Микроорганизм, имеющий повышенную продуктивность в отношении молочной кислоты, и способ получения молочной кислоты с использованием данного микроорганизма

FIELD: biotechnology.SUBSTANCE: modified microorganism is characterized by the fact that it reduces the activity of pyruvate decarboxylase, increases the activity of aldehyde dehydrogenase and acetyl-CoA synthetase, and introduces lactic acid dehydrogenase. As a result of these modifications, this microorganism is able to produce lactic acid in high yield. The invention also relates to a process for the production of lactic acid. The method comprises culturing said microorganism and isolating from the culture medium lactic acid produced by culturing this microorganism.EFFECT: production of lactic acid in high yield.6 cl, 1 dwg, 14 tbl, 10 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 636 467 C2 (51) МПК C12N 1/18 (2006.01) C12N 15/52 (2006.01) C12P 7/56 (2006.01) C12R 1/865 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (21)(22) Заявка: 2015130528, 08.05.2015 (24) Дата начала отсчета срока действия патента: 08.05.2015 Дата регистрации: Приоритет(ы): (30) Конвенционный приоритет: 09.05.2014 KR 10-2014-0055865 (45) Опубликовано: 23.11.2017 Бюл. № 33 (73) Патентообладатель(и): СиДжей ЧеилДжеданг Корпорейшн (KR) (85) Дата начала рассмотрения заявки PCT на национальной фазе: 16.11.2015 (56) Список документов, цитированных в отчете о поиске: US 8071357 B2, 06.12.2011. RU 2268304 C1, 20.01.2006. EP 2281881 A1, 09.02.2011. US 20100062505 A1, 11.03.2010. (86) Заявка PCT: KR 2015/004600 (08.05.2015) (87) Публикация заявки PCT: 2 6 3 6 4 6 7 (43) Дата публикации заявки: 17.05.2017 Бюл. № 14 R U 23.11.2017 (72) Автор(ы): ЯНГ Эн Бин (KR), ЛИ Тэ Хи (KR), КИМ Сон Хе (KR), СОНГ Гю Хён (KR), ХА Чол Воонг (KR), НА Кюнг Су (KR), ЯНГ Юнг Леол (KR), КАНГ Мин Сун (KR), ЛИ Хё Хён (KR) 2 6 3 6 4 6 7 R U Адрес для переписки: 191036, г. Санкт-Петербург а/я 24 'НЕВИНПАТ' (54) Микроорганизм, имеющий повышенную продуктивность в отношении молочной кислоты, и способ получения молочной кислоты с использованием данного микроорганизма (57) Реферат: ...

СПОСОБ ПОЛУЧЕНИЯ АЦЕТОИНА

Микроорганизм, продуцирующий молочную кислоту, и способ продуцирования молочной кислоты с его использованием

YEAST ORGANISM PRODUCING ISOBUTANOL AT A HIGH YIELD

There is disclosed a method of producing isobutanol. In an embodiment, the method includes providing a microorganism transformed with an isobutanol producing pathway containing at least one exogenous gene. The microorganism is selected to produce isobutanol from a carbon source at a yield of at least 10 percent theoretical. The method includes cultivating the microorganism in a culture medium containing a feedstock providing the carbon source, until isobutanol is produced. The method includes recovering the isobutanol. In one embodiment, the microorganism is a yeast with a Crabtree-negative phenotype. In another embodiment, the microorganism is a yeast microorganism with a Crabtree-positive phenotype. There is disclosed a microorganism for producing isobutanol. In an embodiment, the microorganism includes an isobutanol producing pathway containing at least one exogenous gene, and is selected to produce a recoverable quantity of isobutanol from a carbon source at a yield of at least 10 percent ...

METHOD FOR THE PREPARATION OF 1,3-PROPANEDIOL FROM SUCROSE

The present invention concerns a microorganism genetically modified for the bioproduction of 1,3-propanediol from sucrose,wherein the microorganism comprises: -a two-step metabolic pathway for the production of 1,3-propanediol, comprising a first step of decarboxylation of4-hydroxy-2-ketobutyrate with an enzyme having a 2-keto acid decarboxylase activity, and a second step of reduction of the obtained 3-hydroxypropionaldehyde with an enzyme having hydroxy aldehyde reductase activity,and -genes enabling the microorganism to utilize sucrose as sole carbon source. The invention also concerns a new method for the biological preparation of 1,3- propanediol by fermentation,comprising cultivating said microorganism genetically modified, wherein the culture is performed in an appropriate medium comprising a source of sucrose, and recovering the 1,3-propanediol being produced. In a preferred aspect of the invention, the source of sucrose is obtained from plant biomass.

ACETATE SUPPLEMENTION OF MEDIUM FOR BUTANOLOGENS

The invention relates to the fields of industrial microbiology and alcohol production. More specifically, the invention relates to improved production of butanol isomers by recombinant microorganisms containing an engineered butanol pathway and disrupted activity of the genes in pathways for the production of by-products during the fermentation when the microorganisms are grown in a fermentation medium containing acetate. In embodiments, recombinant microorganisms have an increased growth rate in a fermentation medium containing acetate as a C2 supplement.

RECOMBINANT STRAIN OFCANDIDASPP., AND THE PREPARATION PROCESS AND USES THEREOF

Yeast cell having acid tolerance, method of preparing yeast cell and use thereof

Provided are a genetically engineered yeast cell having increased activity of SUL1, STR3, HXT7, ERR1, GRX8, MXR1, GRE1, MRK1, AAD10 or a combination thereof, compared to a parent cell, and also having acid tolerance, a method of preparing the same, and a method of producing lactate using the same.

CONSTRUCTION OF A LACTOBACILLUS CASEI ETHANOLOGEN

An engineered bacterium for producing ethanol from one or more carbohydrates is disclosed. The bacterium can be made by (a) inactivating within a bacterium one or more endogenous genes encoding a lactate dehydrogenase; or (b) introducing into a bacterium one or more exogenous genes encoding a pyruvate decarboxylase and one or more exogenous genes encoding an alcohol dehydrogenase II; or (c) performing both steps (a) and (b). The resulting engineered bacterium produces significantly more ethanol than the wild-type bacterium, and can be used in producing ethanol from a substrate such as biomass that includes carbohydrates. 1. A method of making ethanol comprising:{'i': Lactobacillus casei', 'L. casei, 'culturing on a substrate comprising a carbohydrate an engineered bacterium comprising a bacterium that includes one or more exogenous genes encoding a pyruvate decarboxylase and one or more exogenous genes encoding an alcohol dehydrogenase II, each of which is operably linked to a promoter that is highly expressed in the stationary phase, whereby the engineered bacterium produces a composition comprising ethanol.'}2. The method of claim 1 , wherein the engineered bacterium further includes one or more gene deletion mutations of one or more endogenous genes encoding a lactate dehydrogenase.3. The engineered bacterium of claim 1 , wherein the engineered bacterium further includes a gene deletion mutation of an endogenous gene encoding D-hydroxyisocaproate dehydrogenase.5. The method of claim 2 , wherein the gene deletion mutations comprise A L-lactate dehydrogenase 1 (ΔL-ldh1).5. The method of claim 2 , wherein the gene deletion mutations comprise A L-lactate dehydrogenase 2 (ΔL-ldh2).6. The method of claim 1 , wherein the engineered bacterium includes the gene deletion mutations Δ D-lactate dehydrogenase (ΔD-ldh) or Δ D-hydroxyisocaproate dehydrogenase (ΔD-hic).7Zymomonas mobilisZymomonas mobilis. The method of claim 1 , wherein the exogenous gene encoding a pyruvate ...

Increased production of isobutanol in yeast with reduced mitochondrial amino acid biosynthesis

Yeast cells with reduced activity of certain enzymes involved in branched chain amino acid biosynthesis in yeast mitochondria are described. Target enzymes include threonine deaminase, isopropylmalate synthase, and optionally branched chain amino acid transaminase.

Electron consuming ethanol production pathway to displace glycerol formation in S. cerevisiae

The present invention provides for a mechanism to completely replace the electron accepting function of glycerol formation with an alternative pathway to ethanol formation, thereby reducing glycerol production and increasing ethanol production. In some embodiments, the invention provides for a recombinant microorganism comprising a down-regulation in one or more native enzymes in the glycerol-production pathway. In some embodiments, the invention provides for a recombinant microorganism comprising an up-regulation in one or more enzymes in the ethanol-production pathway.

Bacterial cocultures expressing a bacteriocin system

The present disclosure concerns a co-culture of bacterial cells for making a fermented product from a biomass. The co-culture comprising a first recombinant lactic acid bacteria (LAB) cell expressing at least one bacteriocin and a second recombinant lactic acid bacteria (LAB) cell capable of converting, at least in part, the biomass into the fermented product. The second recombinant LAB cell is immune to the bacteriocin produced by the first recombinant LAB cell. The co-culture can be used, optionally in combination with a yeast host cell, to make a fermented product. The present disclosure also provides processes for making the fermented product by using the co-culture as wells kits and media comprising the co-culture.

Lyase und für die Lyase kodierende DNA, die DNA enthaltende Vektoren, sowie Verfahren zur asymmetrischen Synthese von (S)-Phenylacetylcarbinol

Die Erfindung betrifft eine Lyase und eine für die Lyase kodierende DNA, die DNA enthaltende Vektoren sowie ein Verfahren zur asymmetrischen Synthese von (S)-Phenylacetylcarbinol. Erfindungsgemäß wird eine Lyase zur Verfügung gestellt, bei der Trypotophan in dem gegenüber dem Wildtyp aus Aceobacter pasteurianus veränderten Protein ApPDC-E469G in Position 543 durch eine Aminosäure ersetzt ist, bzw. weniger raumerfüllend als Tryptophan ist. Weiterhin werden erfindungsgemäß Desoxyribonukleinsäuren zur Verfügung gestellt, die für die Lyase kodieren. Mit der erfindungsgemäßen Lyase kann (S)-Phenylacetyl-carbinol aus den Edukten Benzaldehyd und Pyruvat bzw. Acetaldehyd mit einem Enantiomerenüberschuss von mindestens 94% hergestellt werden.

Lyase sowie Verfahren zur asymmetrischen Synthese von (S)-Phenylacetylcarbinol

Die Erfindung betrifft eine Lyase sowie Verfahren zur asymmetrischen Synthese von (S)-Phenylacetylcarbinol Erfindungsgemäß wird eine Lyase zur Verfügung gestellt, bei der Isoleucin in dem gegenüber dem Wildtyp aus Aceobacter pasteurianus veränderten Protein ApPDC-E469G in Position Nr. 468 durch eine Aminosäure ersetzt ist, die weniger raumerfüllend als Isoleucin ist. In einer vorteilhaften Ausführungsform ist das erfindungsgemäße Enzym die Aminosäure Tryptophan in Position Nr. 543 durch eine andere Aminosäure ausgetauscht, die eine kleinere Raumerfüllung aufweist als Tryptophan. Die Erfindungsgemäßen Enzyme können zur asymmetrischen Synthese von (S)-Phenylacetylcarbinol eingesetzt werden. Dabei können Enantiomerenüberschüsse von bis zu 98% ee erreicht werden.

