or A du du ta sa with the fadD gene of E. coli , it is translated into C41 ( DE) by E. coli (acquire id in O ve re xp re ss io n).

Las cep as re co m b in a n te s se cu ltiva ro n en 3 m l de un ca ldo LB (c o m p le m e n ta d o con e sp e c tin o m ic in a 100 m g /l) a 37 °C du ran te la noche. A 30 m l de un m ed io M 9 rec ien te (que co n te n ia e sp e c tin o m ic in a 100 m g /l) se tra n s fir ie ro n 0 ,3 m l de l cu ltivo no c tu rn o y se cu ltiva ro n a 25 °C. C u a n d o los cu ltivos a lca n za ro n una D O ⁶⁰⁰ de 0,5, se an ad io IP TG 1 m M . A cad a cu ltivo se le su m in is tro el 0,1 % de uno de los tre s ac ido s g ra sos d isu e lto s en H ² O a un pH de 7,0. Los tre s ac ido s g ra sos s u m in is tra d o s fue ro n d o d e ca n o a to de sod io , m iris ta to de sod io o pa lm ita to de sod io . C om o con tro l, ta m b ie n se inc luyo un cu ltivo sin la ad ic ion de ac ido g raso . D e sp u e s de la in du cc ion , los cu ltivos se de ja ron c re c e r a la m ism a te m p e ra tu ra d u ran te 40 ho ras m as a 25 °C.

The co mb in an te s se quantum ro n in 3 ml of a LB heat (compared to 100 mg / l) at 37 ° C du ran te the night. At 30 ml of a recent M 9 year (which includes 100 mg / l at 100 mg / l) it is signed 0, 3 ml of the quantitative number c Your rn oy is quantitative at 25 ° C. When the quantitative ones have a DO ⁶⁰⁰ of 0.5, 1 mM IP TG was added. In each case, 0.1% of one of the three courses is carried out in H ² O at a pH of 7.0. The three courses that were summed up in two years were from the Sodium, Sodium, or Sodium Pane. In all cases, a quantitative one was included without the addition of a general purpose. D e sp u e s of the in du ction, the quantitative reports are the same as the time you spend 40 hours at 25 ° C.

La cu a n tif ica c io n del re n d im ie n to de a lcoh o l g ra so al fina l de la fe rm e n ta c io n se rea lizo u tilizan do G C -M S com o se de sc rib e a n te rio rm e n te en los E JE M P L O S 3 y /o 4. En la T a b la 11 se m u e s tra el a lcoh o l g ra so re su lta n te p ro d u c id o a p a rtir de l ac ido g ra so co rre sp o n d ie n te . Los resu ltad os in d ica n que las tres a c il-C o A red uc tasa s , A cr1 , A crM y B m F A R , pu d ie ron tra n s fo rm a r los tres ac ido s g rasos en los co rre sp o n d ie n te s a lcoh o le s grasos. Los resu ltad os ta m b ie n ind ican que hF A R y J jF A R te n ia n ac tiv ida d cua nd o los su s tra to s eran m iris ta to y pa lm ita to . S in em bargo , ha b ia poca o n ing una ac tiv ida d cua nd o el sus tra to era d o de can oa to . m FAR 1 y m F A R 2 so lo de m os tra ro n baja ac tiv ida d con m iris ta to y no m ostra ro n ac tiv ida d con los o tros dos ac ido s grasos.

The typical nature of the olympic re nd im e nt e at the end of the faith in which the GC-MS was carried out or used as a guide rio rm in te in the E JE MPLOS 3 and / or 4. In T ab la 11 the a lcoh olg ra so is shown its lta n te p ro duc id oapa rtir de l ac gone g ra so co rre sp ond ie n te. The results indicate that the three ac-C or A network u s rate, A cr1, A crM and B m FAR, could be tram s fo fo rm ar the three acidic accords in the co rre sp ond ie n te sa lcoh o le s fatty. The results also indicate that hF AR and J jF AR did not take you into account when the latter were more important and pa lm ita to. However, there was little on one day of his life or his work. m FAR 1 and m FAR 2 only in the case of low income with rainbows and not less than one month with the two fatty acids.

Tabla 10: Acil-CoA reductasas

Table 10: Acyl-CoA reductases

Tabla 11: Produccion de alcohol raso

Table 11: Satin alcohol production

EJEMPLO 7. ESTERES GRASOS DE CADENA MEDIA

EXAMPLE 7. ESTERES OILS OF MEDIUM CHAIN

Las a lcoh o l ace til tra n s fe ra sa s (A A Ts, EC 2.3.1.84 ), re sp o n sa b le s de la p roducc ion de ace ta to de ac ilo en d ive rsas p lan tas, pu ed en u tiliza rse pa ra p ro d u c ir e s te re s g ra so s de lo ng itud de ca d e n a m ed ia , ta le s com o o c ta n o a to de octilo , o c ta n o a to de dec ilo , d e ca n o a to de dec ilo , y s im ila re s . Los es te re s g ra sos , s in te tiza d o s a p a rtir de a lcoh o l de cad en a m ed ia (ta l com o C6 y Ce) y la a c il-C o A o los ac ido s g ra sos de cad en a m ed ia (ta l com o C6 y Ce) tien en pu n tos de fus io n re la tiva m e n te ba jos. P o r e jem p lo , el h e xa n o a to de hex ilo tie n e un pun to de fu s io n de a p ro x im a d a m e n te -55 °C y el o c ta n o a to de oc tilo tien e un pu n to de fus io n de a p ro x im a d a m e n te - 18 ° C a a p ro x im a d a m e n te - 17 ° C. Los pu n tos de fus io n ba jos de es to s co m p u e s to s los hacen ad e cu a d o s para su uso com o b ioco m bustib le s .

The ace o ol acetil tra ns fe ra sa s (AA Ts, EC 2.3.1.84), re sp on sa b le s of the production of acetyl aceta to d ive rsas p lan tas, pu In this case, you will be able to follow it from the nudity of Canada, as it is known as Octilo, as it is from Decile, of Ca noa to de decilo, and s im ila re s. These are the following, in the case of a dosapa rtir de a lcoh ol de cad in am ed ia (ta l com o C6 and Ce) and the ac il-C or A or the ac sg ra sos de cad in am ed ia (such as C6 and Ce) they have merged in the lower tivative way. P o jem p lo, hexa noa to de hex ilo has a point of apportion at -55 ° C and the ocular position has a position of firing of ap ro x im adamen te - 18 ° C aap ro x im adamen te - 17 ° C. The melting points of these are made ad c for use with ob ioco m bustib le s.

En este e jem p lo , un gen SAAT que cod ifica una t io e s te ra sa se coe xp re so en un h o s p e d a d o r de p ro du cc ion E. coli C41 (D E 3, FadE) (com o se de sc rib e en la S o lic itud In te rn ac io na l P C T /U S 08 /058788 ) con fadD de E. coli y acrl (a lco h o l red u c ta sa de A. baylyi A D P 1 ). En el ca ld o de fe rm e n ta c io n se p ro p o rc io n o ac ido o c ta no ico . E s to d io lu g a r a la p ro du cc ion de oc ta n o a to de octilo . De m an e ra s im ila r, cua nd o el gen de e s te r s in ta sa de A. baylyi A D P 1 se e xp re so en el h o s p e d a d o r de p ro du cc ion en lu ga r de l gen SAAT, se p ro du jo oc ta n o a to de octilo .

In this example, a SAAT gene that encodes a type is known as a host of E. coli C41 (DE 3, FadE) (as of sc) It is also in the S licit In te rn ac io na l PCT / US 08/058788) with fadD of E. coli and acrl (a lco hol red uc ta sa of A. baylyi ADP 1). In the case of faith rm in ta p o p ro po rc io n or ac o oc oc no ta ico. This is the case for the oct o oc o no to to octilo. In the same way, when the genus of te inrs from A. baylyi ADP 1 is in the host of the host in place of the gen SAAT, but for Octilo .

a tra ve s de D N A 2.0 (M en lo Park, C A 94025), se s in te tizo un gen SAAT reco m b in an te . La secu en c ia de A D N s in te tiza d a se baso en la s e cu e n c ia g e n ica p u b licad a (n .° de reg is tro A F 193789 de G e n B a n k ), pero se m od ifico para e lim in a r el s itio NcoI. El gen SAAT s in te tiza d o (co m o un fra g m e n to BamHI-HindlII) se c lo no en pR S E T B (Inv itrog en , C a rlsba d , C a lifo rn ia ), linea lizado con B am H I y HindIII. El p la sm id o resu ltan te , p H Z 1.63 A se c o tra n s fo rm o en un h o s p e d a d o r de p ro du cc ion de E. coli con pA S 004.114 B , que llevab a un gen fadD de E. coli y un gen acrl de A. baylyi A D P 1. Los tra n s fo rm a n te s se cu ltiva ro n en 3 m l de un m e d io M 9 q u e c o n te n ia g lu co sa al 2 %. D e sp u e s de la in du cc ion con IP TG y de la ad ic io n de ac ido o c ta n o ico al 0 ,02 %, se de jo c re e r el cu ltivo a 25 °C du ran te 40 horas. D espues , se a n ad ie ro n 3 m l de ace ta to de ace tilo a to d o el cu ltivo y se m ezc lo va ria s ve ce s con un m ezc lad o r. La fase de a ce ta to de ace tilo se an a lizo po r G C /M S .

Through DNA 2.0 (M in Park, CA 94025), a SAAT reco mb in an te gene was found . The sequencing of DNA s in te chalda was based on the public genetic code (registration number AF 193789 of G in B ank), but it was shown to and in the NcoI site. The SAAT s in te chalk do gene (as a gmen to BamHI-HindlII) was named in pR SETB (Inv itrog en, C a rlsba d, C a lifo rn ia), line bized with B am HI and HindIII. The p la sm id o resu ltan te, p HZ 1.63 A is treated in a host of E. coli p ro du cc ion with pA S 004.114 B, which leads to a fadD gene of E. coli and an acrl gene from A. baylyi ADP 1. The measures are quantitative in 3 ml of an M 9 medium that has a 2% concentration. In the case of the introduction with IP TG and the addition of 0, 02% of ocular acid, the quantitative factor was set at 25 ° C during the 40 hours After that, 3 ml of acetyl acetyl was added to the quantitative and it was seen several times with a mixer. The acetyl acetyl phase was analyzed by GC / MS.

S o rp re n d e n te m e n te , en el e x tra c to de ace ta to de ace tilo no se ob se rvo a ce ta to de ac ilo . S in em b a rg o , se ob se rvo o c ta n o a to de octilo . S in em bargo , la cepa de con tro l sin el gen SAAT (C 41 (D E 3 , A fa d E )/p R S E T B p A S 004.114 B ) no p ro du jo o c ta n o a to de octilo . A s im ism o , la cep a (C 41 (D E 3 , A fa d E )/p H z 1.43 B p A S 004.114 B ) en la que el gen de la e s te r s in ta sa de A. baylyi A D P 1 era llevad o po r pH Z1.43 , p ro du jo o c ta n o a to de octilo (vea nse las Figs 8A-D). El ha lla zgo de que la ac tiv id a d de SAAT p ro du ce o c ta n o a to de oc tilo hace po s ib le p ro d u c ir e s te re s g ra sos de cad en a m ed ia , ta le s com o o c ta n o a to de oc tilo y d e ca n o a to de octilo , que tie n e n p u n tos de fus io n ba jos y que son ad e cu a d o s para su uso com o b io co m b u s tib le s y pa ra re e m p la za r b iod iese l ba sado en trig lice rido s .

