ES2665879T3 - Método para producir aceite biológico usando un fermentador no estéril - Google Patents
Método para producir aceite biológico usando un fermentador no estéril Download PDFInfo
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- ES2665879T3 ES2665879T3 ES08830498.5T ES08830498T ES2665879T3 ES 2665879 T3 ES2665879 T3 ES 2665879T3 ES 08830498 T ES08830498 T ES 08830498T ES 2665879 T3 ES2665879 T3 ES 2665879T3
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- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6436—Fatty acid esters
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- C12P7/6458—Glycerides by transesterification, e.g. interesterification, ester interchange, alcoholysis or acidolysis
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- C12P7/00—Preparation of oxygen-containing organic compounds
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Abstract
Un método para producir un aceite biológico que comprende cultivar un microorganismo del reino Stramenopile, en el que dicho microorganismo es un traustoquítrido, seleccionado de los géneros Schizochytrium, Thraustochytrium y Ulkenia, por fermentación heterotrófica, en el que más de un 50 % a un 99 % de los ácidos grasos insaturados en dicho aceite biológico son ácidos grasos poliinsaturados, y en el que dicha fermentación se realiza en un fermentador no estéril.
Description
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producción de lípidos para producir un aceite biológico; y (b) craquear el aceite biológico para producir biocombustible de aviones.
Breve descripción de los dibujos
La figura 1 muestra diversas realizaciones de un método de producción de aceites biológicos y biodiésel de acuerdo con la presente invención. La figura 2 muestra un ejemplo de un diseño de sistema de fermentación de acuerdo con la presente invención. La figura 3 muestra gráficos del peso celular seco, el porcentaje en peso de lípidos, el porcentaje en peso de DHA y la cantidad de lípidos producida por litro de caldo de fermentación durante el tiempo para el crecimiento de un microorganismo (ATCC 20888) en condiciones estériles y no estériles descritas en el ejemplo 4. La figura 4 muestra gráficos de la tasa de consumo de azúcar, la tasa de producción de aceite (como gramos por litro de caldo de fermentación por día), la tasa de productividad de biomasa (en gramos por litro por día) y la cantidad de biomasa sin lípidos durante el tiempo para el crecimiento de un microorganismo (ATCC 20888) en condiciones estériles y no estériles descritas en el ejemplo 4. La figura 5 muestra un diagrama de un proceso de fermentación de dos fases que comprende una fase de siembra continua y una fase de acumulación de lípidos discontinuo.
Descripción detallada de la invención
La presente invención proporciona un método para producir un aceite biológico, que comprende cultivar un microorganismo del reino Stramenophile donde dicho microorganismo es un traustoquítrido, seleccionado de los géneros Schizochytrium, Thraustochytrium, y Ulkenia, por fermentación heterotrófica, donde más de un 50 % a un 99 % de los ácidos grasos insaturados en dicho aceite biológico son ácidos grasos poliinsaturados, y donde dicha fermentación se realiza en un fermentador no estéril. Algunas realizaciones del método de la presente invención proporcionan organismos heterotróficos oleaginosos y procesos adecuados para convertir el carbono, mediante fermentación, de sacáridos directamente basados en celulosa o basados en lignocelulosa en aceite vegetal para la fabricación de biodiésel. Los procesos de la presente invención tendrían mayor capacidad de cambio de escala, más sostenibles y generarían un biodiésel más competitivo en costes que los procesos actualmente usados o investigados (tal como biodiésel de aceite de semillas o biodiésel de algas fotosintéticas).
En este documento se divulga además el crecimiento de alta densidad de dos microorganismos oleaginosos usando materias básicas celulósicas sacarificadas. Por ejemplo, el protista Schizochytrium sp y la levadura oleaginosa Yarrowia lipolytica son adecuados para dichos procesos porque ambos tienen sistemas de transformación bien desarrollados para modificar los microorganismos y pueden producir altos niveles de lípidos por fermentación. También se divulgan traustoquítridos oleaginosos y hongos que pueden crecer en una diversidad de sustratos celulósicos y lignocelulósicos, y organismos que pueden ser susceptibles de forma natural a sacarificación y fermentación combinadas, así como degradación de lignina o resistencia.
La presente invención también puede usar cepas mejoradas de los microorganismos de la reivindicación 1 y procesos para utilizar sustratos basados en celulosa para la producción de aceite mediante medios moleculares, biológicos, genéticos clásicos y fisiológicos. Algunas realizaciones de la presente invención pueden proporcionar un aumento de escala económico de un proceso de fermentación para convertir acilglicéridos celulares en biodiésel. En este documento se divulgan diseños de reactor biológico y químico y construcciones, así como estrategias de producción comercial para la implementación de los métodos de la invención.