Device and method for detecting a specific analyte in a liquid sample and uses of said device

The invention relates to a device and a method for detecting a specific analyte in a liquid sample. The device that can be used in the method contains at least one fluid line, at least one receiving region for receiving a liquid sample, at least one enzyme region containing at least one determined enzyme and/or at least one acidification region containing at least one acid. The device also contains at least one reaction region used to form gas bubbles. The at least one fluid line is designed to transport the liquid sample from the receiving region via the enzyme region and/or the acidification region to the reaction region by means of capillary forces and/or at least one micropump. The device allows fast, simple and cost-effective detection of a specific analyte in a liquid sample, the detection being carried out with a high level of sensitivity, specificity and precision. The invention further relates to uses of the device according to the invention.

A bacterial cell factory for efficient production of ethanol from whey

The invention relates to a method for homo-ethanol production from lactose using a genetically modified lactic acid bacterium of the invention, where the cells are provided with a substrate comprising dairy waste supplemented with an amino nitrogen source (such as acid hydrolysed corn steep liquor). The invention further relates to genetically modified lactic acid bacterium and its use for homo-ethanol production from lactose in dairy waste. The lactic acid bacterium comprises both genes ( ...

Microorganism producing lactic acid and method for producing lactic acid using same

The present invention relates to a ...

USE OF BACTERIOCIN-PRODUCING ETHANOLOGENS IN BIOFUEL PRODUCTION

An ethanologen for producing biofuel from one or more carbohydrates and reducing lactate and acetate production in a biofuel manufacturing process. The ethanologen is made by introducing into the ethanologen one or more exogenous genes required for production of a bacteriocin. The resulting ethanologen reduces lactate and acetate production by contaminant lactic acid bacteria by expression of the bacteriocin during the biofuel manufacturing process. Certain resulting ethanologens ferment sugars not naturally or not preferentially utilized by Saccharomyces cerevisiae during the manufacturing process ...

METHOD FOR THE PREPARATION OF 1,3-PROPANEDIOL FROM SUCROSE

The present invention concerns a microorganism genetically modified for the bioproduction of 1,3-propanediol from sucrose,wherein the microorganism comprises: -a two-step metabolic pathway for the production of 1,3-propanediol, comprising a first step of decarboxylation of4-hydroxy-2-ketobutyrate with an enzyme having a 2-keto acid decarboxylase activity, and a second step of reduction of the obtained 3-hydroxypropionaldehyde with an enzyme having hydroxy aldehyde reductase activity,and -genes enabling the microorganism to utilize sucrose as sole carbon source. The invention also concerns a new method for the biological preparation of 1,3- propanediol by fermentation,comprising cultivating said microorganism genetically modified, wherein the culture is performed in an appropriate medium comprising a source of sucrose, and recovering the 1,3-propanediol being produced. In a preferred aspect of the invention, the source of sucrose is obtained from plant biomass.

RIFAXIMIN FOR USE IN THE TREATING OF VAGINAL INFECTIONS

젖산 분해 경로가 봉쇄된 클루이베로마이세스 막시아누스 및 이의 용도

... 본 발명은 클루이베로마이세스 막시아누스(Kluyveromycesmarxianus)에 있어서, 산화형 D-젖산 탈수소효소(oxidative D-lactate dehydrogenase)가 불활성화된 클루이베로마이세스 막시아누스 변이균주 및 상기 균주를 이용하여 젖산을 생산하는 방법에 관한 것이다.

“RECOMBINANT bacterium AND ITS USES FOR ETANOL PRODUCTION”

DEVELOPMENT OF STRAINS OF THE THERMOTOLERANT YEAST HANSENULA POLYMORPHA CAPABLE OF ALCOHOLIC FERMENTATION OF STARCH AND XYLAN BY EXPRESSION OF STARCH AND XYLAN DEGRADING ENZYMES

Genes SWA2 and GAMl from the yeast, Schwanniomyces occidentalis, encoding α- amylase and glucoamylase, respectively, were cloned and expressed in H. polymorpha. The expression was achieved by integration of the SWA2 and GAMl genes into the chromosome of H. polymorpha under operably linked to a strong constitutive promoter of the H. polymorpha - glyceraldehyde-3 -phosphate dehydrogenase gene (HpGAP. Resulting transformants acquired the ability to grow on a minimal medium containing soluble starch as a sole carbon source and can produce Ethanol at high-temperature fermentation from starch up to 10 g/L. A XYN2 gene encoding endoxylanase was obtained from the fungus Trichoderma resee, and a xlnD gene coding for β-xylosidase was obtained from the fungus Aspergillus niger. Co-expression of these genes was also achieved by integration into the H. polymorpha chromosome under control of the HpGAP promoter. The resulting transformants were capable of growth on a minimal medium supplemented with birchwood ...

Recombinant yeast for producing 2,3-butanediol including pyruvate decarboxylase derived from candida tropicolis and method for producing 2,3-butanediol using the same

Provided is a recombinant Saccharomyces cerevisiae for producing 2,3-butanediol, wherein all GPD1 and GPD2 genes involved in glycerol biosynthesis are removed and a gene encoding NADH oxidase is introduced, and wherein pyruvate decarboxylase activity is inactivated and Candida tropicalis PDC1 gene encoding Candida tropicalis pyruvate decarboxylase 1-is introduced, and wherein expression of the Candida tropicalis PDC1 gene is regulated by a GPD2 promoter.

Metabolically Enhanced Cyanobacterial Cell for the Production of Ethanol

A metabolically enhanced cyanobacterial cell for the production of ethanol is provided. The metabolically enhanced cyanobacterial cell for the production of ethanol comprises at least one recombinant gene encoding a pyruvate decarboxylase enzyme (Pdc) converting pyruvate to acetaldehyde, and at least one recombinant gene encoding a first Zn2+ dependent alcohol dehydrogenase enzyme (Adh) converting acetaldehyde to ethanol. The invention also provides a method for producing the metabolically enhanced cyanobacterium, a method for producing ethanol with the metabolically enhanced cyanobacterium, and a method for screening of alcohol dehydrogenase enzyme expressing cyanobacterial strains for the presence of NADPH-dependent native alcohol dehydrogenase enzymes.

Increased Production of Isobutanol in Yeast with Reduced Mitochondrial Amino Acid Biosynthesis

Yeast cells with reduced activity of certain enzymes involved in branched chain amino acid biosynthesis in yeast mitochondria are described. Target enzymes include threonine deaminase, isopropylmalate synthase, and optionally branched chain amino acid transaminase. 115-. (canceled)16. A recombinant yeast host cell which produces isobutanol and comprises mitochondria which is substantially devoid of branched chain amino acid transaminase activity.17. The recombinant yeast host cell of claim 16 , wherein the mitochondria is further substantially devoid of isopropylmalate synthase activity.18. The recombinant yeast host cell of claim 17 , wherein the mitochondria is further substantially devoid of threonine deaminase activity.19. The recombinant yeast cell of claim 16 , wherein the branched chain amino acid transaminase activity is defined by the enzyme classification number EC 2.6.1.42.20. The recombinant yeast cell of claim 17 , wherein the branched chain amino acid transaminase activity is defined by the enzyme classification number EC 2.6.1.42.21. The recombinant yeast cell of claim 18 , wherein the branched chain amino acid transaminase activity is defined by the enzyme classification number EC 2.6.1.42.22. The recombinant yeast host cell of claim 17 , wherein the isopropylmalate synthase activity is defined by the enzyme classification number EC 2.3.3.13.23. The recombinant yeast host cell of claim 18 , wherein the isopropylmalate synthase activity is defined by the enzyme classification number EC 2.3.3.13.24. The recombinant yeast host cell of claim 18 , wherein the threonine deaminase activity is defined by the enzyme classification number EC 4.3.1.1925. The recombinant yeast cell of claim 16 , wherein the yeast cell comprises a disruption in a BAT1 gene.26. The recombinant yeast cell of claim 17 , wherein the yeast cell comprises a disruption in a LEU4 gene.27. The recombinant yeast cell of claim 18 , wherein the yeast cell comprises a disruption in a ILV1 ...

RECOMBINANT HOST CELLS FOR THE PRODUCTION OF MALONATE

Микроорганизм, имеющий повышенную продуктивность в отношении молочной кислоты, и способ получения молочной кислоты с использованием данного микроорганизма

Fermentive production of isobutanol using highly effective ketol-acid reductoisomerase enzymes

Ketol-acid reductoisomerase enzymes have been identified that provide high effectiveness in vivo as a step in an isobutanol biosynthetic pathway in bacteria and in yeast. These KARIs are members of a clade identified through molecular phylogenetic analysis called the SLSL Clade.

INTEGRATION OF A POLYNUCLEOTIDE ENCODING A POLYPEPTIDE THAT CATALYZES PYRUVATE TO ACETOLACTATE CONVERSION

The invention relates to recombinant host cells having at least one integrated polynucleotide encoding a polypeptide that catalyzes a step in a pyruvate-utilizing biosynthetic pathway, e.g., pyruvate to acetolactate conversion. The invention also relates to methods of increasing the biosynthetic production of isobutanol, 2,3-butanediol, 2-butanol or 2-butanone using such host cells. co (D rn 4 -J Co- 0 c tOL I0 Co 0 CfOC (D >1 0) _0 Co 0)0 ...

MICROORGANISM STRAINS FOR THE PRODUCTION OF 2.3- BUTANEDIOL

A recombinant yeast having a rcduced pyruvate decarboxylase activity, in the génome of which has been inserted: - one or more nucleic acids encoding an acetolactate synthase or ALS, - one or more nucleic acids encoding an acetolactate decarboxylase or ALD, - one or more nucleic acids encoding a butancdiol dehydrogenase or BDH, and - one or more copies of a nucleic acids encoding a NADH oxidase or NOXE.

METHOD FOR PRODUCING ACETOIN

The présent invention relates to a recombinant yeast having a reduced pyruvate dccarboxylase activity, in thc génome of which has been inserted: - one or more nucleic acids encoding an acetolactate synthasc or ALS, - one or more nucleic acids encoding an acetolactate decarboxylasc or ALD, and - one or more copies of a nucleic acids encoding a NADH oxidase or NOXE.

MICROORGANISM STRAINS FOR THE PRODUCTION OF 2.3- BUTANEDIOL

A recombinant yeast having a rcduced pyruvate decarboxylase activity, in the génome of which has been inserted: - one or more nucleic acids encoding an acetolactate synthase or ALS, - one or more nucleic acids encoding an acetolactate decarboxylase or ALD, - one or more nucleic acids encoding a butancdiol dehydrogenase or BDH, and - one or more copies of a nucleic acids encoding a NADH oxidase or NOXE.