However, in the case of acetyl oil extract, it was not referred to acyl acid. S in em ba rg o, it was obtained as octil. S in em bargo, the strain of con tro l without the SAAT gene (C 41 (DE 3, A fa d E) / p RSETB p AS 004.114 B) no p ro du jo oc ta noa to octyl. A s im ism o, the cep a (C 41 (DE 3, A fa d E) / p H z 1.43 B p AS 004.114 B) in which the gene of the te srs in ta sa of A. baylyi ADP 1 was carried by pH Z1.43, for example, octyl (see Figs 8A-D). He has said that the activity of SAAT p o du du oc oc ta noa to oc tilo has been possible for some time to go to the United States in the United States. as an ocular and of an octyl style, which have low melting points and are suitable for use as a common bussines and for the purpose of working iod iese l based on trig lice rido s.

EJEMPLO 8. PRODUCCION DE ESTERES GRASOS EN LA CEPA LS9001 DE E. C O L I

EXAMPLE 8. PRODUCTION OF FATTER ESTERS IN CEPA LS9001 OF E. COLI

A tra ve s de la g e n o m a n ip u la c io n de l h o s p e d a d o r de p ro du cc ion E. coli pa ra e xp re sa r una a c il-C o A reduc tasa , una t io e s te ra s a y una e s te r s in ta sa fo rm a d o ra de a lcoh o l graso , se p ro du je ro n es te re s g rasos. P o r tan to , el h o sp e d a d o r de p ro d u cc io n p ro du jo ta n to el lado A co m o el lado B de l e s te r y la e s tru c tu ra de am b os la do s se v io in flu e n c ia d a po r la e xp res ion de l gen de tioe s te rasa .

Through the genoman ip u the host of the p e du pion du eion E. coli pa x e r p re sa r an ac il-C or A reduction rate, a guy is saying an e s te rs in ta sa fo rm ado ra of a lcoh ol fatty, it is p ro du je ro n este te s s ras ras. P o so to, ho sp age of p ro du cc io np ro du jo ta n to side A like side B of him and r es es tru c tu ra de am b os la do s se In this way, it is possible for the uncle of the gene of uncle.

La cepa LS9001 se tra n s fo rm o con p lasm id os que llevaban un gen de e s te r s in ta sa de A. baylyi A D P 1 (p la sm id o pH Z1.43 ), un gen de t io e s te ra s a de Cuphea hookeriana (p la sm id o p M A L -c2 X -T e ch ) y un gen fadD de E. coli (p la sm id o p C D F D ue t-1 -fad ).

The LS9001 strain was carried out with p lasm id that carried a gene from this one from A. baylyi ADP 1 (p la sm id or pH Z1.43), a uncle gene is This is from Cuphea hookeriana (p la sm id op MAL -c2 X -T e ch) and a fadD gene from E. coli (p la sm id op CDFD ue t-1 -fad).

El p la sm id o pH Z 1.43 que lle vab a la e s te r s in ta sa (W S a d p l, n.° de reg is tro A A 017391 de G en B ank, EC 2.3.175 ), se co n s tru yo de la s ig u ie n te m anera . en p rim e r lugar, el gen de W s a d p l se a m p lifico con los s ig u ie n tes ceb ad o res u tilizando , com o m o lde , la s e cu e n c ia de A D N ge n o m ico de A. baylyi A D P 1 :

The p la sm id or pH Z 1.43 that arrived at this level (WS adpl, reg. No. AA 017391 of G in B ank, EC 2.3.175), is known as he left the same way. instead, the W sadpl gene was amplified with the following principles, using, as a rule, the genetic DNA signal of A. baylyi ADP 1 :

W S a d p 1 _ N d e I, 5 '-T C A T A T G C G C C C A T T A C A T C C G -3 ' (S E Q ID NO: 35); y

W S a d p 1 _ N d e I, 5 '-T C A T A T G C G C C C A T T A C A T C C G -3' (S E Q ID NO: 35); and

W S a d p 1 _ A v r, 5 '-T C C T A G G A G G G C T A A T T T A G C C C T T T A G T T -3 ' (S E Q ID N O :36).

W S a d p 1 _ A v r, 5 '-T C C T A G G A G G G C T A A T T T A G C C C T T T A G T T -3' (S E Q ID N O: 36).

D espues, el p ro du c to de la P C R se d ig irio con Ndel y A vrII y se c lo no en pC O L A D u e t-1 para d a r pH Z 1.43. D espues, el p la sm id o que llevab a wSadpl se c o tra n s fo rm o en la cepa LS 9001 de E. coli con p E T D u e t-1 'T e sA y p C D F D u e t-1 -fadD-acrl, y los tra n s fo rm a n te s se se le cc io n a ro n en p lacas LB co m p le m e n ta d a s con ka n a m ic in a 50 m g/l, c a rb e n ic ilin a 50 m g/l y e sp e c tin o m ic in a 100 mg/l.

Fter, the p ro du to c PCR d ig To call upon with NdeI and A c VRII and what not PC OLAD ue t-1 to give pH 1.43 Z. Then, the plan that led to wSadpl was co- traced in E. coli strain LS 9001 with p ETD ue t-1 'T e sA and p CDFD ue t-1 - fadD-acrl , and the works will be given to you in LB co-lacquers mentioned with ka nam ic in at 50 mg / l, ca rb en ic ilin at 50 mg / l and sp ec tin om ic in at 100 mg / l.

S e in ocu la ro n tres tra n s fo rm a n te s en 3 m l de LB K C S (ca ldo LB co m p le m e n ta d o con kan am ic ina 50 mg/l, c a rb e n ic ilin a 50 m g/l, e s p e c tin o m ic in a 100 m g/l y g lu co sa 10 g /l) y se in cub o a 37 °C en un a g ita d o r (a g ita c io n a 250 rpm ). C u an do los cu ltivos a lca n za ro n una D O ⁶⁰⁰ de a p ro x im a d a m e n te 0,5, 1,5 m l de cad a cu ltivo se tra n s fir ie ro n a m a tra ce s de 250 m l q u e co n te n ia n 50 m l/l de LB K C S . D espues , los m a tra ce s se in cu b a ro n en un a g ita d o r (250 rpm ) a 37 °C hasta que el cu ltivo a lcanzo una D O ⁶⁰⁰ de a p ro x im a d a m e n te 0 ,5 a a p ro x im a d a m e n te 1,0. D espues, se an ad io IP TG a una co n ce n tra c io n fina l de 1 m M . Los cu ltivos in du c ido s se in cub a ron a 37 °C en un a g ita d o r (250 rpm ) d u ra n te o tras 40 -48 horas.

S e in ocu la ro n three tra ns fo rm an te s in 3 ml of LB KCS (each LB com le cd with kan am ic ina 50 mg / l, ca rb en ic ilin at 50 mg / l, spec tin om ic in at 100 mg / l and lu c at 10 g / l) and was incubated at 37 ° C in an agitator (ag ata at 250 rpm). In the case of quantitative documents, an OD ⁶⁰⁰ of approximately 0.5, 1.5 ml of qualitative material will be signed for 250 ml. n n n n 50 ml / l of LB KCS. Then, the magazines are included in an agitator (250 rpm) at 37 ° C until the quantum has reached an OD ⁶⁰⁰ of ap ro x im adamen te 0.5 aap ro x im adamen te 1.0. Then, IP TG was added to a fine connection of 1 µM. In quantitative quantitative samples were incubated at 37 ° C in an agitator (250 rpm) during 40-48 hours.

Los cu ltivos se tra n s fir ie ro n de sp u e s a tu b o s con ico s de 50 m l y las ce lu la s se ce n trifu g a ro n a 3 500 X g du ran te a p ro x im a d a m e n te 10 m inu tos. C ada uno de los se d im e n to s ce lu la res se m ezc lo de spu es con 5 m l de ace ta to de etilo . Los ex trac to s de ace ta to de e tilo se an a liza ro n con G C /M S . El titu lo de es te res g ra sos (in c lu ido s C ¹⁶ C ¹⁶ , C ¹⁴ :¹ C-i⁶ , C 'i⁸ ;'iC'i⁸ ;¹ , C ² C ¹⁴ , C ² C ¹⁶ , C ² C-i⁶ :¹ , C-|⁶ C-|^6:1 and C ² C-|⁸ :¹ ) fue de a p ro x im a d a m e n te 10 m g/l. C u ando una cepa de E. coli que so lo llevab a v e c to re s v a c io s se cu ltivo en las m ism as co n d ic io n e s y s ig u ie n d o el m ism o p ro toco lo , so lo se e n co n tra ro n 0 ,2 m g/l de es te re s g ra sos en el ex trac to de a ce ta to de etilo.

The quantitative ones will be signed for your 50-meter conical period and the three times three times more than 3 500 X g. unpleasant Each of the seeds in each ce s e lc e lc of spu es with 5 ml of ethyl acetate. Extracts of ethyl acetate are analyzed with GC / MS. The title of these is (c in C s C ¹⁶ C ¹⁶ , C ¹⁴ : ¹ Ci ⁶ , C 'i ⁸ ;'iC'i⁸; ¹ , C ² C ¹⁴ , C ² C ¹⁶ , C ² Ci ⁶ : ¹ , C- | ⁶ C- | ^{6: 1} and C ² C- | ⁸ : ¹ ) was approximately 10 mg / l. When a strain of E. coli was carried, it was only carried out in the same period in the same way as the other country, I only know it. after 0.2 mg / l of these are required in the extra ethyl acetate.

EJEMPLO 9. PRODUCCION Y LIBERACION DE ESTER ETILICO GRASO DEL HOSPEDADOR DE PRODUCCION

EXAMPLE 9. PRODUCTION AND RELEASE OF ETHYLL ESTER FAT FROM THE PRODUCTION HOST

La cep a LS 9001 se tra n s fo rm o con p la sm id o s que llevab an un gen de e s te r s in ta sa de A. baylyi (p la sm id o pH Z1.43 ), un gen de t io e s te ra s a de Cuphea hookeriana (p la sm id o p M A L -c2 X -T E cu ) y un gen fadD de E. coli (p la sm id o pC D F D ue t-1 -fadD).

The LS 9001 model is carried out with p las sm id os that carried a gene from that of A. baylyi (p la sm id or pH Z1.43), a t gene It is from Cuphea hookeriana (p la sm id op MAL -c2 X -TE cu) and a fadD gene from E. coli (p la sm id or pC DFD ue t-1 -fadD).

E sta cep a re co m b in a n te se cu ltivo a 25 °C en 3 m l de un m ed io M 9 que co n te n ia ka n a m ic in a 50 m g/l, ca rb en ic ilina 100 m g/l y e sp e c tin o m ic in a 100 m g/l. D espues de la in du cc ion con IPTG , el m ed io se a jus to a una con cen tra c ion fina l de e ta no l al 1 % y g lu co sa al 2 %.

This stage was measured at 25 ° C in 3 ml of a M 9 medium that had a ka nam ic in at 50 mg / l, ca rb en ic iline 100 mg / lye sp ec tin om ic in at 100 mg / l. After the introduction with IPTG, the medium was added to one with a fine focus of 1% and 2%.