En la presente divulgación, los organismos incluyen aquellos seleccionados del grupo que consiste en algas doradas (tales microorganismos del reino Stramenopiles), algas verdes, diatomeas, dinoflagelados (tales como microrganismos del orden Dinophyceae, incluyendo miembros el género Crypthecodinium tal como, por ejemplo, Crypthecodinium cohnii), levaduras tal como un miembro de los géneros Yarrowia (tales como Yarrowia lipolytica), Cryptococcus (tal como Cryptococcus albidus), Trichosporon, Candida, Lipomyces, Rhodosporidium, y Rhodotorula), y hongos de los géneros Mucor y Mortierella, incluyendo, aunque sin limitación Mortierella alpina y Mortierella sect. schmuckeri. Los miembros del grupo microbiano Stramenopiles incluyen microalgas y microorganismos de tipo alga, incluyendo los siguientes grupos de microorganismos: Hamatores, proteromónadas, opalinos, Devalpayella, Diplophrys, labrintúlidos, traustoquítridos, biosécidos, oomicetos, hipoquitridiomicetos, Commation, Reticulospahaera, pelagomonas, Pelagococcus, Ollicola, Aureococcus, parmales, diatomeas, xantofitos, feofitos (algas pardas), eustigmatofitos rafidofitos, sinúridos, axodinos (incluyendo Rhizochromulinaales, Pedinellales, Dictyochales), crisomeridales, sarcinocrisidales, hidrurales, hiberdiales y cromulinales. Los traustoquítridos incluyen los géneros Schizochytrium (las especies incluyen aggregatum, limnaceum, mangrovei, minutum, octosporum), Thraustochytrium (las especies incluyen arudimentale, aureum, benthicola, globosum, kinnei, motivum, multirudimentale, pachydermum, proliferum, roseum, striatum), Ulkenia (las especies incluyen amoeboidea, kerguelensis, minuta, profunda, radiate, sailens, sarkariana, schizochytrops, visurgensis, yorkensis), Aplanochytrium (las especies incluyen haliotidis, kerguelensis, profunda, stocchinoi), Japonochytrium (las especies incluyen marinum), Althornia (las especies incluyen crouchii)y Elina (las especies incluyen marisalba, sinorifica). Los labrintúlidos incluyen los géneros Labyrinthula (las especies incluyen algeriensis, coenocystis, chattonii, macrocystis, macrocystis atlantica, macrocystis macrocystis, marina, minuta, roscoffensis, valkanovii, vitellina, vitellina pacifica,
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reflujo de los componentes de reacción. Por ejemplo, realizar la etapa de reacción de la composición de aceite a presiones mayores de la presión atmosférica puede aumentar el punto de ebullición de los disolventes presentes en la mezcla de reacción. En dichas condiciones, la reacción puede producirse a una temperatura en que los disolventes hervirían a presión atmosférica, pero no provocaría el reflujo de los componentes de reacción. En algunas realizaciones, la reacción se realiza a una presión de aproximadamente 5 a aproximadamente 20 libras por pulgada cuadrada, psi (aproximadamente 6895 a aproximadamente 137 895 Pa); de aproximadamente 7 a aproximadamente 15 psi (aproximadamente 48 263 a aproximadamente 103 421 Pa); o de aproximadamente 9 a aproximadamente 12 psi (aproximadamente 62 052 a aproximadamente 82 737 Pa). En determinadas realizaciones, la reacción se realiza a una presión de 7, 8, 9, 10, 11 o 12 psi (42 263, 55 158, 62 052, 68 947, 75 842 o 82 737 Pa). Las reacciones realizadas a presión pueden realizarse a las temperaturas de reacción enumeradas anteriormente. En algunas realizaciones, las reacciones realizadas a presión pueden realizarse a al menos aproximadamente 70 ºC, 75 ºC, 80 ºC, 85 ºC o 90 ºC. En algunas realizaciones, las reacciones realizadas a presión pueden realizarse a 70 ºC, 75 ºC, 80 ºC, 85 ºC o 90 ºC.
La mezcla de reacción que comprende ésteres de ácido graso puede procesarse adicionalmente para obtener los ésteres de ácido graso de la mezcla. Por ejemplo, la mezcla puede enfriarse, diluirse con agua y la solución acuosa extraerse con un disolvente tal como hexano para producir una composición que comprende ésteres de ácido graso. Las técnicas para lavar y/o extraer mezclas de reacción en bruto son conocidas en la técnica.
En algunas realizaciones de la presente invención, se usan microorganismos que producen bajos niveles de PUFA para producir los aceites biológicos, especialmente para su uso en producción biodiésel. Este método podría reducir los costes de producción de biodiésel.
Los ésteres de PUFA más valiosos pueden recuperarse por destilación para producir ésteres de PUFA de alta potencia que después pueden venderse para reducir el coste global de la producción del producto de biodiésel.
Los ejemplos y sistemas de producción de lípidos modificados se divulgan en las publicaciones de solicitud de patente n.º WO 06/031699, US 2006/0053515, US 2006/0107348 y WO 06/039449.
En una realización de la presente invención, los ésteres de ácido graso se separan de la mezcla de reacción destilando la composición para recuperar una fracción que comprende el éster del ácido graso. De esta manera, una fracción diana de la mezcla de reacción que incluye los ésteres de ácido graso de interés puede separarse de la mezcla de reacción y recuperarse.