RECOMBINANT BACTERIA AND THE USES THEREOF FOR PRODUCING ETHANOL

The present invention relates to recombinant bacteria and the uses thereof, particularly for the production of ethanol. The invention also relates to methods for the production of such bacteria, as well as to nucleic acid constructs suitable for such production. The invention specifically relates to bacteria lacking a functional LDH gene and/or containing a recombinant nucleic acid encoding a PDC and ADH. The bacteria of this invention may be produced from any stress-resistant bacteria.

FERMENTIVE PRODUCTION OF ISOBUTANOL USING HIGHLY EFFECTIVE KETOL-ACID REDUCTOISOMERASE ENZYMES

Ketol-acid reductoisomerase enzymes have been identified that provide high effectiveness in vivo as a step in an isobutanol biosynthetic pathway in bacteria and in yeast. These KARIs are members of a clade identified through molecular phylogenetic analysis called the SLSL Clade.

INTEGRATION OF A POLYNUCLEOTIDE ENCODING A POLYPEPTIDE THAT CATALYZES PYRUVATE TO ACETOLACTATE CONVERSION

두날리엘라 살리나 유래의 키메릭 탄산무수화효소 및 이의 용도

... 본 발명은 두날리엘라 살리나 유래의 키메릭 탄산무수화효소 및 이의 용도에 관한 것으로, 두날리엘라 살리나 유래의 키메릭 탄산무수화효소는 에스테르 분해 활성 및 이산화탄소 수화 활성이 우수하여 산업용 이산화탄소 포집 및 고정화 공정 개발에 사용할 수 있고, 바이카보네이트 화합물 제조에 사용할 수 있을 뿐만 아니라, 유용 대사 산물 합성 시 촉매로 사용되어 이의 합성 수율을 높일 수 있다.

Genetically modified yeast species and fermentation processes using genetically modified yeast

Yeast cells are transformed with an exogenous xylose isomerase gene. Additional genetic modifications enhance the ability of the transformed cells to ferment xylose to ethanol or other desired fermentation products. Those modifications', include deletion of non-specific or specific aldose reductase gene(s), deletion of xylitol dehydrogenase gene(s) and/or overexpression of xylulokinase.

3-hydroxypropionic acid production by recombinant yeasts expressing an insect aspartate 1-decarboxylase

Provided herein are recombinant yeast cells having an active 3-Hydroxypropionic Acid (3-HP) pathway and further comprising a heterologous polynucleotide encoding an aspartate 1-decarboxylase (ADC) of the Class Insecta, Bivalvia, Branchioporia, Gastropoda, or Leptocardii. Also described are methods of using the recombinant yeast cells to produce 3-HP and acrylic acid.

ENZYMATIC METHOD FOR PRODUCING 2-HYDROXY-4-METHYLMERCAPTOBUTANOIC ACID (MHA)

СПОСОБ ПОЛУЧЕНИЯ АЦЕТОИНА

Настоящее изобретение относится к рекомбинантным дрожжам, имеющим пониженную активность пируватдекарбоксилазы, в геном которых вставлена:- одна или более нуклеиновых кислот, кодирующих ацетолактатсинтазу или ALS,- одна или более нуклеиновых кислот, кодирующих ацетолактатдекарбоксилазу или ALD, и- одна или более копий нуклеиновых кислот, кодирующих NADH-оксидазу или NOXE.

Recombinant bacteria and the uses thereof for producing ethanol

The present invention relates to recombinant bacteria and the uses thereof, particularly for the production of ethanol. The invention also relates to methods for the production of such bacteria, as well as to nucleic acid constructs suitable for such production. The invention specifically relates to bacteria lacking a functional LDH gene and/or containing a recombinant nucleic acid encoding a PDC and ADH. The bacteria of this invention may be produced from any stress-resistant bacteria.

Recombinant host cells comprising phosphoketolases

The present invention is related to recombinant host cells comprising: (i) at least one deletion, mutation, and/or substitution in an endogenous gene encoding a polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA; and (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. The present invention is also related to recombinant host cells further comprising (iii) a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.

AN ELECTRON CONSUMING ETHANOL PRODUCTION PATHWAY TO DISPLACE GLYCEROL FORMATION IN S. CEREVISIAE

The present invention provides for a mechanism to completely replace the electron accepting function of glycerol formation with an alternative pathway to ethanol formation, thereby reducing glycerol production and increasing ethanol production. In some embodiments, the invention provides for a recombinant microorganism comprising a down-regulation in one or more native enzymes in the glycerol-production pathway. In some embodiments, the invention provides for a recombinant microorganism comprising an up-regulation in one or more enzymes in the ethanol-production pathway.

Integration of a polynucleotide encoding a polypeptide that catalyzes pyruvate to acetolactate conversion

The invention relates to recombinant host cells having at least one integrated polynucleotide encoding a polypeptide that catalyzes a step in a pyruvate-utilizing biosynthetic pathway, e.g., pyruvate to acetolactate conversion. The invention also relates to methods of increasing the biosynthetic production of isobutanol, 2,3-butanediol, 2-butanol or 2-butanone using such host cells.

NOVEL KLUYVEROMYCES MARXIANUS MJ1 AND USE THEREOF

The present invention relates to a novel Kluyveromyces marxianus MJ1 strain, which exhibits thermostability and acid tolerance. The present invention further relates to a composition which is used for producing lactic acid and contains both the strain and a culture material thereof, and to a lactic acid production method including a step of culturing the strain. COPYRIGHT KIPO 2016 (AA) Adjusted to pH 3.0 (BB) Adjusted to pH 3.5 ...

METHOD FOR PRODUCING COENZYME OF NICOTINAMIDE-DEPENDENT OXIDOREDUCTASE, AND USE THEREOF

Disclosed in the present invention are a coenzyme of nicotinamide-dependent oxidoreductase and a method for producing the coenzyme of nicotinamide-dependent oxidoreductase. According to the method of the present invention, the production efficiency of the coenzyme of nicotinamide-dependent oxidoreductase is maximized, thereby being able to be helpfully used in various oxidoreductase reactions such as the improvement of ethanol production efficiency by combining the oxidoreductase reaction to an ethanol production circuit. COPYRIGHT KIPO 2016 ...

METHODS FOR PRODUCTION OF ISOBUTANOL THROUGH NUTRIENT STRESS AND GENETICALLY MODIFIED MICROORGANISMS THEREOF

The present disclosure relates to an architecture of energy redistribution that can sustain the increased formation of cofactors like NADH/NADPH and key metabolites like pyruvate that are implicated in the production of isobutanol through biotransformation. Genetically modified microorganisms comprising altered genes are disclosed wherein said alteration optionally along with subjecting the genetically modified microorganism to nutrient stress induces redistribution of energy ultimately resulting in maximum production of isobutanol.

Microorganisms and methods for production of specific length fatty alcohols and related compounds

The invention provides non-naturally occurring microbial organisms containing a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length. Also provided are non-naturally occurring microbial organisms having a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms further include an acetyl-CoA pathway. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde or a fatty acid.

GENETICALLY MODIFIED YEAST SPECIES, AND FERMENTATION PROCESSES USING GENETICALLY MODIFIED YEAST

Yeastcells are transformed with an exogenous xylose isomerase gene. Additional genetic modifications enhance the ability of the transformed cells to ferment xylose to ethanol or other desired fermentation products. Those modifications include deletion of non-specific or specific aldose reductase gene(s), deletion of xylitol dehydrogenase gene(s) and/or overexpression of xylulokinase.

Genetically Modified Yeast Species, and Fermentation Processes Using Genetically Modified Yeast

Yeast cells are transformed with an exogenous xylose isomerase gene. Additional genetic modifications enhance the ability of the transformed cells to ferment xylose to ethanol or other desired fermentation products. Those modifications include deletion of non-specific or specific aldose reductase gene(s), deletion of xylitol dehydrogenase gene(s) and/or overexpression of xylulokinase. 145-. (canceled)46. A fermentation process in which a genetically modified yeast cell having a genome and a functional , exogenous xylose isomerase gene which encodes a xylose isomerase enzyme that is at least 90% homologous to SEQ. ID. NO. 59 , wherein the exogenous xylose isomerase gene is operatively linked to promoter and terminator sequences that are functional in the yeast cell , and the modified yeast cell further has a deletion or disruption of a native gene that produces an enzyme that catalyzes the conversion of xylose to xylitol is cultured under fermentation conditions in a fermentation broth that includes a pentose sugar , wherein the fermentation process includes a growth phase and a production phase , and during the production phase the concentration of cells in the fermentation broth is in the range of 3 to 10 g dry cells/liter of fermentation broth.47. A fermentation process of wherein ethanol is produced as a major fermentation product.48. A fermentation process of wherein lactate is produced as a major fermentation product.49. The fermentation process of in which the pentose sugar includes xylose.50. The fermentation process of in which the fermentation broth further includes a hexose sugar.51. The fermentation process of which is an anaerobic fermentation.52108-. (canceled)109. The fermentation process of wherein the final concentration of the fermentation product is at least 14 g/L. This application claims benefit of U.S. Provisional Application No. 60/467,727, filed May 2, 2003.This invention was made under contract no. DE-FC07-021D14349 with the United States ...

USE OF BACTERIOCIN-PRODUCING ETHANOLOGENS IN BIOFUEL PRODUCTION

An ethanologen for producing biofuel from one or more carbohydrates and reducing lactate and acetate production in a biofuel manufacturing process. The ethanologen is made by introducing into the ethanologen one or more exogenous genes required for production of a bacteriocin. The resulting ethanologen reduces lactate and acetate production by contaminant lactic acid bacteria by expression of the bacteriocin during the biofuel manufacturing process. Certain resulting ethanologens ferment sugars not naturally or not preferentially utilized by during the manufacturing process 122-. (canceled)23. An ethanologen comprising(a) one or more inactivated endogenous genes encoding a mannitol dehydrogenase;(b) one or more inactivated endogenous genes encoding a lactate dehydrogenase; and(c) one or more exogenous genes encoding a pyruvate decarboxylase and one or more exogenous genes encoding an alcohol dehydrogenase;wherein the ethanologen is an engineered lactic acid bacterium; andwhereby the engineered lactic acid bacterium produces more biofuel than a wild-type lactic acid bacterium having the same genetic background in a biofuel manufacturing process24. The ethanologen of claim 23 , wherein the one or more inactivated endogenous genes encoding a mannitol dehydrogenase comprises a deletion in a mannitol dehydrogenase 1 and/or a mannitol dehydrogenase 2 gene.25. The ethanologen of claim 23 , wherein the one or more inactivated endogenous genes encoding a lactate dehydrogenase comprises a deletion in a lactate dehydrogenase 1 (LDH1) claim 23 , lactate dehydrogenase 2 (LDH2) claim 23 , lactate dehydrogenase 3 (LDH3) claim 23 , and/or lactate dehydrogenase 4 (LDH4) gene.26. The ethanologen of claim 23 , wherein the ethanologen is capable of fermenting sugars not naturally or not preferentially fermented by a main fermenting microbe present in the biofuel manufacturing process.27Saccharomyces cerevisiae.. The ethanologen of claim 26 , wherein the main fermenting microbe ...