El cu ltivo se de jo c re ce r du ran te 40 ho ras de spu es de la in ducc ion con IPTG . Las ce lu la s se sep a ra ron del m ed io ag o ta d o m ed ia n te c e n trifu g a c io n a 3 500 X g d u ran te 10 m inu tos . El se d im e n to c e lu la r se resu sp e n d io con 3 m l del m ed io M9. La su sp e n s io n c e lu la r y el m e d io a g o ta d o se e x tra je ro n de spu es con 1 vo lu m e n de a ce ta to de etilo . Las fa se s de a ce ta to de e tilo resu ltan te s de la su sp e n s io n ce lu la r y el so b re n a d a n te se so m e tie ro n a an a lis is G C -M S . El e s te r e tilico C ¹⁶ fue la e sp e c ie de e s te r m as p ro m in e n te pa ra es ta tio e s te ra s a y el 20 % de l e s te r g ra so p ro d u c id o se libe ro de la ce lu la . V e a se la FIG . 9. Una cepa de con tro l C 41 (D E 3 , AfadE) de E. coli que con ten ia pC O LA D ue t-1 (v e c to r v a c io pa ra el gen de e s te r s in ta sa ), p M A L -c2 X -T E u c (que c o n te n ia fatB de U. california) y p C D F D u e t-1 -fa d D (gen FadD de E. coli), no p ro du jo ca n tid a d e s d e te c ta b le s de es te re s e tilico s de a c id o s g rasos. Los e s te re s de ac idos g ra sos se cua n tif ica ro n u tiliza n d o e s te r e tilico y ac ido p a lm itico co m e rc ia l com o re fe renc ia .

The quantitative report was given 40 hours after the IPTG injection. The news were spread from the agribusiness or the domination of the three-thousand-year-old to 3 500 Xdules for 10 minutes. The se r im in ce ce la r was resu sp end with 3 ml of the M9 medium. The first time of the year and the middle of the year were extracted with only 1 year of ethyl acetate. The factors of the ceil- ement of the tilo resu ltan te s of his sp ens io n ce lu la r and el b re nadan are being held to me by GC-MS. The latter is C ¹⁶ was the most important of these in Mexico for 20% of the years of the year. or he freed himself from the city. See FIG. 9. A strain of con C 1 C 41 (DE 3, AfadE) of E. coli that had pC O LA D ue t-1 (neighboring to the gene of this te srs in ta sa) , p MAL -c2 X -TE uc (which with the fatB of U. california) and p CDFD ue t-1 -fa d D ( FadD gene of E. coli), no p ro du jo ca n tid since te c ta b le s of this te te se se se til s of a c id osg rasos. Those of the acidic acid are known to be useful and have a political relationship with the rhetorical faith.

T a m b ie n se p re pa ra ron e s te re s g rasos u tilizan do los m e tod os de sc ritos en el p re sen te do cum e n to , excep to que se an ad io m e tan o l o iso p ro p a n o l al ca ldo de fe rm e n ta c io n . S e p ro du je ro n los e s te re s g ra sos esperados .

You also know how to follow them and use the methods of rites in the world to do so, except for what was added to me just so iso iso p ro panol al ca ldo de fe rm in ta tio n. I hope you are expecting them.

EJEMPLO 8. LA INFLUENCIA DE DIVERSAS TIOESTERASAS SOBRE LA COMPOSICION DE ESTERES ETILICOS GRASOS PRODUCIDOS EN CEPAS DE E. C O L I RECOMBINANTES.

EXAMPLE 8. THE INFLUENCE OF DIFFERENT THIOESTERASES ON THE COMPOSITION OF FATTY ETHYL ESTERS PRODUCED IN RECOMBINANT E. COLI CEPAS.

Las t io e s te ra sa s F a tB 3 (C. hookeriana) 'T e sA (E. coli) y F a tB (U. California) se exp re sa ro n s im u lta n e a m e n te con e s te r s in ta sa (de A. baylyi). S e con s tru yo un p lasm ido , pH Z1.61 , que co m p re n d ia un esq u e le to pC D FD ue t-1 (N o vag en , M ad iso n , W I) con el gen fadD, a tra v e s de l re e m p la zo de l fra g m e n to Notl-AvrlI (q u e llevab a el gen acrl) con el fra g m e n to Notl-Avrll de pH Z 1.43 de ta l m an e ra que fadD y la e s te r s in ta sa ADP1 es taban en un p lasm ido y cad a una de las se cu e n c ia s co d ifica n te s es ta ba ba jo el con tro l de un p ro m o to r deT7 d is tin to . La con s tru cc io n de pH Z1.61 h izo pos ib le u tiliza r un s is te m a de dos p lasm id os en lu g a r de l s is te m a de tre s p lasm idos . D espues, pH Z1.61 se c o tra n s fo rm o en la cep a C 41 (D E 3 , FadE) de E. coli con uno de los p lasm id os , lle va n d o ca d a uno de e llos un gen de t io e s te ra s a d ife re n te com o se de sc rib e en el p re sen te docum e n to .

The types are F a tB 3 (C. hookeriana) 'T e sA ( E. coli) and F a tB ( U. California) are expressly presented with this. rs in ta sa (from A. baylyi). It is with a plasmid, pH Z1.61, that an analysis is carried out with pD FD that was t-1 (N o vag en, M ad iso n, WI) with the gene fadD, through l re emp la zo de l fra gmen to Notl-AvrlI (which led to the acrl gene ) with the g gmen to Notl-Avrll of pH Z 1.43 of that lm an e ra that fadD y This is an ADP1 taban on a plasmid and each one of the codes is the same as the one with a single model of T7. different . The pH con- struction Z1.61 made it possible to use a two-way system instead of the last two years. Then, pH Z1.61 is measured in the form C 41 (DE 3, FadE) of E. coli with one of the plasmids, where one of them has a gene It is sad ife re n te com o se de sc rib e in the p re sen te docum en to.

Los es te res e tilico s de ac ido s g ra sos to ta le s (tan to en el s o b re n a d a n te com o en el liqu ido e tilico de ac ido s g rasos in tra ce lu la r) p ro du c ido s po r es tos tra n s fo rm a n te s se eva lu a ron u tilizan do la te cn ica de sc rita en el p resen te do cum e n to . En Tabla 12, se resum en los titu lo s y la co m p o s ic io n de los e s te re s e tilico s de ac ido s grasos.

These are the only things to be done (so in the water they swim in the liquid in the second part of the city). The only way to do this is to evaluate the use of sc rita's technical technique in the past. In Table 12, they are summarized in the titles and co m p o s ic io of the fatty acids.

Tabla 12: Titulos (mg/l) y composicion de esteres etilicos de acidos grasos mediante C41(DE3, A fa d E ) de E.

Table 12: Titles (mg / l) and composition of fatty acid ethyl esters by C41 (DE3, A fa d E) of E.

c o l i recombinante/ HZ1.61 lasmidos ue llevan dversos enes de tioesterasa.

Recombinant coli / HZ1.61 plasmids that carry reverse thioesterase enes.

EJEMPLO 9. USO DE DIVERSAS ESTER SINTASAS PARA PRODUCIR BIOCOMBUSTIBLE

EXAMPLE 9. USE OF MISCELLANEOUS ESTERS TO PRODUCE BIOFUEL

B a sa n d o se en s e cu e n c ia s de p o lin u c le o tid o s c o rre sp o n d ie n te s c o m u n ica d a s en el G e n B a n k N C B I, se s in te tiza ro n cua tro genes co d ifica n te s de e s te r s in ta sas con pe qu en as m od ifica c ion es . E stas m od ifica c io n e s inc luyen la e lim in a c io n de los s itio s de re s tr ic c io n in te rn o s Ncol, Ndel, Hindlll y A vrll s in in tro d u c ir o tros c a m b io s en la se cu e n c ia de a m in o a c id o s co rre sp o n d ie n te . C ada uno de los cua tro genes de in te res se s in te tiza ro n con un sitio Ndel en el e x tre m o 5 'y un s itio A v rll en el e x tre m o 3'. Las s e cu e n c ia s se c lo n a ro n de spu es en el s itio NdeI y A v rll de pC O LA D ue t-1 (N o va g e n e ) para p ro d u c ir p H Z 1.97 -376 , pH Z 1.97 -377 , pH Z1.97 /a tfA 1 y p H Z 1.97 /a tfA 2. En Tabla 13 s ig u ie n te , se en um e ran los p lasm id os que llevan cada uno de los cua tro genes de in te res ju n to con los re sp e c tivo s nu m ero s de reg is tro de G e n B a n k y de G e n P e p tid e .

In this regard, it is known that there is a lot of communication in the G in Bn NCBI, it is known as a number of coding genes. I know about you, with what small money it is. These measures include the limitation of the national institutions, Ncol, Ndel, Hindlll and A vrll s in tro duc ir or others. changes in the sequence of am in o a c id os cor rre sp ond ie n te. E ach one of the tro cua genes in ter est is s in te ro n chalk with an NdeI site in the former tre mo 5 'and ite s A v rll in ex tre mo 3'. The signs are reported only in the NdeI and A v rll of pC O LA D ue t-1 (N o va gene) for p du du go p HZ 1.97-376, pH Z 1.97 -377, pH Z1.97 / a tfA 1 and p HZ 1.97 / a tfA 2. In Table 13 s also, the plasmids carrying each of the four are shown. genes of interest with the representative numbers of registers of G in B anky of G in P ep t e.

Tabla 13: Ester sintasas

Table 13: Ester synthases

Cada uno de los cuatro plasmidos se transformo en la cepa C41 (DE3, AfadEAfabR)/pETDuet-1-'TesA + pCDFDuet-1-fadD de E. coli. Se seleccionaron tres transformantes de cada transformacion para estudios de fermentacion para determinar sus habilidades para sintetizar esteres etilicos de acidos grasos. La etapa de fermentacion se realizo como se describe en el EJEMPLO 6, pero a dos temperaturas diferentes, 25 °C o 37 °C. La cepa C41 (DE3, AfadEAfabR)/pETDuet-1-’TesA + pCDFDuet-1-fadD+pHZ1.43 (que expresa la ADP1 ester sintasa) se utilizo como control positivo y la cepa C41 (De 3, AfadEAfabR)/pETDuet-1-'TesA pCDFDuet-1 -fadD como control negativo.

Each of the four plasmids was transformed into strain C41 (DE3, AfadEAfabR) / pETDuet-1-'TesA + pCDFDuet- 1-fadD from E. coli. Three transformants of each transformation were selected for fermentation studies to determine their abilities to synthesize fatty acid ethyl esters. The fermentation stage was performed as described in EXAMPLE 6, but at two different temperatures, 25 ° C or 37 ° C. The strain C41 (DE3, AfadEAfabR) / pETDuet-1-'TesA + pCDFDuet-1-fadD + pHZ1.43 (ADP1 expressing ester synthase) was used as positive control and strain C41 (From 3 AfadEAfabR) / pETDuet -1-'TesA pCDFDuet-1 -fadD as a negative control.

La expresion de cada uno de los cuatro genes de ester sintasa en la cepa de E. coli con actividad fadE y fabRy atenuada y sobreexpresion de 'tesA y fadD, permitio que cada cepa produjera aproximadamente 250 mg/l de FAe E a 25 °C. Esta fue la misma cantidad producida por el control positivo que expresaba la ADP1 ester sintasa. En cambio, la cepa de control negativo produjo menos de 50 mg/l de FAEE en las mismas condiciones a 25 °C (vease, la FIG. 10). La composicion de acilo graso de FAEE producida a partir de estas cuatro ester sintasas fue similar a la de las ADP1 ester sintasas (vease, la FIG. 11).

The expression of each of the four ester synthase genes in the E. coli strain with attenuated fadE and fabRy activity and overexpression of 'tesA and fadD, allowed each strain to produce approximately 250 mg / l FAe E at 25 ° C . This was the same amount produced by the positive control expressed by the ADP1 ester synthase. In contrast, the negative control strain produced less than 50 mg / l of FAEE under the same conditions at 25 ° C (see, FIG. 10). The fatty acyl composition of FAEE produced from these four ester synthases was similar to that of the ADP1 ester synthases (see, FIG. 11).