En determinadas realizaciones, la destilación se realiza al vacío. Sin limitarse a teoría alguna, la destilación al vacío permite que la destilación se consiga a una temperatura inferior que en ausencia de un vacío y, por tanto, puede evitar la degradación de los ésteres. Las temperaturas típicas de destilación varían de aproximadamente 120 ºC a aproximadamente 170 ºC. En algunas realizaciones, la etapa de destilación se realiza a una temperatura de menos de aproximadamente 180 ºC, menos de aproximadamente 175 ºC, menos de aproximadamente 170 ºC, menos de aproximadamente 165 ºC, menos de aproximadamente 160 ºC, menos de aproximadamente 155 ºC, menos de aproximadamente 150 ºC, menos de aproximadamente 145 ºC, menos de aproximadamente 140 ºC, menos de aproximadamente 135 ºC o menos de aproximadamente 130 ºC. Las presiones típicas para destilación al vacío varían de aproximadamente 0,1 mm de Hg a aproximadamente 10 mm de Hg (aproximadamente 13,3 a aproximadamente 133 Pa). En algunas realizaciones, la presión para la destilación al vacío es de al menos aproximadamente 0,1, 0,5, 1, 1,5, 2, 2,5, 3, 3,5 o 4 mm de Hg (13,3, 66,66, 133,33, 199,98, 266,65, 333,30, 399,96, 466,63 o 533,29 Pa). En algunas realizaciones de la presente invención, la presión para la destilación al vacío es de aproximadamente 0,1, 0,5, 1, 1,5, 2, 2,5, 3, 3,5 o 4 mm de Hg (13,3, 66,66, 133,33, 199,98, 266,65, 333,30, 399,96, 466,63 o 533,29 Pa).
En algunas realizaciones de la presente invención los ésteres de ácidos grasos producidos a través de transesterificación de los aceites biológicos se aíslan adicionalmente a través de aducción de urea. La urea puede disolverse en un medio que comprende los ésteres de ácido graso para formar un medio que comprende ésteres de ácido graso y urea disuelta. Este medio entonces se enfría o concentra para formar un precipitado que comprende urea y al menos una parte de los ésteres de ácido graso saturados y una fracción líquida que comprende al menos la mayoría de los ésteres de ácido graso poliinsaturados. El precipitado y la fracción líquida entonces puede separarse para aislar los ésteres de ácido graso saturados o poliinsaturados. En algunas realizaciones de la presente invención, el medio que comprende los ésteres de ácido graso y la urea disuelta se enfría hasta una temperatura de aproximadamente 20 ºC a aproximadamente -50 ºC, de aproximadamente 10 ºC a aproximadamente -40 ºC o de aproximadamente 0 ºC a aproximadamente -30 ºC. La patente de Estados Unidos n.º 6.395.778 divulga métodos de transesterificación seguida por aducción de urea.
Además de los métodos de transesterificación descritos anteriormente, también pueden incorporarse otras técnicas de reducción de la viscosidad de los aceites bilógicos producidos usando la presente invención en los métodos de la invención para producir biocombustibles basados en lípidos. Estas técnicas incluyen, aunque sin limitación, el uso de lipasas, catálisis de metanol supercrítico y el uso de sistemas de células completas que implican sobreexpresión citoplasmática de lipasas en una célula hospedadora seguida por permeabilización del hospedador para permitir la
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KR20100067111A (ko) | 2010-06-18 |
NZ584460A (en) | 2012-10-26 |
IL204479A (en) | 2017-01-31 |
EA019387B1 (ru) | 2014-03-31 |
US20090064567A1 (en) | 2009-03-12 |
KR20160052805A (ko) | 2016-05-12 |
AR068429A1 (es) | 2009-11-18 |
JP2010538642A (ja) | 2010-12-16 |
US20190177754A1 (en) | 2019-06-13 |
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AU2008300002B2 (en) | 2014-04-10 |
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US11104923B2 (en) | 2021-08-31 |
EP2198038A4 (en) | 2012-04-18 |
CA2699406C (en) | 2019-09-03 |
AU2008300002A1 (en) | 2009-03-19 |
CL2008002699A1 (es) | 2009-08-28 |
EP2198038B1 (en) | 2018-03-14 |
JP2015002743A (ja) | 2015-01-08 |
EA201000470A1 (ru) | 2010-10-29 |
CN113846129A (zh) | 2021-12-28 |
CN101874117A (zh) | 2010-10-27 |
US10308965B2 (en) | 2019-06-04 |
BRPI0815860B1 (pt) | 2021-04-20 |
ZA201002276B (en) | 2012-09-26 |
US9453172B2 (en) | 2016-09-27 |
WO2009035551A1 (en) | 2009-03-19 |
EP2198038A1 (en) | 2010-06-23 |
BRPI0815860A2 (pt) | 2020-06-09 |
NZ598199A (en) | 2013-12-20 |
CA2699406A1 (en) | 2009-03-19 |
MX2010002825A (es) | 2010-08-31 |
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