Микроорганизм, продуцирующий молочную кислоту, и способ продуцирования молочной кислоты с его использованием

Группа изобретений относится к микроорганизму рода Saccharomyces, обладающему способностью продуцировать молочную кислоту по сравнению с немодифицированным микроорганизмом, и к способу продуцирования молочной кислоты с его использованием. Микроорганизм рода Saccharomyces модифицирован так, что у него инактивирована активность пируватдекарбоксилазы (PDC) по сравнению с ее эндогенной активностью, введена активность АТР-цитратлиазы (ACL) и усилен путь биосинтеза пирувата по сравнению с эндогенным путем его биосинтеза. Способ получения молочной кислоты включает культивирование указанного микроорганизма рода Saccharomyces в среде и выделение молочной кислоты из культивируемого микроорганизма или из среды. Группа изобретений обеспечивает увеличение количества продуцируемой молочной кислоты. 2 н. и 10 з.п. ф-лы, 2 ил., 13 табл., 10 пр.

Protein manipulation

Production of glucan polymers from alternate sucrose sources

Reaction solutions are disclosed herein comprising water, incompletely refined sucrose, and a glucosyltransferase enzyme that synthesizes insoluble poly alpha-1,3-glucan having at least 50% alpha-1,3 glycosidic linkages and a weight average degree of polymerization (DP ...

INTEGRATION OF A POLYNUCLEOTIDE ENCODING A POLYPEPTIDE THAT CATALYZES PYRUVATE TO ACETOLACTATE CONVERSION

The invention relates to recombinant host cells having at least one integrated polynucleotide encoding a polypeptide that catalyzes a step in a pyruvate-utilizing biosynthetic pathway, e.g., pyruvate to acetolactate conversion. The invention also relates to methods of increasing the biosynthetic production of isobutanol, 2,3-butanediol, 2-butanol or 2-butanone using such host cells. co (D rn 4 -J Co- 0 c tOL I0 Co 0 CfOC (D >1 0) _0 Co 0)0 ...

Method for producing acetoin

The présent invention relates to a recombinant yeast having a reduced pyruvate dccarboxylase activity, in thc génome of which has been inserted: - one or more nucleic acids encoding an acetolactate synthasc or ALS, - one or more nucleic acids encoding an acetolactate decarboxylasc or ALD, and - one or more copies of a nucleic acids encoding a NADH oxidase or NOXE.

METHOD FOR PRODUCING ACETOIN

The présent invention relates to a recombinant yeast having a reduced pyruvate dccarboxylase activity, in thc génome of which has been inserted: - one or more nucleic acids encoding an acetolactate synthasc or ALS, - one or more nucleic acids encoding an acetolactate decarboxylasc or ALD, and - one or more copies of a nucleic acids encoding a NADH oxidase or NOXE.

CARBONIC ANHYDRASE-IMMOBILIZED SILK HYDROGEL AND CONVERSION OR FIXATION OF CARBON DIOXIDE USING SAME

The present invention relates to carbonic anhydrase-immobilized silk hydrogel and the conversion or the fixation of carbon dioxide using the same. According to the present invention, the carbonic anhydrase-immobilized silk hydrogel or a composition comprising the same is dual crosslinked by the photocrosslinking and alcohol treatment, thereby being eco-friendly and economic, having excellent thermal and storage stability, and being used in a repeated manner. The carbonic anhydrase-immobilized silk hydrogel or the composition comprising the same has significantly excellent effects of enzymatic activities, thereby removing, converting, or fixing carbon dioxide. COPYRIGHT KIPO 2016 (AA) m-ngCA-silk hydrogel (BB) ngCA-silk hydrogel ...

“CEPA H. POLYMORPHA, PROCESS FOR THE MANUFACTURE OF RECOMBINANT ETANOL AND ACID NUCLÉICO”

USE OF BACTERIOCIN-PRODUCING ETHANOLOGENS IN BIOFUEL PRODUCTION

METHOD FOR THE PREPARATION OF 1,3-PROPANEDIOL FROM SUCROSE

Metabolically enhanced cyanobacterial cell for the production of ethanol

A metabolically enhanced cyanobacterial cell for the production of ethanol is provided. The metabolically enhanced cyanobacterial cell for the production of ethanol comprises at least one recombinant gene encoding a pyruvate decarboxylase enzyme (Pdc) converting pyruvate to acetaldehyde, and at least one recombinant gene encoding a first Zn2+ dependent alcohol dehydrogenase enzyme (Adh) converting acetaldehyde to ethanol. The invention also provides a method for producing the metabolically enhanced cyanobacterium, a method for producing ethanol with the metabolically enhanced cyanobacterium, and a method for screening of alcohol dehydrogenase enzyme expressing cyanobacterial strains for the presence of NADPH-dependent native alcohol dehydrogenase enzymes.

Ethanol production by recombinant hosts

Compositions and methods for 3-hydroxypropionate bio-production from biomass

Methods of obtaining mutant nucleic acid sequences that demonstrate elevated oxaloacetate -decarboxylase activity are provided. Compositions, such as genetically modified microorganisms that comprise such mutant nucleic acid sequences, are described, as are methods to obtain the same.

METHODS AND COMPOSITIONS FOR GENETIC ENGINEERING OF CYANOBACTERIA TO PRODUCE ETHANOL

Штамм дрожжей Schizosaccharomyces pombe, продуцирующий L-молочную кислоту, содержащий в составе хромосомы гены трех различных гетерологичных лактатдегидрогеназ

Изобретение относится к биотехнологии. Предложен рекомбинантный штамм Schizosaccharomyces pombe ВКПМ Y-4822, являющийся продуцентом L-молочной кислоты. Штамм имеет делетированный ген пируватдекарбоксилазы PDC1 и несет в составе хромосомной ДНК гены лактатдегидрогеназ из Lactobacillus pentosus, Lactobacillus acidophilus и Lactobacillus helveticus. Штамм продуцирует L-молочную кислоту в количестве 125 г/л за 72 ч культивирования в ферментере в аэробных условиях. Изобретение позволяет расширить арсенал рекомбинантных микроорганизмов, продуцирующих L-молочную кислоту. 2 ил., 3 пр.

Genetically modified yeast species and fermentation processes using genetically modified yeast

Integration of a polynucleotide encoding a polypeptide that catalyzes pyruvate to acetolactate conversion

The invention relates to recombinant host cells having at least one integrated polynucleotide encoding a polypeptide that catalyzes a step in a pyruvate-utilizing biosynthetic pathway, ...

BACTERIA ENGINEERED TO TREAT DISORDERS INVOLVING THE CATABOLISM OF A BRANCHED CHAIN AMINO ACID

The present disclosure provides recombinant bacterial cells that have been engineered with genetic circuitry which allow the recombinant bacterial cells to sense a patient's internal environment and respond by turning an engineered metabolic pathway on or off. When turned on, the recombinant bacterial cells complete all of the steps in a metabolic pathway to achieve a therapeutic effect in a host subject. These recombinant bacterial cells are designed to drive therapeutic effects throughout the body of a host from a point of origin of the microbiome. Specifically, the present disclosure provides recombinant bacterial cells comprising a heterologous gene encoding a branched chain amino acid catabolism enzyme. The disclosure further provides pharmaceutical compositions comprising the recombinant bacteria, and methods for treating disorders involving the catabolism of branched chain amino acids using the pharmaceutical compositions disclosed herein.

YEAST ORGANISM PRODUCING ISOBUTANOL AT A HIGH YIELD

There is disclosed a method of producing isobutanol. In an embodiment, the method includes providing a microorganism transformed with an isobutanol producing pathway containing at least one exogenous gene. The microorganism is selected to produce isobutanol from a carbon source at a yield of at least 10 percent theoretical. The method includes cultivating the microorganism in a culture medium containing a feedstock providing the carbon source, until isobutanol is produced. The method includes recovering the isobutanol. In one embodiment, the microorganism is a yeast with a Crabtree-negative phenotype. In another embodiment, the microorganism is a yeast microorganism with a Crabtree-positive phenotype. There is disclosed a microorganism for producing isobutanol. In an embodiment, the microorganism includes an isobutanol producing pathway containing at least one exogenous gene, and is selected to produce a recoverable quantity of isobutanol from a carbon source at a yield of at least 10 percent ...

YEAST ORGANISM PRODUCING ISOBUTANOL AT A HIGH YIELD

There is disclosed a method of producing isobutanol. Sn an embodiment, the method includes providing a microorganism transformed with an isobutanoi producing pathway containing at least one exogenous gene. The microorganism is selected to produce isobutanol from a carbon source at a yield of at least 10 percent theoretical. The method includes cultivating the microorganism in a culture medium containing a feedstock providing the carbon source, until isobutanol is produced. The method includes recovering the isobutanol. In one embodiment, the microorganism is a yeast with a Crabtree-negative phenotype. In another embodiment, the microorganism is a yeast microorganism with a Crabtree-positive phenotype. There is disclosed a microorganism for producing isobutanol. In an embodiment, the microorganism includes an isobutanol producing pathway containing at least one exogenous gene, and is selected to produce a recoverable quantity of isobutanol from a carbon source at a yield of at least 10 percent ...

TRANSFORMANT FOR ENHANCING BIOETHANOL PRODUCTION, AND METHOD FOR PRODUCING ETHANOL BY USING SAID STRAIN

The present invention relates to a transformant for inhibiting glycerol production through deletion of glycerol producing genes of Saccharomyces cerevisiae modified so as to use glycerol as a fermentation source, or enhancing bioethanol production through overexpression of TATA-binding proteins, SPT3 and SPT15, and a method for producing ethanol by using the transformant.

TRANSFORMANT FOR ENHANCING BIOETHANOL PRODUCTION, AND METHOD FOR PRODUCING ETHANOL BY USING SAID STRAIN

The present invention relates to a transformant for inhibiting glycerol production through deletion of glycerol producing genes of Saccharomyces cerevisiae modified so as to use glycerol as a fermentation source, or enhancing bioethanol production through overexpression of TATA-binding proteins, SPT3 and SPT15, and a method for producing ethanol by using the transformant.

Butanol strain improvement with integration of a polynucleotide encoding a polypeptide that catalyzes pyruvate to acetolactate conversion

The invention relates to recombinant host cells having at least one integrated polynucleotide encoding a polypeptide that catalyzes a step in a pyruvate-utilizing biosynthetic pathway, e.g., pyruvate to acetolactate conversion. The invention also relates to methods of increasing the biosynthetic production of isobutanol, 2,3-butanediol, 2-butanol or 2-butanone using such host cells.

Butanol tolerance in microorganisms

Provided herein are recombinant yeast host cells and methods for their use for production of fermentation products from a pyruvate utilizing pathway. Yeast host cells provided herein comprise reduced pyruvate decarboxylase activity and modified adenylate cyclase activity. In embodiments, yeast host cells provided herein comprise resistance to butanol and increased biomass production.