Los resultados de las fermentaciones realizadas a 37 °C indicaron que las cepas que llevaban pHZ1.97_aftA2 y las que llevaban pHZ1.97_376 produjeron mas FAEE que el control positivo que llevaba pHZ1.43 (vease, la FIG. 12). Las cepas que llevaban pHZ1.97_aftA2 y las que llevaban pHZ1.97_376 tambien produjeron una gran cantidad de acido graso libre (vease, la FIG. 13). Mientras que la cepa que llevaba pHZ.143 no acumulo acido graso libre. Los resultados demostraron que estas cuatro ester sintasas eran capaces de aceptar etanol y una amplia gama de acil-CoA como sustratos.

The results of the fermentations carried out at 37 ° C indicated that the strains carrying pHZ1.97_aftA2 and those carrying pHZ1.97_376 produced more FAEE than the positive control carrying pHZ1.43 (see, FIG. 12). Strains that carried pHZ1.97_aftA2 and those that carried pHZ1.97_376 also produced a large amount of free fatty acid (see, FIG. 13). While the strain that carried pHZ.143 did not accumulate free fatty acid. The results showed that these four ester synthases were able to accept ethanol and a wide range of acyl-CoA as substrates.

EJEMPLO 12. USO DE ESTER SINTASA EUCARIOTA PARA PRODUCIR BIOCOMBUSTIBLE

EXAMPLE 12. USE OF EUCARIOTE SINTASA ESTER TO PRODUCE BIOFUEL

Este ejemplo describe la clonacion y expresion de una ester sintasa de Saccharomyces cerevisiae. Los plasmidos se generaron utilizando tecnicas de biologia molecular convencionales.

This example describes the cloning and expression of an ester synthase of Saccharomyces cerevisiae. Plasmids were generated using conventional molecular biology techniques.

Tabla 14: Plasmidos con e e b l

Table 14: Plasmids with eebl

Para expresar los diversos plasmidos, se utilizo un hospedador de produccion C41 (DE3 AfadE) de E. coli. Se cultivaron celulas de E. coli en un medio minimo M9 (que contenia 6 g/l de Na² HPO⁴ , 3 g/l de KH² PO⁴ , 0,5 g/l de NaCl, 1 g/l de NH⁴ Cl, 1 mg/l de tiamina (vit. B1), MgSO⁴ 1 mM, CaCh 0,1 mM, glucosa al 0,4 % (p/v) o al 2 % (p/v)). Todas las soluciones madre de acidos grasos se prepararon disolviendo la sal sodica o potasica del acido graso en agua destilada desinoizada a pH 7,0. La solucion madre de acido octanoico se adquirio en Sigma, St. Louis, MO.

To express the various plasmids, a host production C41 (DE3 AFAD) E. coli was used. E. coli cells were cultured in a minimum M9 medium (containing 6 g / l Na ² HPO ⁴ , 3 g / l KH ² PO ⁴ , 0.5 g / l NaCl, 1 g / l NH ⁴ Cl, 1 mg / l thiamine (vit. B1), 1 mM MgSO ⁴ , 0.1 mM CaCh, 0.4% (w / v) or 2% (w / v) glucose). All stock solutions of fatty acids were prepared by dissolving the sodium or potassium salt of the fatty acid in distilled distilled water at pH 7.0. The stock solution of octanoic acid was purchased from Sigma, St. Louis, MO.

Las fermentaciones se realizaron utilizando la cepa C41 (DE3 AfadE) que contenia los plasmidos pCDFDuet-1-acr1, pMAL-c2X-TEuc (ucFatB) y pGL10.59 (eebl). La cepa control fue C41 (DE3 AfadE) que llevaba pCDFDuet-1-acr1, pMAL-c2X-TEuc, y el vector vacio pCOLADuet-1. Cada una de las tres colonias de cada transformacion se utilizo para inocular un cultivo iniciador de M9 glucosa al 0,4% complementado con carbenicilina (100|jg/ml), espectinomicina (100 jg/m l) y kanamicina (50 jg/ml). Los cultivos se dejaron crecer a 37 °C durante la noche. Los cultivos de produccion se establecieron produciendo una dilucion 1: 100 de cultivo iniciador para inocular 3 ml de medio M9 glucosa al 0,4 %. Los cultivos de produccion se dejaron crecer a 37 °C hasta alcanzar una DO⁶⁰⁰ = 0,6 antes de inducirse con IPTG 1 mM, se suministro etanol al 1 % y se cultivo durante 40 horas mas a 25 °C. Los cultivos celulares completos se extrajeron con un mismo volumen de acetato de etilo mediante agitacion en vortex (con una agitadora vorticial) durante 30 segundos. La fase organica se tomo y se examino en el GC/MS utilizando el metodo alkane_1_splitless_ctc.m para la deteccion de FAEE, que se describe anteriormente en el EJEMPLO 4, parte 2, "Cuantificacion de FA y FAEE en la muestra # 23-30".

Fermentations were performed using strain C41 (DE3 AfadE) containing plasmids pCDFDuet-1-acr1, pMAL-c2X-TEuc ( ucFatB) and pGL10.59 ( eebl). The control strain was C41 (DE3 AfadE) carrying pCDFDuet-1-acr1, pMAL-c2X-TEuc, and the empty vector pCOLADuet-1. Each of the three colonies of each transformation was used to inoculate a 0.4% M9 glucose starter culture supplemented with carbenicillin (100 | jg / ml), spectinomycin (100 jg / ml) and kanamycin (50 jg / ml) . The cultures were allowed to grow at 37 ° C overnight. Production cultures were established producing a 1: 100 dilution of starter culture to inoculate 3 ml of 0.4% M9 glucose medium. The production cultures were allowed to grow at 37 ° C until an OD ⁶⁰⁰ = 0.6 was reached before inducing with 1 mM IPTG, 1% ethanol was supplied and cultivated for an additional 40 hours at 25 ° C. Complete cell cultures were extracted with the same volume of ethyl acetate by vortexing (with a vortex shaker) for 30 seconds. The organic phase was taken and examined in the GC / MS using the alkane_1_splitless_ctc.m method for the detection of FAEE, which is described above in EXAMPLE 4, part 2, "Quantification of FA and FAEE in sample # 23-30 ".

No se detectaron picos de FAEE en las muestras. Para determinar si eebl se expresaba correctamente, los cultivos inducidos con IPTg y los no inducidos con IPTG, se analizaron por SDS-PAGe . No se detecto ninguna banda correspondiente al tamano de Eeb1 (de aproximadamente 52 kDa). Esto sugirio que, en este sistema de plasmido particular, Eeb1 no se expresa bien.

No FAEE peaks were detected in the samples. To determine if eebl was expressed correctly, cultures induced with IPTg and those not induced with IPTG were analyzed by SDS-PAGe. No band corresponding to the size of Eeb1 (approximately 52 kDa) was detected. This suggested that, in this particular plasmid system, Eeb1 is not well expressed.

Se realizaron experimentos de expresion adicionales utilizando un vector de expresion diferente. El gen se clono en el vector pMALc2x, que expresaba la proteina diana como una fusion a proteinas de union a maltosa (MBP, del ingles maltose binding protein). El analisis por SDS-PAGE de los lisados de celulas completos revelo que los cultivos inducidos con IPTG 1 mM produjeron una banda de tamano apropiado correspondiente a la fusion Eeb1-MBP (de aproximadamente 92 kDa). En las celulas no inducidas la banda no estaba presente. Este experimento se describio en detalle en la Solicitud Internacional No. PCT/US08/058788.

Additional expression experiments were performed using a different expression vector. The gene was cloned into the pMALc2x vector, which expressed the target protein as a fusion to maltose binding proteins (MBP ). SDS-PAGE analysis of whole cell lysates revealed that cultures induced with 1 mM IPTG produced a band of appropriate size corresponding to the Eeb1-MBP fusion (approximately 92 kDa). In non-induced cells the band was not present. This experiment was described in detail in International Application No. PCT / US08 / 058788.

La actividad enzimatica de Eeb1 se evaluo utilizando la cepa C41 (DE3 AfadE) de E. coli que llevaba los plasmidos pCDFDuet-1-acr1 y pGL10.104 (eeb1). Una cepa C41 (d E3 AfadE) con pCDFDuet-1-acr1 y pMALc2x sirvio como cepa de control. Se seleccionaron tres colonias de cada transformacion y cada una de ellas se utilizo para inocular un cultivo iniciador durante la noche de medio M9 glucosa al 0,4 % complementado con carbenicilina (100 |jg/ml) y espectinomicina (100|jg/ml). Se utilizo una dilucion 1: 100 del cultivo iniciador para inocular 10 ml de cultivos de produccion en medio M9 glucosa al 0,4 %. Los cultivos de produccion se dejaron crecer a 37 °C hasta alcanzar una DO⁶⁰⁰ = 0,4 - 0,5 antes de la induccion con IPTG 1 mM. A cada uno de los cultivos se les suministro aproximadamente etanol al 1 %, acido octanoico (a aproximadamente 0,01 % o a aproximadamente 0,02 % del volumen final) y/o acido decanoico (a aproximadamente 0,02 % del volumen final). Las fermentaciones prosiguieron durante 24 horas a 25 °C. Las extracciones se llevaron a cabo anadiendo al cultivo un volumen 1/10 de HCl 12 M y un mismo volumen de acetato de etilo y agitando en vortex durante 30 segundos. Las muestras se analizaron mediante GC/MS como se describe anteriormente.

The enzymatic activity of Eeb1 was evaluated using E. coli strain C41 (DE3 AfadE) carrying plasmids pCDFDuet-1-acr1 and pGL10.104 (eeb1). A C41 strain (d E3 AfadE) with pCDFDuet-1-acr1 and pMALc2x served as a control strain. Three colonies of each transformation were selected and each of them was used to inoculate an overnight culture of 0.4% M9 glucose medium supplemented with carbenicillin (100 | jg / ml) and spectinomycin (100 | jg / ml) . A 1: 100 dilution of the starter culture was used to inoculate 10 ml of production cultures in 0.4% M9 glucose medium. Production cultures were grown at 37 ° C until OD ⁶⁰⁰ = 0.4-0.5 before induction with 1 mM IPTG. Each of the cultures were supplied with approximately 1% ethanol, octanoic acid (approximately 0.01% or approximately 0.02% of the final volume) and / or decanoic acid (approximately 0.02% of the final volume). . The fermentations continued for 24 hours at 25 ° C. The extractions were carried out by adding to the culture a 1/10 volume of 12M HCl and the same volume of ethyl acetate and vortexing for 30 seconds. Samples were analyzed by GC / MS as described above.

Los datos de GC/MS revelaron que puede detectarse un pico correspondiente al ester etilico del acido octanoico para las celulas que expresan eebl y suministrando acido octanoico y etanol. La cepa de control de vector tambien mostro un pico de C² C⁸ , aunque sea un pico mas pequeno que el de las celulas que expresan eeb1.

Data GC / MS revealed to be the ethyl ester of octanoic acid for expressing cells and supplying eebl octanoic acid and ethanol a peak corresponding detected. The vector control strain also showed a peak of C ² C ⁸ , although it is a smaller peak than that of cells expressing eeb1.

Las celulas a las que se suministro acido decanoico al 0,02 % no crecieron bien; por lo tanto, se realizaron los siguientes estudios utilizando acido decanoico al 0,01 % o al 0,005 %. Para probar la capacidad de Eeb1 para utilizar alcoholes distintos al etanol en la sintesis de esteres de acidos grasos, se realizaron fermentaciones utilizando la misma cepa: C41 (DE3 AfadE) con pCDFDuet-1-acr1 y pGL10.104. Las celulas se cultivaron como se describio anteriormente. En la induccion, a las celulas se las suministro acido octanoico al 0,02 % junto con metanol, etanol, propanol o isopropanol al 1 %. A las celulas tambien se las suministro acido decanoico al 0,01 % o 0,005 % y etanol al 1 %. Se permitio que las fermentaciones continuaran despues de la induccion durante 24 horas a 25 °C. Para preparar el analisis por GC/MS, los cultivos se centrifugaron para separar el sedimento y el sobrenadante. El sedimento se resuspendio en un mismo volumen de medio M9 glucosa al 0,4% reciente. Tanto los sedimentos resuspendidos como las muestras de sobrenadante se extrajeron como se describe anteriormente y se analizaron mediante GC/MS.