Electron consuming ethanol production pathway to displace glycerol formation in S. cerevisiae

The present invention provides for a mechanism to completely replace the electron accepting function of glycerol formation with an alternative pathway to ethanol formation, thereby reducing glycerol production and increasing ethanol production. In some embodiments, the invention provides for a recombinant microorganism comprising a down-regulation in one or more native enzymes in the glycerol-production pathway. In some embodiments, the invention provides for a recombinant microorganism comprising an up-regulation in one or more enzymes in the ethanol-production pathway.

Modified microorganisms and methods for production of useful products

Recombinant host cells comprising phosphoketolases

The present invention is related to recombinant host cells comprising: (i) at least one deletion, mutation, and/or substitution in an endogenous gene encoding a polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA; and (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. The present invention is also related to recombinant host cells further comprising (iii) a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.

Production of fermentation products

The invention relates to the development of microorganisms capable of producing fermentation products via an engineered pathway in the microorganisms. The invention also relates to microorganisms with improved cell viability and methods to improve cell viability and cell productivity of a microorganism.

Gene therapy for the treatment of aldehyde dehydrogenase deficiency

A vector comprising a promoter operably linked to a nucleic acid sequence encoding human aldehyde dehydrogenase, as well as a composition comprising the vector and method of using the vector to treat aldehyde dehydrogenase deficiency, or to prevent or treat a disease characterized by aldehyde dehydrogenase deficiency.

Method for producing acetoin

The présent invention relates to a recombinant yeast having a reduced pyruvate dccarboxylase activity, in thc génome of which has been inserted: - one or more nucleic acids encoding an acetolactate synthasc or ALS, - one or more nucleic acids encoding an acetolactate decarboxylasc or ALD, and - one or more copies of a nucleic acids encoding a NADH oxidase or NOXE.

Processes for ethanol production

INTEGRATION OF A POLYNUCLEOTIDE ENCODING A POLYPEPTIDE THAT CATALYZES PYRUVATE TO ACETOLACTATE CONVERSION

POLYPEPTIDES WITH KETOL-ACID REDUCTOISOMERASE ACTIVITY

Polypeptides having ketol-acid reductoisomerase activity are provided. Also disclosed are recombinant host cells comprising isobutanol biosynthetic pathways employing such polypeptides. Methods for producing isobutanol employing host cells comprising the polypeptides having ketol-acid reductoisomerase activity are also disclosed.

MICROORGANISMS AND METHODS FOR PRODUCTION OF SPECIFIC LENGTH FATTY ALCOHOLS AND RELATED COMPOUNDS

The invention provides non-naturally occurring microbial organisms containing a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length. Also provided are non-naturally occurring microbial organisms having a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms further include an acetyl-CoA pathway. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde or a fatty acid.

MODIFIED MICROORGANISMS AND METHODS FOR PRODUCTION OF USEFUL PRODUCTS

The invention provides non-naturally occurring microbial organisms and related methods, processes and materials wherein the microbial organisms includes a genetic modification which enhances production of 3-hydroxybutanal or a downstream product of 3- hydroxybutanal such as 1,3-butanediol from endogenous central metabolic intermediates such as acetyl CoA or pyruvate which are converted to acetaldehyde, whereby two molecules of acetaldehyde are condensed to form said 3-hydroxybutanal using an aldolase capable of accepting acetaldehyde as both the acceptor and donor in an aldol condensation. The aldolase may be a deoxyribose phosphate aldolase type enzyme, and is typically introduced into the organisms. The invention provides for energetically favorable pathways for production of 3-hydroxybutanal or downstream products thereof.

Microorganisms and methods for production of specific length fatty alcohols and related compounds

The invention provides non-naturally occurring microbial organisms containing a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length. Also provided are non-naturally occurring microbial organisms having a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms further include an acetyl-CoA pathway. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde or a fatty acid.

RECOMBINANT HOST CELLS COMPRISING PHOSPHOKETOLASES

The present invention is related to recombinant host cells comprising: (i) at least one deletion, mutation, and/or substitution in an endogenous gene encoding a polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA; and (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. The present invention is also related to recombinant host cells further comprising (iii) a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.

ELECTRON CONSUMING ETHANOL PRODUCTION PATHWAY TO DISPLACE GLYCEROL FORMATION IN S. CEREVISIAE

The present invention provides for a mechanism to completely replace the electron accepting function of glycerol formation with an alternative pathway to ethanol formation, thereby reducing glycerol production and increasing ethanol production. In some embodiments, the invention provides for a recombinant microorganism comprising a down-regulation in one or more native enzymes in the glycerol-production pathway. In some embodiments, the invention provides for a recombinant microorganism comprising an up-regulation in one or more enzymes in the ethanol-production pathway.

Engineered cyanobacterium and its application for producing acetate

The present invention provides an engineered cyanobacterium, comprising at least one plasmid selected from three novel pathways to produce acetate, which can convert atmospheric carbon dioxide as a raw material into acetate. The present invention also constructs the expression plasmid for three different transporters specific to acetate to be expressed in cyanobacteria, which comprises putative ABC transporter (AatA), succinate/acetate: proton symporter (SatP) and acetate/glycolate: cation symporter (ActP). Therefore, the engineered cyanobacteria of the present invention can produce 0.58 mg/L to 3.54 mg/L of acetate per hour.

Electron consuming ethanol production pathway to displace glycerol formation in S. cerevisiae

The present invention provides for a mechanism to completely replace the electron accepting function of glycerol formation with an alternative pathway to ethanol formation, thereby reducing glycerol production and increasing ethanol production. In some embodiments, the invention provides for a recombinant microorganism comprising a down-regulation in one or more native enzymes in the glycerol-production pathway. In some embodiments, the invention provides for a recombinant microorganism comprising an up-regulation in one or more enzymes in the ethanol-production pathway.

Xylose isomerase-modified yeast strains and methods for bioproduct production

The present invention relates to a genetically engineered yeast (e.g., Crabtree negative) that express a heterologous xylose isomerase. In a fermentation method the engineered yeast are capable of producing a bioproduct, such as ethanol, in a fermentation medium that includes xylose. Desirable bioproduct titers can be achieved when materials such as acetate are present in the fermentation medium.

Acetate supplemention of medium for butanologens

The invention relates to the fields of industrial microbiology and alcohol production. More specifically, the invention relates to improved production of butanol isomers by recombinant microorganisms containing an engineered butanol pathway and disrupted activity of the genes in pathways for the production of by-products during the fermentation when the microorganisms are grown in a fermentation medium containing acetate. In embodiments, recombinant microorganisms have an increased growth rate in a fermentation medium containing acetate as a C2 supplement.

Fermentive production of isobutanol using highly effective ketol-acid reductoisomerase enzymes

Ketol-acid reductoisomerase enzymes have been identified that provide high effectiveness in vivo as a step in an isobutanol biosynthetic pathway in bacteria and in yeast. These KARIs are members of a clade identified through molecular phylogenetic analysis called the SLSL Clade.

Production of glucan polymers from alternate sucrose sources

Reaction solutions are disclosed herein comprising water, incompletely refined sucrose, and a glucosyltransferase enzyme that synthesizes insoluble poly alpha-1,3-glucan having at least 50% alpha-1,3 glycosidic linkages and a weight average degree of polymerization (DP ...

METHOD FOR PRODUCING LACTIC ACID

The present invention provides a method for producing lactic acid in a recombinant yeast cell culture using glucose as carbon source comprising a first, seed fermentation stage to produce biomass wherein the yeast is cultivated in a culture medium at a p H of 5 to 7, followed by a second, a production fermentation stage with biomass from the seed fermentation to produce lactic acid, wherein the yeast is cultivated in a culture medium at low p H using a yeast strain that is engineered to have lactate dehydrogenase (LDH) activity and optionally has decreased or knocked- out pyruvate decarboxylase (PDC) activity.

수크로스로부터의 1,3-프로판디올의 제조 방법

... 본 발명은 - 4-히드록시-2-케토부티레이트를 2-케토산 데카르복실라제 활성을 갖는 효소로 탈카르복실화시키는 제1 단계, 및 수득한 3-히드록시프로피온알데히드를 히드록시 알데히드 리덕타제 활성을 갖는 효소로 환원시키는 제2 단계를 포함하는, 1,3-프로판디올의 생산을 위한 2-단계 대사 경로, 및 - 미생물이 단일 탄소원으로 수크로스를 이용할 수 있도록 하는 유전자 를 포함하는, 수크로스로부터의 1,3-프로판디올의 생물생산을 위해 유전자 변형된 미생물에 관한 것이다. 본 발명은 또한 상기 유전자 변형된 미생물을 배양하는 것 (여기서 배양은 수크로스의 공급원을 포함하는 적절한 배지에서 수행함), 및 생산되는 1,3-프로판디올을 회수하는 것을 포함하는, 발효에 의한 1,3-프로판디올의 생물학적 제조를 위한 새로운 방법에 관한 것이다. 본 발명의 바람직한 측면에서, 수크로스의 공급원은 식물 바이오매스로부터 수득된다.

Yeast organism producing isobutanol at a high yield

The present invention provides recombinant microorganisms comprising an isobutanol producing metabolic pathway and methods of using said recombinant microorganisms to produce isobutanol. In various aspects of the invention, the recombinant microorganisms may comprise a modification resulting in the reduction of pyruvate decarboxylase and/or glycerol-3-phosphate dehydrogenase activity. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.

Acetate supplemention of medium for butanologens

The invention relates to the fields of industrial microbiology and alcohol production. More specifically, the invention relates to improved production of butanol isomers by recombinant microorganisms containing an engineered butanol pathway and disrupted activity of the genes in pathways for the production of by-products during the fermentation when the microorganisms are grown in a fermentation medium containing acetate. In embodiments, recombinant microorganisms have an increased growth rate in a fermentation medium containing acetate as a C2 supplement.

Construction of a lactobacillus casei ethanologen

An engineered bacterium for producing ethanol from one or more carbohydrates is disclosed. The bacterium can be made by (a) inactivating within a Lactobacillus casei bacterium one or more endogenous genes encoding a lactate dehydrogenase; or (b) introducing into a Lactobacillus casei bacterium one or more exogenous genes encoding a pyruvate decarboxylase and one or more exogenous genes encoding an alcohol dehydrogenase II; or (c) performing both steps (a) and (b). The resulting engineered bacterium produces significantly more ethanol than the wild-type Lactobacillus casei bacterium, and can be used in producing ethanol from a substrate such as biomass that includes carbohydrates.