The cells to which 0.02% decanoic acid was supplied did not grow well; therefore, the following studies were performed using 0.01% or 0.005% decanoic acid. To test the ability of Eeb1 to use alcohols other than ethanol in the synthesis of fatty acid esters, fermentations were performed using the same strain: C41 (DE3 AfadE) with pCDFDuet-1-acr1 and pGL10.104. The cells were cultured as described above. On induction, the cells were supplied with 0.02% octanoic acid together with methanol, ethanol, propanol or 1% isopropanol. The cells were also supplied with 0.01% or 0.005% decanoic acid and 1% ethanol. The fermentations were allowed to continue after induction for 24 hours at 25 ° C. To prepare the GC / MS analysis, the cultures were centrifuged to separate the sediment and the supernatant. The sediment was resuspended in the same volume of fresh 0.4% M9 glucose medium. Both resuspended sediments and supernatant samples were extracted as described above and analyzed by GC / MS.

Todas las muestras de sobrenadante contenian grandes cantidades de acido graso pero no esteres de acido graso detectables. De manera similar, las muestras de sedimento de control de vector no contenian picos de ester de acido graso, segun lo determinado utilizando GC/MS. Sin embargo, celulas a las que se suministro acido graso C¹⁰ , mostraron picos que se identificaron como representativos de acido decanoico.

All supernatant samples contained large amounts of fatty acid but no detectable fatty acid esters. Similarly, the vector control sediment samples did not contain fatty acid ester peaks, as determined using GC / MS. However, cells to which C ¹⁰ fatty acid was supplied showed peaks that were identified as representative of decanoic acid.

Las muestras de sedimento procedentes de las celulas que expresan Eeb1 y a las que se suministro acido graso C8 y propanol o etanol, mostraron picos pequenos correspondientes a esteres de propilo o etilo. En las celulas a las que se suministro metanol o isopropanol, no se detecto ningun pico. Los cultivos a los que se suministro un acido graso C¹⁰ y etanol al 0,01 % o al 0,005 %, tambien produjeron FAEE C² C¹⁰ , pero se encontro FAEE en las muestras de sedimento.

The sediment samples from the cells expressing Eeb1 and to which C8 fatty acid and propanol or ethanol were supplied, showed small peaks corresponding to propyl or ethyl esters. In the cells to which methanol or isopropanol was supplied, no peak was detected. Cultures that were supplied with a C ¹⁰ fatty acid and 0.01% or 0.005% ethanol also produced FAEE C ² C ¹⁰ , but FAEE was found in sediment samples.

Los resultados indicaron que Eeb1 era capaz de sintetizar FAEE utilizando acidos octanoico o decanoico, y que tambien podia utilizar metanol para generar el ester metilico octanoico. Sin embargo, estos compuestos eran muy volatiles y, como tales, los datos de GC/MS podrian no haber reflejado con precision los verdaderos titulos. Para medir con mayor precision la formacion de productos, se utilizo una capa de hexadecano para facilitar la captura de FAEE mas volatiles.

The results indicated that Eeb1 was able to synthesize FAEE using octanoic or decanoic acids, and that it could also use methanol to generate the octanoic methyl ester. However, these compounds were very volatile and, as such, the GC / MS data may not have accurately reflected the true titles. To more accurately measure product formation, a hexadecane layer was used to facilitate the capture of more volatile FAEE.

La actividad de Eeb1 con respecto a los sustratos de acidos grasos se evaluo utilizando la cepa C41 (DE3 AfadE) con pCDFDuet-1-acr1 y pGL10.104, a la que se suministro acidos grasos de diferentes longitudes de cadena. Las celulas se cultivaron como se describe anteriormente, pero para promover un mejor crecimiento celular despues de la induccion, se indujeron a una DO600=0,8-0,9. En este punto, a las celulas se las suministro etanol al 1 % y un acido graso C8 al 0,02 % o una combinacion al 0,01 % de los siguientes acidos grasos: C¹⁰ , C¹² , C¹⁴ y C¹⁶. A los cultivos a los que se les suministro acidos grasos C8 o C¹⁰ , se cubrieron con un volumen total de hexadecano al 20 %. Las fermentaciones se llevaron a cabo durante 24 horas mas a 25 °C despues de la induccion. Para el analisis de los productos, se extrajeron cultivos completos (sin separar el sobrenadante del sedimento) como se describe en el presente documento, con un volumen 1/10 de HCl y un mismo volumen (al volumen de cultivo) de acetato de etilo. Las muestras tratadas con hexadecano se inyectaron directamente en el GC/MS utilizando el programa hex_1_splitless_ctc.m, que se describe anteriormente en el EJEMPLO 4, parte 2, "Cuantificacion de FA y FAEE en la muestra # 23-30".

Eeb1 activity with respect to fatty acid substrates was evaluated using strain C41 (DE3 AfadE) with pCDFDuet-1-acr1 and pGL10.104, to which fatty acids of different chain lengths were supplied. Cells were cultured as described above, but to promote better cell growth after induction, they were induced at OD600 = 0.8-0.9. At this point, the cells were supplied with 1% ethanol and a 0.02% C8 fatty acid or a 0.01% combination of the following fatty acids: C ¹⁰ , C ¹² , C ¹⁴ and C ¹⁶ . The cultures that were supplied with C8 or C ¹⁰ fatty acids were covered with a total volume of 20% hexadecane. The fermentations were carried out for an additional 24 hours at 25 ° C after induction. For the analysis of the products, whole cultures (without separating the supernatant from the sediment) were extracted as described herein, with a 1/10 volume of HCl and the same volume (to the culture volume) of ethyl acetate. Samples treated with hexadecane were injected directly into the GC / MS using the hex_1_splitless_ctc.m program, which is described earlier in EXAMPLE 4, part 2, "Quantification of FA and FAEE in sample # 23-30".

Ninguno de los controles de vector tenia picos de FAEE detectables. Para las celulas a las que se suministro C8 y C¹⁰ , en las muestras de hexadecano, se detectaron picos grandes de C² C⁸ y de C² C¹⁰ , pero no en las muestras de acetato de etilo. Esto demostro que el hexadecano fue capaz de atrapar satisfactoriamente los FAEE volatiles. Para el resto de las m ue s tra s de a ce ta to de etilo, se d e tec ta ron p icos pe qu en os para los FA E E C ² C ¹² y C ² C ¹⁴ , pero no pa ra F A E E C ² C ¹⁶. P o r tan to , Eeb1 g e n e ra e s te re s e tilico s u tiliza n d o a c id o s g ra sos con lo n g itu d e s de ca d e n a de C8 a C ¹⁴. Eeb1 fa vo re c io a los ac ido s g ra sos C ⁸ y C ¹⁰ sob re los ac ido s g ra sos de cad en a m as larga.

None of the vector controls had detectable FAEE peaks. For the cells to which C8 and C ¹⁰ were supplied, in the hexadecane samples, large peaks of C ² C ⁸ and C ² C ¹⁰ were detected, but not in the ethyl acetate samples. This showed that hexadecane was able to successfully trap volatile FAEE. For the rest of the years of ethyl acetate, small peaks were developed for FA EEC ² C ¹² and C ² C ¹⁴ , but not for FAEEC ² C ¹⁶ . For that reason, Eeb1 generates the use of his credit with the channel numbers from C8 to C ¹⁴ . Eeb1 fa vo re c io the ac se cse s s ra ra are C ⁸ and C ¹⁰ sb ra s s s ra ra are de cad in longer.

EJEMPLO 13. INTEGRACION GENOMICA DE SECUENCIAS RECOMBINANTES PARA FABRICAR UNA CEPA HOSPEDADORA QUE SOBREEXPRESE LOS GENES F A B A Y/O F A B B DE E. C O LI.

EXAMPLE 13. GENOMIC INTEGRATION OF RECOMBINANT SEQUENCES TO MANUFACTURE A HOSPEDING CEPA THAT OVEREXPRESSES THE FABA AND / OR FABB GENES OF E. CO LI.

S e sabe que el p ro d u c to de l gen fabR ac tu a co m o un re p re so r de la e xp res ion de los ge ne s faba y fabB. T a m b ie n se sabe que F adR fu n c io n a com o un a c tiv a d o r de los m ism os genes. El FabR y las se cu e n c ia s de un ion con sen so e sp e ra d a s fu e ro n p u b lica d a s p re v ia m e n te po r Z h a n g e t al., J. B iol. Chem. 277: 15558 -15565 , 2002. En la FIG. 14 se m ue s tra n las s e cu e n c ia s de un ion co n se n so y sus ub ica c io n e s re la tiva s a los ge ne s faba y fabB de E. coli.

It is known that the p du du of the gene fabR ac you as a re p re so r of the e xp res ion of the ge ne s faba and fabB. It is also known that F adR was only a common agent of the same genes. The FabR and the members of an ion with a sense of interest were the public policy of Z hanget al., J. Bol. Chem. 277: 15558-1565, 2002. In FIG. 14 is behind the signs of a common union and its location is the main faba and fabB of E. coli.

S e creo una cepa fabR in ac tiva da de E. coli. P a ra a m p lif ic a r a p ro x im a d a m e n te 1 000 pb de fabR en d irecc io n 5' se u tiliza ron los ceb ad o res T rm A _ R _ N o tI y F abR _F O P y para a m p lif ic a r a p ro x im a d a m e n te 1 000 pb de fabR en d ire cc io n 3' se u tiliza ro n los c e b a d o re s S th A _ F _ B a m y F a b R _ R O P . P ara c re a r una c o n s tru cc io n para la de le c io n en m arco del gen fabR completo, se a p lico PC R con sup e rp os ic ion . La con s tru cc io n pa ra la de le c ion de fabR se c lono en un p lasm id o pK O V 3 sen s ib le a tem p e ra tu ra , que con ten ia S acB para la se lecc io n po r con tras te . S e rea lizo una de le c ion c ro m o so m ica de fabR segun el m e tod o de sc rito en L ink et al., J. B act., 179 :6228 -6237 , 1997.

S e created a fabR in ac tiva da E. coli strain . P a ra amp lif ic arap ro x im adamen te 1000 bp of fabR in 5 'direction the primers are used T rm A _ R _ N or tI and F abR _F OP and for amp Lif ic arap ro x im adamen te 1 000 bp of fabR in the direction 3 'is used in the priming re s S th A _ F _ B am and F ab R _ ROP. In order to build a structure for reading in the arc of the complete manufacturing gene , PC R was applied with the assumption of the condition. The answer to the work of fabR was extended in a pasm id or pK OV 3 only if you were afraid of it, which had S acB for reading. n po r with after you. One of the most important works of the art was made according to the method of science in L ink et al., J. B act., 179: 6228-6237, 1997.

Tabla 15: cebadores de fa b R in a c t iv a d o s

Table 15: primers of fa b R in act iv ados

EJEMPLO 14. CONSTRUCCION DE HOSPEDADORES DE PRODUCCION

EXAMPLE 14. CONSTRUCTION OF PRODUCTION HOSPEDADORES

La Tabla 16 id e n tifica los ho m o lo g o s de c ie rto s genes d e sc rito s en el p re sen te d o cu m e n to , que se sabe que se exp resa n en m ic ro o rg a n ism o s que p roducen b iod iese l, a lco h o le s g ra sos e h id ro ca rbu ros . Para a u m e n ta r la p ro du cc ion de ac ido s g ra sos y, po r lo tan to , la p ro du cc ion de h id ro ca rb u ro s en h o sp e d a d o re s de p ro du cc ion , ta les com o los id e n tifica d o s en la Tabla 16, pueden e xp re sa rse en ge ne s he te ro logo s , ta le s com o los de E. coli.