YEAST HAVING IMPROVED PRODUCTIVITY AND METHOD OF PRODUCING PRODUCT

A recombinant yeast cell capable of consuming glucose at an increased rate, and a method of efficiently producing glycolysis-derived products using the recombinant yeast cell. 1. A recombinant yeast cell having increased activity of at least one of GCR1 and GCR2 , wherein the recombinant yeast cell comprises a genetic modification that increases activity of at least one of GCR1 and GCR2 , in comparison with a yeast cell of the same type that does not comprise the genetic modification that increases activity of at least one of GCR1 and GCR2.2. The recombinant yeast cell of claim 1 , wherein the yeast cell is capable of consuming glucose at an increased glucose consumption rate in comparison with a yeast cell of the same type that does not comprise the genetic modification that increases activity of at least one of GCR1 and GCR2.3. The recombinant yeast cell of claim 1 , wherein the yeast cell has an increased productivity of a glycolysis intermediate or glycolysis intermediate-derived material in comparison with a yeast cell of the same type that does not comprise the genetic modification that increases activity of at least one of GCR1 and GCR2.4. The recombinant yeast cell of claim 3 , wherein the glycolysis intermediate comprises dihydroxyacetone phosphate (DHAP) claim 3 , glyceraldehyde 3-phosphate (GAP) claim 3 , or pyruvate claim 3 , and wherein the glycolysis intermediates-derived material comprises glyceol-3-phosphate (G3P) claim 3 , glycerol claim 3 , acetyl-CoA claim 3 , ethanol claim 3 , acetic acid claim 3 , lactate claim 3 , citric acid claim 3 , itaconic acid claim 3 , isocitric acid claim 3 , oxalosuccinic acid claim 3 , α-ketoglutaric acid claim 3 , succinic acid claim 3 , succinyl-CoA claim 3 , fumaric acid claim 3 , maleic acid claim 3 , oxaloacetic acid claim 3 , 1 claim 3 ,3-butanediol (1 claim 3 ,3-BDO) claim 3 , 1 claim 3 ,4-butanediol (1 claim 3 ,4-BDO) claim 3 , butanol claim 3 , isobutanol claim 3 , or a combination thereof.5. The recombinant ...

MICROORGANISMS THAT CO-CONSUME GLUCOSE WITH NON-GLUCOSE CARBOHYDRATES AND METHODS OF USE

Microorganisms that co-consume glucose with non-glucose carbohydrates, such as xylose, and methods of using same. The microorganisms comprise modifications that reduce or ablate the activity of a phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS) protein or modifications that reduce or ablate the activity of a phosphoglucose isomerase and a GntR. The PTS protein may be selected from an enzyme I (EI), an HPr, an FPr, and an enzyme II(EII). Additional modifications include reduction or ablation of the activity of a pyruvate formate lyase, a lactate dehydrogenase, and a fumarate reductase and inclusion of recombinant pyruvate decarboxylase and alcohol dehydrogenase genes. The microorganisms are particularly suited to co-consuming glucose and xylose in media containing these substrates and producing ethanol therefrom. 1. A recombinant microorganism comprising:{'sup': Glc', 'Glc, 'modifications that reduce or ablate the activity of a phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS) protein selected from the group consisting of an enzyme I (EI), an HPr, an FPr, and an enzyme II(EII); or'}a first modification that reduces or ablates the activity of a phosphoglucose isomerase and a second modification selected from the group consisting of a modification that reduces or ablates the activity of a GntR, a modification that introduces a recombinant phosphogluconate dehydratase gene, a modification that introduces a recombinant 2-keto-4-hydroxyglutarate aldolase gene, a modification that introduces a recombinant 2-keto-3-deoxy-6-phosphogluconate aldolase gene, and a modification that introduces a recombinant oxaloacetate decarboxylase gene.2. The recombinant microorganism of wherein the microorganism is a bacterium.3E. coliE. coli. The recombinant microorganism of comprising a modification that reduces or ablates the activity of HPr of or an ortholog thereof and FPr of or an ortholog thereof.4. The recombinant microorganism of further ...

Discovery of enzymes from the alpha-keto acid decarboxylase family

2-ketoacid decarboxylase enzymes, compositions encoding for 2 ketoacid decarboxylase enzymes, and host cells comprising such enzymes or compositions are provided

METHOD FOR PRODUCING 1,3-PROPANEDIOL USING MICROORGANISM VARIANT WITH DELETION OF 2,3-BUTANEDIOL SYNTHETIC GENE

The present invention relates to a method for producing 1,3-propanediol using a mutant microorganism lacking a 2,3-butanediol synthetic gene, and more particularly to a mutant microorganism wherein a gene encoding lactate dehydrogenase and a gene encoding an enzyme which is involved in 2,3-butanediol synthesis are deleted in a microorganism having the ability to produce 1,3-propanediol from glycerol and wherein a gene encoding pyruvate decarboxylase and a gene encoding aldehyde dehydrogenase are introduced or amplified, and to a method of promoting the production of 1,3-propanediol while inhibiting the production of 2,3-butanediol by using the mutant microorganism. The use of the glycerol-fermenting mutant microorganism according to the present invention can significantly increase the production of 1,3-propanediol while minimizing the production of 2,3-butanediol. 1. A mutant microorganism wherein a gene encoding lactate dehydrogenase and a gene encoding an enzyme which is involved in 2 ,3-butanediol synthesis are deleted in a microorganism having the ability to produce 1 ,3-propanediol from glycerol and wherein a gene encoding pyruvate decarboxylase and a gene encoding aldehyde dehydrogenase are introduced or amplified.2. The mutant microorganism of claim 1 , wherein the enzyme which is involved in 2 claim 1 ,3-butanediol synthesis is an acetolactate synthase.3. The mutant microorganism of claim 1 , wherein the gene encoding pyruvate decarboxylase is pdc derived from a stain claim 1 , which has pyruvate decarboxylase activity.4. The mutant microorganism of claim 1 , wherein the gene encoding aldehyde dehydrogenase is aldB derived from a strain claim 1 , which has pyruvate decarboxylase activity.5. The mutant microorganism of claim 1 , which is Klebsiella pneumonia.6. A method for producing 1 claim 1 ,3-propanediol claim 1 , comprising the steps of:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) culturing the mutant microorganism of in a glycerol-containing ...

MICROORGANISMS AND PROCESSES FOR LACTIC ACID PRODUCTION

Improved yeast strains and fermentation process for producing D-lactic acid and L-lactic acid are disclosed. The improvement lead to higher titer, higher yield, shorter time, lower pH, and higher average specific productivity. 1. A yeast strain genetically engineered to produce lactic acid comprising a chromosomally integrated gene that encodes an exogenous lactate dehydrogenase , wherein said yeast strain produces lactic acid in a fermentation production medium with an average specific productivity of at least 1.875 g/L-hr , and wherein said fermentation production medium has a final pH that is lower than 3.86.2. A yeast strain of claim 1 , wherein said yeast strain produces lactic acid in a fermentation production medium with an average specific productivity of at least 3.00 g/L-hr.3. A yeast strain of claim 1 , wherein said yeast strain produces lactic acid in a fermentation production medium with an average specific productivity of at least 3.00 g/L-hr and a final pyruvic acid titer of less than 1 g/L.4Saccharomyces, Kluyveromyces, Issatchenkia, Pichia, Torulopsis, Candida, Hansenula, Zygosaccharomyces, Schizosaccharomyces, YarrowiaLachancea.. The yeast strain of claim 1 , wherein said yeast strain is selected from a group consisting of the genera and5SaccharomycesKluyveromyces.. The yeast strain of claim 1 , wherein said yeast strain is selected from a group consisting of the genera and6. The yeast strain of claim 1 , wherein said gene that encodes an exogenous lactate dehydrogenase is integrated at least at one chromosomal locus selected from a group consisting of a pyruvate decarboxylase gene or a homolog or analog thereof claim 1 , a phosphoenolpyruvate carboxykinse gene or a homolog or analog thereof claim 1 , a glycerol-3-phosphate phosphatase gene or a homolog or analog thereof claim 1 , and a NADH dehydrogenase 1 gene or a homolog or analog thereof.7. The yeast strain of claim 1 , wherein said gene that encodes an exogenous lactate dehydrogenase is ...

CHLOROGLOEOPSIS SP. HOST CELL FOR PRODUCING ETHANOL AND METHOD FOR PRODUCING ETHANOL USING THE SAME

One embodiment of the invention provides a genetically enhanced sp. host cell comprising at least one first recombinant gene encoding a first protein for the production of ethanol under the transcriptional control of a first inducible promoter, having at least 85%, 90% or 95% sequence identity to an endogenous inducible promoter of the sp. host cell. 2ChlorogloeopsisChlorogloeopsis fritschii, ChlorogloeopsisChlorogloeopsis. The genetically enhanced sp. host cell of claim 1 , wherein the host cell is PCC6912sp. PCC9212 claim 1 , or sp. ABICyano3.3ChlorogloeopsisChlorogloeopsis fritschii. The genetically enhanced sp. host cell of claim 2 , wherein the host cell is PCC6912.4Chlorogloeopsis. The genetically enhanced sp. host cell of claim 1 , further comprising at least one second recombinant gene encoding a second protein for the production of ethanol.5Chlorogloeopsis. The genetically enhanced sp. host cell of claim 1 , wherein the first recombinant gene encodes pyruvate decarboxylase.6Chlorogloeopsis. The genetically enhanced sp. host cell of claim 4 , wherein the second recombinant gene encodes alcohol dehydrogenase.7Chlorogloeopsis. The genetically enhanced sp. host cell of claim 1 , wherein the first recombinant gene encodes alcohol dehydrogenase E (AdhE) converting Acetyl-CoA into ethanol.8Chlorogloeopsis. The genetically enhanced sp. host cell of claim 4 , wherein both the first and second recombinant gene are under the transcriptional control of the same first endogenous inducible promoter.9Chlorogloeopsis. The genetically enhanced sp. host cell of claim 4 , wherein the first and second recombinant genes are under the transcriptional control of separate first and second promoters.10Chlorogloeopsis. The genetically enhanced sp. host cell of claim 9 , wherein the second promoter is a constitutive promoter.11Chlorogloeopsis. The genetically enhanced sp. host cell of claim 9 , wherein the second promoter is an inducible promoter.12. (canceled)13. (canceled) ...

RECOMBINANT ACID-RESISTANT YEAST IN WHICH ALCOHOL PRODUCTION IS INHIBITED AND METHOD FOR PRODUCING LACTIC ACID BY USING SAME

The present invention relates to: acid-resistant yeast to which lactic acid productivity is imparted, and in which the conversion of pyruvate into acetaldehyde is inhibited and, consequently, the ethanol production pathway is inhibited; and a method for producing lactic acid by using same. 1. A recombinant strain having lactic acid-producing ability , in which a pyruvate decarboxylase-encoding gene has been deleted or attenuated from an acid-tolerant yeast YBC strain (KCTC13508BP) and a lactate dehydrogenase-encoding gene is introduced into an acid-tolerant yeast YBC strain (KCTC13508BP).2. The recombinant strain of claim 1 , wherein the pyruvate decarboxylase-encoding gene is a g3002 gene.3. The recombinant strain of claim 2 , wherein the g3002 gene has the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2.4. The recombinant strain of claim 1 , wherein an alcohol dehydrogenase-encoding gene is additionally deleted.5. The recombinant strain of claim 4 , wherein the alcohol dehydrogenase-encoding gene is a g4423 gene.6. The recombinant strain of claim 1 , wherein the lactate dehydrogenase-encoding gene is introduced to replace the g3002 gene and is controlled by a promoter of the g3002 gene.7. The recombinant strain of claim 1 , which has reduced ethanol-producing ability compared to the YBC strain (KCTC13508BP) claim 1 , which is a parent strain claim 1 , due to deletion or attenuation of the g3002 gene.8. A method for producing lactic acid claim 1 , the method comprising steps of:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) producing lactic acid by culturing the recombinant strain of ; and'}(b) collecting the produced lactic acid.9. A gene which encodes a protein having pyruvate decarboxylase activity and having the amino acid sequence represented by SEQ ID NO: 3 or SEQ ID NO: 4.10. The gene of claim 9 , which has the nucleotide sequence represented by SEQ ID NO: 1 or SEQ ID NO: 2.11. A protein having pyruvate decarboxylase activity and having the amino ...

RECOMBINANT YEAST AND A METHOD FOR PRODUCING ETHANOL USING THE SAME

An acetic acid metabolizing ability of a recombinant yeast strain having xylose-metabolizing ability is to be improved. In such a recombinant yeast strain having xylose-metabolizing ability, the acetaldehyde dehydrogenase gene has been introduced and a gene encoding NADH dehydrogenase involved in reoxidation of cytoplasmic NADH on the mitochondrial outer membrane has been suppressed. 1. A recombinant yeast strain having xylose-metabolizing ability comprising the acetaldehyde dehydrogenase gene introduced thereinto , wherein a gene encoding NADH dehydrogenase involved in reoxidation of cytoplasmic NADH on the mitochondrial outer membrane is suppressed.2. The recombinant yeast strain according to claim 1 , wherein the gene encoding NADH dehydrogenase encodes a protein (a) or (b) below:(a) a protein comprising the amino acid sequence as shown in SEQ ID NO: 2 or 4; or{'sup': '+', '(b) a protein comprising an amino acid sequence exhibiting 70% or higher identity with the amino acid sequence as shown in SEQ ID NO: 2 or 4 and having enzymatic activity of catalyzing a reaction of converting NADH into NAD.'}3. The recombinant yeast strain according to comprising a xylose isomerase gene introduced thereinto.4. The recombinant yeast strain according to claim 3 , wherein the xylose isomerase gene encodes a protein (a) or (b) below:(a) a protein comprising the amino acid sequence as shown in SEQ ID NO: 6; or(b) a protein comprising an amino acid sequence exhibiting 70% or higher identity with the amino acid sequence as shown in SEQ ID NO: 6 and having enzymatic activity of converting xylose into xylulose.5. The recombinant yeast strain according to claim 1 , which further comprises a xylulokinase gene introduced thereinto.6. The recombinant yeast strain according to claim 1 , which comprises a gene encoding an enzyme selected from a group of enzymes constituting a non-oxidative process in the pentose phosphate pathway introduced thereinto.7. The recombinant yeast strain ...

HIGH YIELD ROUTE FOR THE PRODUCTION OF COMPOUNDS FROM RENEWABLE SOURCES

Provided herein are methods, compositions, and non-naturally occurring microbial organism for preparing compounds such as 1-butanol, butyric acid, succinic acid, 1,4-butanediol, 1-pentanol, pentanoic acid, glutaric acid, 1,5-pentanediol, 1-hexanol, hexanoic acid, adipic acid, 1,6-hexanediol, 6-hydroxy hexanoic acid, ε-Caprolactone, 6-amino-hexanoic acid, ε-Caprolactam, hexamethylenediamine, linear fatty acids and linear fatty alcohols that are between 7-25 carbons long, linear alkanes and linear α-alkenes that are between 6-24 carbons long, sebacic acid and dodecanedioic acid comprising: a) converting a Caldehyde and pyruvate to a Cβ-hydroxyketone intermediate through an aldol addition; and b) converting the Cβ-hydroxyketone intermediate to the compounds through enzymatic steps, or a combination of enzymatic and chemical steps. 1. A non-naturally occurring microbial organism comprising at least one exogenous nucleic acid encoding a 1 ,6-hexanediol pathway enzyme.2. The microbial organism of further comprising at least one enzyme selected from 2A wherein 2A is a 4-hydroxy-2-oxo-adipate aldolase claim 1 , or a 4 claim 1 ,6-dihydroxy-2-oxo-hexanoate aldolase.327-. (canceled)28. A non-naturally occurring microbial organism claim 1 , comprising at least one exogenous nucleic acid encoding a 1 claim 1 ,6-hexanediol pathway enzyme selected from 2A and one or more of 2B claim 1 , 3B1 claim 1 , 3B2 claim 1 , wherein 2A is a 4-hydroxy-2-oxo-adipate aldolase or a 4 claim 1 ,6-dihydroxy-2-oxo-hexanoate aldolase claim 1 , 2B is a 4-hydroxy-2-oxo-adipate dehydratase or a 4 claim 1 ,6-dihydroxy-2-oxo-hexanoate 4-dehydratase claim 1 , 3B1 is a 4-hydroxy-2-oxo-adipate 2-reductase or a 4 claim 1 ,6-dihydroxy-2-oxo-hexanoate 2-reductase claim 1 , and 3B2 is a 4-hydroxy-2-oxo-adipate 4-dehydrogenase or a 4 claim 1 ,6-dihydroxy-2-oxo-hexanoate 4-dehydrogenase.29. The organism of claim 28 , further comprising a 1 claim 28 ,6-hexanediol pathway enzyme selected from one or more of 2C claim ...

Polypeptides with Ketol-Acid Reductoisomerase Activity

Polypeptides having ketol-acid reductoisomerase activity are provided. Also disclosed are recombinant host cells comprising isobutanol biosynthetic pathways employing such polypeptides. Methods for producing isobutanol employing host cells comprising the polypeptides having ketol-acid reductoisomerase activity are also disclosed. 1. A recombinant host cell comprising an isobutanol biosynthetic pathway anda. a heterologous polypeptide with ketol-acid reductoisomerase activity having at least about 85%, at least about 90% identity, at least about 95%, or at least about 98% identity to one of the following: K9JM2 (SEQ ID NO: 193), K9JM3 (SEQ ID NO: 194), K9JM4 (SEQ ID NO: 195), K9JM5 (SEQ ID NO: 196), K9JM6 (SEQ ID NO: 197), K9JM7 (SEQ ID NO: 198), K9JM8 (SEQ ID NO: 199), K9JM9 (SEQ ID NO: 200), K9JM10 (SEQ ID NO: 201), K9JM11 (SEQ ID NO: 202), K9JM12 (SEQ ID NO: 203), K9JM13 (SEQ ID NO: 204), K9JM14 (SEQ ID NO: 205), K9JM15 (SEQ ID NO: 206), K9JM16 (SEQ ID NO: 207), K9JM17 (SEQ ID NO: 208), K9JM18 (SEQ ID NO: 209), K9JM19 (SEQ ID NO: 210), K9JM20 (SEQ ID NO: 211), K9JM21 (SEQ ID NO: 212), K9JM22 (SEQ ID NO: 213), K9JM23 (SEQ ID NO: 214), K9JM24 (SEQ ID NO: 215), K9JM25 (SEQ ID NO: 216), K9JM26 (SEQ ID NO: 217), K9JM27 (SEQ ID NO: 218), K9JM28 (SEQ ID NO: 219), K9JM29 (SEQ ID NO: 220), K9JM30 (SEQ ID NO: 221), K9JM31 (SEQ ID NO: 222), JM32 (SEQ ID NO: 223), JM33 (SEQ ID NO: 224), JM34 (SEQ ID NO: 225), JM35 (SEQ ID NO: 226), JM36 (SEQ ID NO: 227), JM37 (SEQ ID NO: 228), JM38 (SEQ ID NO: 229), JM39 (SEQ ID NO: 230), JM40 (SEQ ID NO: 231), JM42 (SEQ ID NO: 232), JM43 (SEQ ID NO: 233), JM44 (SEQ ID NO: 234), K9SB2 (SEQ ID NO: 235), K9_DAVID_SH (SEQ ID NO: 236), K9ALL3 (SEQ ID NO: 237), K9_URSALA (K9SB2+A56V) (SEQ ID NO: 239), JM41 (SEQ ID NO: 240), K9ALL148 (SEQ ID NO: 241), K9JM148 (SEQ ID NO: 242), K9ALL156 (SEQ ID NO: 243), K9JM156 (SEQ ID NO: 244), K9ALL191 (SEQ ID NO: 245), K9JM191 (SEQ ID NO: 246), K9ALL254 (SEQ ID NO: 247), K9ALL278 (SEQ ID NO: 248), K9ALL37 (SEQ ...

Microorganisms and methods for production of specific length fatty alcohols and related compounds

The invention provides non-naturally occurring microbial organisms containing a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length. Also provided are non-naturally occurring microbial organisms having a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms further include an acetyl-CoA pathway. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde or a fatty acid.

A BACTERIAL CELL FACTORY FOR EFFICIENT PRODUCTION OF ETHANOL FROM WHEY

The invention relates to a method for homo-ethanol production from lactose using a genetically modified lactic acid bacterium of the invention, where the cells are provided with a substrate comprising dairy waste supplemented with an amino nitrogen source (such as acid hydrolysed corn steep liquor). The invention further relates to genetically modified lactic acid bacterium and its use for homo-ethanol production from lactose in dairy waste. The lactic acid bacterium comprises both genes (lacABCD, LacEF, lacG) encoding enzymes catalysing the lactose catabolism pathway; and transgenes (pdc and adhB) encoding enzymes catalysing the conversion of pyruvate to ethanol. Additionally a number of genes (ldh, pta and adhE) are deleted in order to maximise homo-ethanol production as compared to production of lactate, acetoin and acetate production. 1. A method for ethanol production using a genetically engineered lactic acid bacterium comprising the steps of:a. introducing a genetically modified lactic acid bacterium into an aqueous culture medium;b. incubating the culture of (a);c. recovering ethanol produced by said culture during step (b), and optionally wherein the aqueous culture medium comprises:', 'I. whey permeate or residual whey permeate, and', wherein the genetically engineered lactic acid bacterium comprises transgenes encoding:', 'i. a polypeptide having pyruvate decarboxylase (PDC) activity (EC 4.1.1.1); and', 'wherein the genome of said lactic acid bacterium comprises genes encoding polypeptides having:', 'ii. a polypeptide having alcohol dehydrogenase B activity (EC 1.1.1.1); and'}, 'iii. lactose-specific phosphotransferase system (PTS) activity (EC 2.7.1.69)', 'iv. phospho-β-D-galactosidase activity (EC 3.2.1.85)', 'v. galactose-6-phosphate isomerase activity (EC 5.3.1.26),', 'vi. D-tagatose-6-phosphate kinase activity (EC 2.7.1.114), and', 'wherein the genome of said lactic acid bacterium is deleted for genes or lacks functional genes or genes encoding ...

Recombinant candida cell and preparation process and use thereof

Disclosed herein is a process for producing a recombinant Candida cell, which involves genetically engineering a parent Candida cell using a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated (Cas)(CRISPR/Cas) system. A recombinant Candida cell obtained using the process and a method for producing D-lactic acid from a biomass using the recombinant Candida cell are also disclosed.