Table 16 identifies the genes of the sc rite s genes in the p r e rient, which is known to be expressed in micro or rg an isms. that produce b iod iese l, a lco ho le sg ra sos eh id ro ca rbu ros. To increase the coverage of the accentuated system and, therefore, the extent of the identity of the state in which it has been established in the future Ion, as they are listed in Table 16, can be found in general terms, such as those of E. coli.

Un e xp e rto h a b itu a l en la m a te ria a p re c ia ra que u tiliza n d o los m e to d o s d e sc rito s en el p re se n te do cu m e n to , ta m b ie n pu ed en exp re sa rse , so b re e x p re s a rs e o a te n u a rse ge ne s que son e n d o g e n o s a los m ic ro o rg a n ism o s p ro p o rc io n a d o s en laTabla 16. A de m as , los genes que se d e sc rib e n en la Tabla 16 pueden exp resa rse , s o b re e xp re sa rse o a te n u a rse en ho sp e d a d o re s de p ro du cc ion que p roducen h id ro ca rb u ro s de m an e ra e n d o ge na para p e rm itir la p ro du cc ion de h id ro ca rb u ro s e sp e c ifico s con lo ng itud de cadena de ca rbono , pu n tos de sa tu ra c io n y pu n tos de ra m ifica c io n de fin idos.

An e xp e rto hab itu al in the ma ap ria ap re c ia ra that uses the methods of the rite s in the p r se n t do te men t, as well as pu pu in exp re sa rse, so b re exp re sa rs eoa te nua rse ge ne s that the m icro or rg an ism osp ro po rc io swim in the Table 16. In addition, the genes that are They can be explored in Table 16 on the basis of which they have been carried out in the current period of time that they produce in the same way. For the end of the year, it is the same as the chain of carbon chain, as well as your network. ratios of finite acids.

co n tin u a c io n

continuation

co n tin u a c io n

continuation

co n tin u a c io n

continuation

co n tin u a c io n

continuation

co n tin u a c io n

continuation

Los nu m ero s de reg is tro de la T a b la 16 son de l G e n B ank , V e rs io n 159.0 de l 15 de ab ril de 2007 , los n u m e ro s EC de la T a b la 16 son de la K E G G , V e rs io n 42.0 de ab ril de 2007 (m a s a c tu a liza c io n e s d ia ria s hasta el 9 de m ayo de 2007 in c lus ive ), los resu ltad os de la cepa E a273 de Erwinia amylovora se ob tu v ie ro n en el ce n tro de se cu e n c ia c io n de S anger, la se cu e n c ia a le a to ria se co m p le to el 9 de m ayo de 2007 , las po s ic io ne s de Erwinia rep rese n tan ub ica c ion es en el p s e u d o c ro m o so m a de S ange r, las se cu e n c ia s de Vibrio furnisii M1 son de l p s e u d o c ro m o so m a V FM 1, co n s tru cc io n v2, del 28 de se p tie m b re de 2006, e in c luye el gen com p le to , y ta m b ie n puede in c lu ir la secu en c ia flan qu ea n te .

The registration numbers of T ab la 16 are from l G in B ank, V e rs io n 159.0 from l April 15, 2007, the EC numbers of T ab la 16 are from the KEGG, V e rs io n 42.0 of April 2007 (more than until the end of the year until 9 of 2007 in May 2007), the results of the strain E a273 of Erwinia amylovora will see you in the heart of the anger department, the court will tell you how it was taken on the 9th of May As of 2007, Erwinia's poetry is so ubiquitous in the pseudoc ro mo so ma of S ange r, those of Vibrio furnisii M1 are from lpseudoc ro mo so ma V FM 1, co ns tru cc io n v2, dated September 28, 2006, and in which the gene is shared, and also the section can go to the flan what's up

EJEMPLO 15. CEPAS DE PRODUCCION EJEMPLARES ADICIONALES

EXAMPLE 15. PRODUCTION CEPAS ADDITIONAL EXAMPLES

La Tabla 17 p ro p o rc io n a cep as de p ro du cc ion e jem p la re s ad ic io na le s . S e de sc rib en dos e je m p lo s de ru tas b io s in te tica s para la p ro du cc ion de ac ido s grasos, a lcoh o le s g ra sos y es te res de cera . P o r e jem p lo , La Tabla 17 p ro p o rc io n a los e jem p los 1 y 2 que p roducen ac ido s grasos. La cepa h o sp e d a d o ra de p ro du cc ion u tilizad a para p ro d u c ir ac ido s g rasos en el e jem p lo 1 es una ce lu la h o sp e d a d o ra de p ro du cc ion que esta g e n o m a n ip u la d a para te n e r las a c tiv id a d e s en z im a tica s s in te ticas deseadas . C a da "x" m arca los genes c o rre la c io n a d o s con las ac tiv ida des , po r e jem p lo , a c tiv id a d e s de a ce til-C o A ca rb ox ilasa , t io e s te ra sa y a c il-C o A s in tasa . Las ce lu las h o sp e d a d o ra s de p ro du cc ion pueden s e le cc io n a rse de bac te rias , le vad u ras y hongos . S egun lo d ispu es to en Tabla 17, u tilizan do los ge ne s e xo g e n o s ind icados, pu ed en c rea rse h o sp e d a d o re s de p ro du cc ion ad ic iona les .

Table 17 p ro po rc io na p ace du p ro du cc ion e jem p la re s ad ic io na le s. S e of sc ribs in two examples of basic routes for the treatment of fatty acids, alcohol and other substances, and those of wax. P o jem p lo, Table 17 p ro po rc io na the e jem p the 1 and 2 that produce fatty acids. The strain ho sp edado ra de p ro du cc ion u tilizad a para p ro duc go ac s sos satos in the e jem p lo 1 is a ce la ho sp edado ra de p ro du cc ion which is genoman ip or the day to have the desired activities in desired techniques. C a da "x" m arca the genes associated with the ac tiv ida des, po re jem p lo, ac tiv id ades de a ce til-C or A ca rb ox ilasa, un tio ra sa yac il-C or A s in rate. They can be used as bac rums, vad u ras and fungi. According to what is stated in Table 17, the use of the indicated genes is possible, in which they can be housed in the ad ditional programs.

EJEMPLO 16. USO DE ACIL-COA SINTASAS ADICIONALES PARA SOBREPRODUCIR ACIL-COA

EXAMPLE 16. USE OF ACIL-COA ADDITIONAL SYNTHESES TO OVERCROW ACIL-COA

H o m o lo g o s a fadD de E. coli pueden e xp re sa rse en E. coli s in te tiza n d o ge ne s o p tim iza d o s po r co d o n e s basandose en una secu en c ia de sea da de M. tuberculosisHR7Rv (N P _ 217021 , F adD D 35), B. subtilis (N P _ 388908 , Y hfL), Saccharomyces cerevisiae (N P _ 012257 , F aa3 p ) o P. aeruginosa PAO 1 (N P _ 251989 ). P ueden d ise n a rse genes s in te tico s para in c lu ir sa lie n te s N co I y HindII com p a tib les . D espues , las a c il-C o A s in ta sas pueden c lo n a rse en un v e c to r pT rcH is2 d ige rido con NcoI/HindIII ( Inv itrog en C orp., C a rlsba d , C a lifo rn ia ) com o se de sc rib e a n te rio rm e n te y exp re sa rse en la cepa M G 1655 AfadE de E. coli. La exp res ion en E. coli puede c o n d u c ir a un a u m en to de la p ro du cc ion de ac il-C oA .

In fact , E. coli fadD can be found in E. coli s in te chalk ndo ge ne sop timiza two po rons based on a section of M. tuberculosisHR7Rv (NP 217021, F adD D 35), B. subtilis (NP_ 388908, Y hfL), Saccharomyces cerevisiae (NP_012257, F aa3 p) or P. aeruginosa PAO 1 (NP_ 251989). It is possible to develop genetic genes in order to go to the United States and India . Then, the ac-C or A s in ta sas can be found in a neighborhood with NcoI / HindIII (Inv itrog in C orp., C a rlsba d, C a lifo rn ia) how r r e sc r ean te rio en te and exp re sa rse in strain MG 1655 AfadE of E. coli. The expiration in E. coli can lead to an increase in the price of ac-C oA.

T a m b ie n pueden p ro du c irse d e riva d o s de ac ido s g rasos, ta l com o FA E E , m ed ia n te la c o tra n s fo rm a c io n de la cepa M G 1655 AfadE de E. coli con d ive rsa s a c il-C o A s in ta sas en el v e c to r pT rcH is2 con un p lasm id o com p a tib le p ro ce d e n te de pC L1920 , que co n tie n e el gen tio e s te r de Cuphea hookeriana con o sin una e s te r s in ta sa de A. baylyi. El h o sp e d a d o r de p ro du cc ion resu ltan te p ro du c ira FA E E cuando se cu ltive en un m ed io que con teng a e tano l com o se de sc rib e a n te rio rm en te .

They can also go from riva two to a few years, as well as FA EE, by way of cooperation with the strain MG 1655 AfadE de E. coli With ive rsa sac il-C or A s in ta sas in the neighbor to r pT rcH is2 with a pasm id or com pa tib le p ro ce den te of pC L1920, which has the uncle gene s te r of Cuphea hookeriana with or without an e s te rs in ta sa of A. baylyi. The ho elder age of the resu ltan te p ro du c will go to FA EE when it is considered in a medium that has as many as r r e sc r ean rio in te.

EJEMPLO 17. USO DE ACIL COA SINTASAS ADICIONALES PARA SOBREPRODUCIR ACIL-COA

EXAMPLE 17. USE OF ACIL COA ADDITIONAL SYNTHESES TO OVERCROW ACIL-COA

S e co n o ce n se cu e n c ia s de A D N o s e cu e n c ia s de p ro te in a s de m uch os h o m o lo g o s de F adD de E. coli. S in em bargo , so lo se han d e sc rito las p ro p ie d a d e s b io q u im ica s de a lg u n a s de e llas . V ease , po r e jem p lo , K no ll e t al., J. Biol. C hem .

It is not known that the DNA is biased or that it is the same as those of many homologues of E. coli. In bargo, however, the reasons for the chemistry of some of them have to be explained. See, po re jem p lo, K no ll et al., J. Biol. C hem.

269 (23 ):16348 -56 , 1994; S h o cke y et al., P lan t P hys io l. 132: 1065 -1076, 2003. A s im ism o , se d e sco n o ce su ca p a c id a d para e xp re sa rse en una fo rm a a c tiva a n ive le s s u fic ie n te m e n te a lto s para fine s com e rc ia les . P ara e xp lo ra r la po s ib ilidad de u tiliza r a c il-C o A s in ta sas h e te ro loga s para la p ro du cc ion de este res, se c lona ron va rio s genes de a c il-C o A s in ta sa y se exp resa ron de la s ig u ie n te m anera . A u n q u e este e jem p lo de sc rib e la tra n s fo rm a c io n del h o s p e d a d o r de p ro du cc ion con p lasm id os d is tin to s para los genes de tio e s te ra sa , e s te r s in ta sa y a c il-C o A s in tasa , es to s genes pueden , com o a lte rna tiva , in co rp o ra rse en un so lo p lasm id o para tra n s fo rm a r el h o sp e d a d o r de p roducc ion .

269 (23): 16348-56, 1994; S h o cke and et al., P lan t P hys io l. 132: 1065-1076, 2003. A s im ism o, I know that it is not possible for you to go on an active annual basis only to mention it for fine s com e rc ia les. For the purpose of the use of racist il-C or A s in ta sas he has made a log for the protection of this area, there are several genes of ac il-C o A s in ta sa and they expired from the same way. Even though this is the reason for the work of the host of the host with the most important criteria for the genes of the uncle, it is the reason, that is rs in ta sa yac il-C or A s in rate, these genes can, as a matter of course, be in the same way for the longest period of time. of production .

1. C o ns trucc ion de l p lasm id o pO P -80

1. C o ns trucc ion de l p lasm id o pO P -80

P ara s o b re e x p re s a r los genes, a tra v e s de la d ige s tion de l p lasm id o p C L1920 con las en z im a s de res tricc ion A flII y S fo I (N e w E ng land B ioLabs Inc. Ipsw ich, MA), se con s tru yo un p lasm id o de ba jo num ero de cop ia s basado en el v e c to r com e rc ia l p C L 1920 (L e rn e r e Inouye, N A R 18: 4631 , 1990 ) que llevab a un p ro m o to r tra n s c r ip c io n a l fue rte . A tra v e s de es ta d ig e s tio n , se c rea ron tre s fra g m e n to s de se c u e n c ia s de A D N . El fra g m e n to de 3737 pb se p u rif ico en ge l u tilizan do un kit de p u rifica c ion en gel (Q iagen , Inc. V a len c ia , CA). En pa ra le lo , un fra g m e n to que con ten ia el p ro m o to r trc y la reg ion lacldel p la sm id o co m e rc ia l pT rcH is2 (Inv itrog en , C a rlsbad , C A ) se a m p lifico m ed ia n te PCR u tiliza n d o los ceb ad o res LF302: 5 '-A T A T G A C G T C G G C A T C C G C T T A C A G A C A -3 '(S E Q ID N O :41); y LF303: 5 '-A A T T C T T A A G T C A G G A G A G C G T T C A C C G A C A A -3 '(S E Q ID N O :42). E stos dos ce b a d o re s ta m b ie n in tro du je ro n s itio s de re c o n o c im ie n to pa ra las e n z im a s de re s tr ic c io n Z ra I (g a c g tc ) y A flII (c ttaag ), re sp e c tiva m e n te , al fina l de los p ro d u c to s de la PCR . D espues de la a m p lifica c io n , los p ro du c to s de la P C R se p u rifica ro n u tilizan do un kit de pu rifica c ion po r P C R (Q iagen , Inc. V a len c ia , C A ) y se d ig ir ie ro n con Z ra I y A flII s ig u ie nd o las con d ic ion es re co m e n d a d a s po r el fa b rica n te (N e w E ng land B io La bs Inc., Ipsw ich, M A). D e sp u e s de la d ige s tion , el p ro du c to de la P C R se pu rifico en ge l y se ligo con el fra g m e n to de 3737 pb de la se cu e n c ia de A D N p ro ce d e n te de pC L1920.

In order to express the genes, through the division of lp lasm id op C L1920 with the areas of res triction A flII and S fo I (N ew E ng land B ioLabs Inc. Ipsw ich, MA), is based on a low copy number based on the neighborhood of CL 1920 (L e rn ere Inouye, NAR 18: 4631, 1990) which led to a transition to the end of the year. Through this scenario, there is a sequence of DNA sequences. The 3737 bp phrase was made available in a gel-based kit (Q iagen, Inc. V a len c ia, CA). For him, a phrase that he had the p rmo mo to r trc and the region lacldel p la sm id o co me rc ia l pT rcH is2 (Inv itrog en, C a rlsbad, CA) PCR was amplified using the primers LF302: 5 '-ATATGACGTCGGCATCCGCTT ACAGACA -3' (SEQ ID NO: 41); and LF303: 5 '-AATTCTTAAGTCAGGAGAGCG TTCACCGACAA -3' (SEQ ID NO: 42). These two countries have been recognized in the same way as the research institutions of the Republic of Mexico (gacg tc) and A flII (c ttaag), re sp ec tiva men te, at the end of the p r ducts of the PCR. After the amplification, the p r du cts of the PCR can be purified by using a PCR kit (Q iagen, Inc. V a len c ia, CA ) and it is agreed with Z ra I and A flII s ig u ie nd or the con d ic ion is recommended by the manufacturer (N ew E ng land B io La bs Inc ., Ipsw ich, MA). Under the direction of the division, the PCR project was able to be certified in general and linked to the 3737 bp sentence of the DNA sequence for DNA. pC tea L1920.

D e sp u e s de la tra n s fo rm a c io n con la m ezc la de ligam ien to en T O P 10 ce lu la s q u im ica m e n te co m p e te n te s (Inv itrogen , C a rlsbad , CA), los tra n s fo rm a n te s se se lecc io na ron en p lacas de a g a r Luria que con ten ian esp e c tin o m ic in a 100 |jg /m l. M uchas co lo n ia s fu e ro n v is ib le s de sp u e s de una in cub ac io n du ran te la noche a 37 °C . Los p lasm idos p re sen te s en es ta s co lo n ia s se pu rifica ron , se a n a liza ro n con e n z im a s de res tricc ion , y d e sp u e s se se cu e n c ia ro n . Se c o n se rvo un p lasm id o p ro du c ido de esta m anera , de no m in a d o pO P -80 , y se u tilizo para e x p e rim e n to s ad ic iona les . En la F igu ra 16 se m u e s tra un m ap a de pO P -80.

T e sp u e s of the tra ns fo rm ac io n with the l e mc la ligam ien to TOP 10 c l lu c i m ica c i m icam te m te te te c te (Inv itrogen, C a rlsbad, CA ), the tram s fo rm an te s were read in Luria agar plates, which had a specific concentration of 100 | jg / ml. Many of them have been part of an in-room at night at 37 ° C. The most important plasmids in this area were only purified, they were added to the rules, and it was known that they were known. A pasm id or p ro du c of this way, of not less than p-80, was used and used for additional purposes. In Fig. 16 a m p a p p P -80 is shown.

S e ve rific a ro n las se cu e n c ia s de A D N de las reg io nes re leva n tes de l p lasm id o pO P -80. S e e n co n tro que en las u n io n e s d o n d e los 2 fra g m e n to s e s ta ban ligad os fa lta ban de 3 a 4 bases en cada extrem o . E sto fue p ro ba b le m en te ca u sa d o po r una ac tiv ida d de e xo n u c le a sa que c o n ta m in a una de las e n z im a s de res tricc ion . E ra p ro ba b le que estas pe qu en as d e le c io n e s no a fe c ta ran n ing una fun c ion im p o rtan te del p lasm ido . El p lasm id o resu ltan te se u tilizo en to d o s los e x p e rim e n to s de exp res ion d e sc rito s en este e jem p lo . La secu en c ia com p le ta de l p lasm ido se de sve la com o S E C ID NO: 1 en la FIG.17.

The consequences of A D N of the relevant regions of the pasm id o pO P -80 are observed. S e e n co n tro that in the n o io n e s d o n d e the 2 frag m e n to s e s ta ta bonded 3 to 4 bases in each end. This was the case in which one of the activities of this period was one of the three months of restitution. E ra p ro ba b le that these peq e ns s e le c io n e s no a fe c ta ran n ing an important function of the plasm. The p lsm id o resu ltan was used in all e x p e rim e n to s of exp res ion d e sc rito s in this e jem p lo. The com p leta section of the pasm gone is seen as S E C ID NO: 1 in FIG. 17.

2. C lona c io n de fadD35 de Tuberculosis micobacteriana H R 7R v

2. C canvas with fadD35 of mycobacterial tuberculosis HR 7R v

A tra ve s de D N A 2.0 Inc. (M en lo Park, CA), se s in te tizo un gen de E. coli o p tim iza d o con codones , u tilizan do com o pun to de pa rtida la secu en c ia de p ro te inas del gen fadD35 d e po s itad a en el N C BI con el num ero de reg is tro N P _217021 de G e n B ank . El gen s in te tico c o n te n ia un s itio N coI e xc lu s ivo en el ex tre m o 5' y un s itio E coR I e xc lu s ivo en el e x trem o 3'. El gen s in te tico se p ro p o rc io n o a tra ve s de D N A 2.0 Inc. c lo n a d o en el p lasm ido pJ201 :16084. El gen fad35 se libe ro de este p lasm id o m ed ia n te d ige s tion con NcoI y EcoR I. La se cu e n c ia de este fra g m e n to se m u e s tra en la S E C ID NO: 2 en la FIG. 18. El fra g m e n to de s e c u e n c ia de A D N re su lta n te q u e se d e sve la en la S E C ID N°: 2 se ligo con pO P -80 , que p re v ia m e n te se d ig irio con NcoI y E coR I. La m ezc la de lig a m ie n to se tra n s fo rm o en ce lu la s T O P 10 qu im ica m e n te com p e ten te s (Inv itrog en , C a rlsbad , CA), que de spu es se co loca ron en p lacas de a g a r Lu ria que con ten ian e sp e c tin o m ic in a 100 pg /m l y se in cubo a 37 °C du ran te la noche . Las co lo n ia s que ap a re c ie ro n al d ia s ig u ie n te se e xp lo ra ro n y se iden tifico una cepa que co n te n ia el p lasm id o co rrec to . El p lasm id o se de no m ino pD S9.

Through DNA 2.0 Inc. (M in Park, CA), an E. coli op tim ized gene was coded with codons, used as the starting point of the sequence in c ia de p ro te ines of the gene fadD35 of p s itad a in the NC BI with the registration number NP _217021 of G in B ank. The genetic system has a positive period in the last 5 'and a critical period in the last 3'. The synthetic gene was spread through DNA 2.0 Inc. c i swam in the pj201: 16084. The gene fad35 Liberty ro this Lasm p id ed om e n I d s ige agement with NcoI and EcoR I. cu enc is ia fra gmen to this tra is Mues in SEQ ID NO: 2 in FIG. 18. The DNA sequence sequence refers to the one in the SEC ID N °: 2 was linked with pO P -80, which p re v ia men te d ig will go with NcoI and E coR I. The linkage m ezc is trans ms fo rm o c ce lu The top 10 chemistry in which you have a lot (Inv itrog en, C a rlsbad, CA), which is then carried out in agaric shells, which had both ic in at 100 pg / ml and was covered at 37 ° C during the night. The co ia ns that ap a re c ie ro n a day s ig u ie n e xp lo ra ro n and a strain was identified that con te nent the pasm id or co r tc. The p id id is not min pD S9.

3. C lona c io n de fadD1 de Pseudomonas aeruginosa PAO1

3. C canvas with fadD1 of Pseudomonas aeruginosa PAO1

A tra ve s de D N A 2.0 Inc. (M en lo Park, CA), se s in te tizo un gen de E. coli o p tim iza d o con codones , u tilizan do com o pun to de pa rtida la secu en c ia de p ro te inas del gen fadDI de p o s ita d a en el N C BI con el n.° de reg is tro N P _ 251989 de G en B ank. El gen s in te tico con ten ia un s itio BspH I un ico en el ex trem o 5 'y un s itio EcoR I un ico en el ex trem o 3'. El gen s in te tico fue p ro p o rc io n a d o a tra v e s de A D N 2.0 , Inc. y se c lo no en el p la sm id o p J201 :16083. El gen fadDI se libe ro de este p lasm id o m ed ia n te d ige s tion con BspH I y E coR I. La secu en c ia de este fra g m e n to se m ue s tra en la S E C ID NO: 3 en la FIG . 19. El fra g m e n to de se cu e n c ia de A D N resu ltan te de S E Q ID NO: 3 se ligo con pO P -80, que p re v ia m e n te se d ig irio con NcoI y E coR I. La m ezc la de ligam ien to se tra n s fo rm o en ce lu la s T O P 10 q u im ica m e n te co m p e te n te s (Inv itrog en , C a rlsbad , CA), que d e sp u e s se co loca ron en p lacas de a g a r Lu ria que co n ten ian e sp e c tin o m ic in a 100 pg /m l y se incubo a 37 °C d u ran te la noche. Las co lon ias que a p a re c ie ro n al d ia s ig u ie n te se exp lo ra ron . S e id en tifico una cepa que co n te n ia el p la sm id o co rrecto . El p lasm id o se d e n o m in o pD S8.

Through DNA 2.0 Inc. (M in Park, CA), an E. coli op tim ized gene was coded with codons, used as the starting point of the sequence in c ia de p ro te ines of the fadDI gene of pos ita da in NC BI with the registration number NP _ 251989 of G in B ank. The genetic group had a unique BspH I site in the last 5 'and a unique EcoR I site in the last 3'. The genetic technique was carried out by DNA 2.0, Inc. and was reported in the J201: 16083. The fadDI gene was released from this plasm id om ed ia n te d ige s tion with BspH I and E coR I. The section of this fragrance is shown in SEQ ID NO: 3 in FIG. 19. The statement of the summary DNA of SEQ ID NO: 3 was linked with pO P -80, which, in turn, went with NcoI and E coR I. This is the link between the top 10 chemistry and men (Inv itrog en, C a rlsbad, CA), which is co They rummed in agar plates, which they had at 100 pg / ml and incubated at 37 ° C overnight. The communities that appeared at the same time were only explained to you. A strain was identified in which the correct name was correct. The plasm id is called in or pD S8.

4. C lona c io n de yhfL de Bacillus subtilis

4. C yhfL canvas of Bacillus subtilis

El gen yhfL se a m p lifico po r P C R u tiliza n d o B a c illu s subtilis La secu en c ia de A D N c ro m o s o m ic o I168 com o m o lde , y dos ce b a d o re s d isen ado s en base a la secu en c ia de A D N de p o s ita d a en N C BI con el num ero de acce so de G e n B a n k N C _000964. Las s e cu e n c ia s de los 2 c e b a d o re s fue ron :

The yhfL gene was amplified by PCR or used as a subtilis. The sequencing of the DNA as a whole or as an I168, and two primed re sd ised based on the secu in the case of DNA of posi- tion in NC BI with the accession number of G in B ank NC _000964. The questions of the 2 priming were:

B syh fL B spH IF : 5 '-C A T C A T G A A T C T T G T T T C -3 ' (S E Q ID N O :4) (FIG. 20)

B syh fL B spH IF: 5 '-C A T C A T G A A T C T T G T T T C -3' (S E Q ID N O: 4) (FIG. 20)

B syh fL E coR : 5 '-C G G A A T T C T T A T T G G G G C A A A A T A T C -3 ' (S E Q ID N O :5 ) (FIG. 21)

B syh fL E coR: 5 '-C G G A A T T C T T A T T G G G G C A A A A T A T C -3' (S E Q ID N O: 5) (FIG. 21)

E stos dos ce b a d o re s in tro du je ro n un sitio de re co n o c im ie n to B spH I en el e x trem o 5 'y un sitio de reco n o c im ie n to E coR I en el ex tre m o 3'. El p ro du c to de la P C R se c lono d ire c ta m e n te en el v e c to r pC R -B lu n t II-T O P O u tiliza n d o el kit de c lo n a c io n de P C R Z e ro B lu n t T O P O (Inv itrog en , C a rlsba d , C A). A l p lasm id o que llevab a el gen yhfL se le d e n o m in o pD S 1. P ara s u b c lo n a r el gen yhfL, el p lasm ido pDS1 se d ig irio con BspH I y E coR I. El fra g m e n to de se cu e n c ia de A D N re su lta n te S E Q ID NO: 6 (FIG. 22) se pu rifico en gel y se c lono en pO P -80 , que p re v ia m e n te se d ig irio con N coI y E coR I. A l p lasm id o que llevab a el gen yhfL de B. subtilis c lo n a d o en pO P -80 se le d e n o m in o pD S4.

These two are in tro du je ro n a site of recognition sp B I in the last 5 'and a site of recognition in the past I in the ex tre mo 3 '. The p r du c t of the PCR was extended directly in the neighboring pC R-B lu nt II-T OPO or the kit used for the PCRZ e ro B lu nt TOPO (Inv itrog en, C a rlsba d, CA). The term that leads to the yhfL gene is referred to as pD S 1. In order to raise the yhfL gene , the plasmid pDS1 was given with BspH I and E coR I. The phrase According to DNA, its SEQ ID NO: 6 (FIG. 22) was gel gelled and was pO P -80, which was given with N coI and E coR I. The term that leads to the yhfL gene of B. subtilis c swim in pO P -80 is called in or pD S4.

5. C lona c io n de faa3p de Saccharomyces cerevisiae (N P 012257 )

5. C tarpaulin of faa3p from Saccharomyces cerevisiae (NP 012257)

El gen faa3p gen se a m p lifico po r P C R u tilizan do com o m o lde la secu en c ia de A D N c ro m o so m ico co m e rc ia l de Saccharomyces cerevisiae A T C C 204508 D (A m e rica n T yp e C u ltu re C o lle c tion , M an assa s , V A ) y dos c e b a d o re s que se d ise n a ro n b a san do se en la secu en c ia de A D N de p o s ita d a en el N C BI con el n.° de reg is tro N C _001141 de G en B ank. Las s e cu e n c ia s de los dos c e b a d o re s fue ron :

The faa3p gene was amplified by PCR or used as a DNA sequence by Saccharomyces cerevisiae ATCC 204508 D (A me rica n T yp e C u l ct re c o c tion, M an assa s, VA) and two re priming that is given in ro nba san do se in the sequencing of DNA of position in NC BI with the n Reg. N ° NC _001141 of G in B ank. The questions of the two primers were:

S c faa 3pP c iF : 5 '-C G A C A T G T C C G A A C A A C A C -3 ' (S E Q ID N O :7 ) (FIG. 23)

S c faa 3pP c iF: 5 '-C G A C A T G T C C G A A C A A C A C -3' (S E Q ID N O: 7) (FIG. 23)

S c faa 3pP c iI: 5 '-G C A A G C T T C T A A G A A T T T T C T T T G -3 ' (S E Q ID N O :8) (FIG. 24)

S c faa 3pP c iI: 5 '-G C A A G C T T C T A A G A A T T T T C T T T G -3' (S E Q ID N O: 8) (FIG. 24)

E stos dos ce b a d o re s in tro du je ro n en un s itio de re co n o c im ie n to Pci I en el ex tre m o 5' y en un s itio de re co n o c im ie n to H ind III en el ex tre m o 3'.

These two re s in tro du je ro n in a reCoC noc im ie to Pci I in the last 5'm and in a reCoC noc im ie in to H ind III in the last tre 3 '.

El p ro du c to de la P C R se c lo no d ire c ta m e n te en el v e c to r p C R -B lu n t II-T O P O u tilizan do el k it de c lo n a c io n de PC R Z e ro B lu n t T O P O (Inv itrog en , C a rlsbad , CA). A l p lasm id o que llevaba el gen faa3p se le de no m in o pD S2. Para s u b c lo n a r el gen faa3p, el p lasm id o pD S 2 se d ig ir io con PciI y H ind III. El fra g m e n to de se cu e n c ia de A D N (S E Q ID NO: 9) (FIG. 25) se p u rif ico en ge l y se c lo n o en pO P -80 , qu e p re v ia m e n te se d ig ir io con N coI y H ind III. A l p la sm id o que llevab a el gen faa3p de S. ce re v is ia e c lo na do en pO P -80 se le d e n o m in o pD S5.

The p r du c t of the PCR is c ndirect c ta men t in the neighbor CR CR-B lu nt II-T OPO or use the c k it of c r born of PC RZ e ro B lu nt TOPO (Inv itrog en, C a rlsbad, CA). Lp lasm id o that carried the faa3p gene is given no m in or pD S2. To subclass the faa3p gene , the plasm id or pD S 2 was given with PciI and H ind III. The DNA sequencing phrase (SEQ ID NO: 9) (FIG. 25) was published in general and in pO P -80, which is reported ig went with N coI and H ind III. Lp la sm id o that led to the faa3p gene of S. ce re v is ia ec lo na do in pO P -80 is called in or pD S5.

6. C lona c io n de ZP 01644857 de Stenotrophomonas maltofila R 551 -3

6. C canvas with ZP 01644857 from Stenotrophomonas maltofila R 551 -3

La secu en c ia gen ica es tru c tu ra l de la p ro te ina Z P _ 01644857 esta d ispo n ib le en el N C BI com o pa rte del locus N Z _ A A V Z 01000044. El gen se a m p lifico po r P C R u tilizan do com o m o lde la se cu e n c ia de A D N c ro m o so m ico de Stenotrophomonas maltofila R 551 -3 y dos ce b a d o re s d ise n a d o s b a san do se en la se cu e n c ia de A D N de po s itad a . Las s e cu e n c ia s de los dos ce b a d o re s fue ron :

The gene sequence is tru c tu ra l de la p ro te ina ZP _ 01644857 is available in the NC BI as part of the locus NZ _ AAVZ 01000044. The gene was amplified by r PCR uses the DNA sequence as the only form of Stenotrophomonas maltofila R 551 -3 and two primed re sd ise n adosba san do se in the DNA of pse itad a. The questions of the two fathers were:

S m p rk59 B sp F : 5 '-A G T C A T G A G T C T G G A T C G -3 ' (S E Q ID N O :10 ) (FIG. 26)

S m p rk59 B sp F: 5 '-A G T C A T G A G T C T G G A T C G -3' (S E Q ID N O: 10) (FIG. 26)

S m p rk59 H in d R : 5 '-G G A A G C T T A C G G G G C G G G C G -3 ' (S E Q ID N O :11 ) (FIG. 27)

S m p rk59 H in d R: 5 '-G G A A G C T T A C G G G G C G G G C G -3' (S E Q ID N O: 11) (FIG. 27)

E stos dos ceb ad o res in tro du je ro n un s itio de re co n o c im ie n to B spH I en el ex trem o 5' y en un s itio de reco no c im ie n to H ind III en el ex tre m o 3'.

These two ceb ad ores in tro du je ro n a forum of re co noc im ie n to B spH I in the last 5 'and in a collection site no c im ie n to H ind III in the last tre 3 '.

El p ro du c to de la P C R se c lo no d ire c ta m e n te en el v e c to r p C R -B lu n t II-T O P O u tilizan do el k it de c lo n a c io n de PC R Z e ro B lu n t T O P O (Inv itrog en , C a rlsbad , CA). A l p lasm id o que llevab a el gen que cod ifica la p ro te ina Z P _ 01644857 se le de no m ino pD S3. P ara fa c ilita r la sub c lo nac ion ad ic io na l del gen, se e lim ino un s itio in te rno de B spH I m ed ian te m u ta g e n e s is d irig ida u tilizan do el c e b a d o r P rkB sp: 5 '-G C G A A C G G C C T G G T C T T T A T G A A G T T C G G T G G -3 '(S E Q ID NO: 12) (FIG. 28) y el k it de m u ta g e n e s is d irig ida a m u ltip le s s itio s de Q u ikC h a n g e (S tra tag ene , La Jo lla , CA). D e sp u e s de co rro b o ra r la m u tac ion a p ro p ia d a po r se cu e n c ia c io n de A D N , el p lasm id o resu ltan te se d ig ir io con BspH I y H indIII, y se de no m ino pD S 6. El fra g m e n to