Compositions and methods for 3-hydroxypropionate bio-production from biomass

Methods of obtaining mutant nucleic acid sequences that demonstrate elevated oxaloacetate a-decarboxylase activity are provided. Compositions, such as genetically modified microorganisms that comprise such mutant nucleic acid sequences, are described, as are methods to obtain the same.

Microorganisms and methods for producing pyruvate, ethanol, and other compounds

Microorganisms comprising modifications for producing pyruvate, ethanol, and other compounds. The microorganisms comprise modifications that reduce or ablate activity of one or more of pyruvate dehydrogenase, 2-oxoglutarate dehydrogenase, phosphate acetyltransferase, acetate kinase, pyruvate oxidase, lactate dehydrogenase, cytochrome terminal oxidase, succinate dehydrogenase, 6-phosphogluconate dehydrogenase, glutamate dehydrogenase, pyruvate formate lyase, pyruvate formate lyase activating enzyme, and isocitrate lyase. The microorganisms optionally comprise modifications that enhance expression or activity of pyruvate decarboxylase and alcohol dehydrogenase. The microorganisms are optionally evolved in defined media to enhance specific production of one or more compounds. Methods of producing compounds with the microorganisms are provided.

Use of Bacteriocin-Producing Ethanologens In Biofuel Production

An ethanologen for producing biofuel from one or more carbohydrates and reducing lactate and acetate production in a biofuel manufacturing process. The ethanologen is made by introducing into the ethanologen one or more exogenous genes required for production of a bacteriocin. The resulting ethanologen reduces lactate and acetate production by contaminant lactic acid bacteria by expression of the bacteriocin during the biofuel manufacturing process. Certain resulting ethanologens ferment sugars not naturally or not preferentially utilized by during the manufacturing process 1. An ethanologen for inhibiting contaminant lactic acid bacteria present in a biofuel manufacturing process , comprising an ethanologen containing one or more exogenous genes required for production of a bacteriocin , wherein production of the bacteriocin by the ethanologen inhibits contaminant lactic acid bacteria present in the biofuel manufacturing process.2. The ethanologen of claim 1 , wherein inhibition of the contaminant lactic acid bacteria results in reduced lactate and acetate levels in the biofuel manufacturing process.3. The ethanologen of claim 1 , wherein the ethanologen is capable of fermenting sugars not naturally or not preferentially utilized by a main fermenting microbe present in the biofuel manufacturing process.4Saccharomyces cerevisiae.. The ethanologen of claim 3 , wherein the main fermenting microbe is5. The ethanologen of claim 1 , wherein the ethanologen is a native biofuel-producing organism.6SacchoromycesZymononas. The ethanologen of claim 5 , wherein the native biofuel-producing organism is sp. or sp.7. The ethanologen of claim 1 , wherein the ethanologen is an organism engineered to produce the biofuel.8EscherichiaClostridium. The ethanologen of claim 7 , wherein the organism engineered to produce the biofuel is a lactic acid bacterium claim 7 , sp. claim 7 , or sp.9LactobacillusLactococcusEnterococcusStreptococcus. The ethanologen of claim 8 , wherein the lactic ...

MICROORGANISMS AND METHODS FOR PRODUCING PYRUVATE, ETHANOL, AND OTHER COMPOUNDS

Microorganisms comprising modifications for producing pyruvate, ethanol, and other compounds. The microorganisms comprise modifications that reduce or ablate activity of one or more of pyruvate dehydrogenase, 2-oxoglutarate dehydrogenase, phosphate acetyltransferase, acetate kinase, pyruvate oxidase, lactate dehydrogenase, cytochrome terminal oxidase, succinate dehydrogenase, 6-phosphogluconate dehydrogenase, glutamate dehydrogenase, pyruvate formate lyase, pyruvate formate lyase activating enzyme, and isocitrate lyase. The microorganisms optionally comprise modifications that enhance expression or activity of pyruvate decarboxylase and alcohol dehydrogenase. The microorganisms are optionally evolved in defined media to enhance specific production of one or more compounds. Methods of producing compounds with the microorganisms are provided. 120-. (canceled)21. A microorganism comprising activity-reducing or activity-ablating mutations in endogenous genes encoding:one or more enzymes in a first set selected from the group consisting of pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase;one or more enzymes in a second set selected from the group consisting of phosphate acetyltransferase, acetate kinase, and pyruvate oxidase; and lactate dehydrogenase and one or more enzymes selected from the group consisting of cytochrome terminal oxidase and succinate dehydrogenase; or', 'one or more enzymes selected from the group consisting of cytochrome terminal oxidase and succinate dehydrogenase and one or more enzymes selected from the group consisting of 6-phosphogluconate dehydrogenase and glutamate dehydrogenase., 'enzymes in a third set comprising22. The microorganism of claim 21 , wherein the one or more enzymes in the first set is pyruvate dehydrogenase.23. The microorganism of claim 21 , wherein the one or more enzymes in the second set are selected from the group consisting of phosphate acetyltransferase and pyruvate oxidase.24. The microorganism of claim 21 , ...

Recombinant Host Cells Comprising Phosphoketalase

The present invention is related to recombinant host cells comprising: (i) at least one deletion, mutation, and/or substitution in an endogenous gene encoding a polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA; and (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. The present invention is also related to recombinant host cells further comprising (iii) a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.

Protein manipulation

A method of improving the folding of an enzyme comprising a thiamine pyrophosphate (TPP) binding domain, the method comprising: providing a nucleic acid encoding the enzyme comprising a TPP binding domain, in which one or more of the TPP binding domains in the enzyme monomer are replaced with a TPP binding domain from a thermostable TPP-binding protein, and expressing the nucleic acid under conditions that allow expression and folding of the enzyme. The enzyme may be pyruvate decarboxylase.

Bacteria engineered to treat disorders involving the catabolism of a branched chain amino acid

The present disclosure provides recombinant bacterial cells that have been engineered with genetic circuitry which allow the recombinant bacterial cells to sense a patient's internal environment and respond by turning an engineered metabolic pathway on or off. When turned on, the recombinant bacterial cells complete all of the steps in a metabolic pathway to achieve a therapeutic effect in a host subject. These recombinant bacterial cells are designed to drive therapeutic effects throughout the body of a host from a point of origin of the microbiome. Specifically, the present disclosure provides recombinant bacterial cells comprising a heterologous gene encoding a branched chain amino acid catabolism enzyme. The disclosure further provides pharmaceutical compositions comprising the recombinant bacteria, and methods for treating disorders involving the catabolism of branched chain amino acids using the pharmaceutical compositions disclosed herein.

Host Cells and Methods for Production of Isobutanol

The invention relates to recombinant host cells having at least one integrated polynucleotide encoding a polypeptide that catalyzes a step in a pyruvate-utilizing biosynthetic pathway, e.g., pyruvate to acetolactate conversion. The invention also relates to methods of increasing the biosynthetic production of isobutanol, 2,3-butanediol, 2-butanol or 2-butanone using such host cells. 1. A recombinant host cell comprising:{'i': Bacillus subtilis, Klebsiella pneumonia, Lactococcus lactis, Staphylococcus aureus, Listeria monocytogenes, Streptococcus mutans, Streptococcus thermophiles, Vibrio angustum', 'Bacillus cereus;, '(a) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of pyruvate to acetolactate wherein the polypeptide is an acetolactate synthase from , or'}(b) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of acetolactate to 2,3-dihydroxyisovalerate wherein the polypeptide is a ketol-acid reductoisomerase and the ketol-acid reductoisomerase has at least 95% identity to SEQ ID NO: 224;{'i': Escherichia coli, Bacillus subtilis, Methanococcus maripaludis', 'Streptococcus mutans;, '(c) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of 2,3-dihydroxyisovalerate to α-ketoisovalerate wherein the polypeptide is a dihydroxyacid dehydratase from , or'}(d) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of α-ketoisovalerate to isobutyraldehyde wherein the polypeptide is a branched-chain α-keto acid decarboxylase and the branched-chain α-keto acid decarboxylase has at least 95% identity to SEQ ID NO: 48; and{'i': Achromobacter xylosoxidans', 'Beijerinkia indica,, '(e) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion isobutyraldehyde to isobutanol wherein the polypeptide is an alcohol dehydrogenase from or'}wherein expression of pyruvate decarboxylase in the ...

TROPANE ALKALOID (TA) PRODUCING NON-PLANT HOST CELLS, AND METHODS OF MAKING AND USING THE SAME

Provided herein, among other things, is an engineered non-plant cell that produces a tropane alkaloid product, a precursor of a tropane alkaloid product, or a derivative of a tropane alkaloid product. A method for producing a tropane alkaloid, a precursor of a tropane alkaloid product, or a derivative of a tropane alkaloid product that makes use of the cell is also described. 1. An engineered non-plant cell that produces a precursor of a tropane alkaloid product , a tropane alkaloid product , or a derivative of a tropane alkaloid product , wherein the engineered non-plant cell comprises a plurality of heterologous coding sequences encoding a plurality of enzymes within a pathway for producing the precursor of a tropane alkaloid product , the tropane alkaloid product , or the derivative of a tropane alkaloid product;wherein the cell comprises one or more alterations to one or more endogenous metabolic pathways or regulatory mechanisms selected from the group of endogenous arginine metabolism, endogenous phenylalanine and phenylpropanoid metabolism, endogenous polyamine regulatory mechanisms and metabolism, endogenous acetate metabolism, and endogenous glycoside metabolism.2. The cell of claim 1 , wherein the cell comprises one or more alterations to one or more endogenous metabolic pathways or regulatory mechanisms selected from the group of endogenous arginine metabolism claim 1 , endogenous phenylalanine and phenylpropanoid metabolism claim 1 , endogenous polyamine regulatory mechanisms and metabolism claim 1 , and endogenous acetate metabolism.3. The cell of claim 1 , wherein the cell comprises one or more alterations to endogenous glycoside metabolism4. The cell of - claim 1 , wherein the cell is a microbial cell.5. The cell of claim 4 , wherein the cell is a fungal cell.6. The cell of - claim 4 , wherein the engineered cell comprises one or more heterologous coding sequences for one or more enzymes claim 4 , wherein at least one of the enzymes is selected from ...

CYANOBACTERIAL STRAINS CAPABLE OF UTILIZING PHOSPHITE

The invention provides genetically modified cyanobacterial cells that are capable of utilizing phosphite as a primary phosphorus source, and can out-compete contaminant organisms for certain forms of phosphorus more effectively. 1. A genetically modified cyanobacterial cell for the production of a product of interest , comprising:a) at least one recombinant gene that encodes a heterologous phosphite dehydrogenase enzyme EC:1.20.1.1 that catalyzes the oxidation of phosphite to phosphate;b) an operon comprising at least one recombinant phosphite transporter gene encoding at least one phosphite transporter protein for transporting phosphite into the cell; andc) at least one recombinant production gene encoding a polypeptide for the production of said product of interest.2CyanotheceRalstonia. The genetically modified cyanobacterial cell of claim 1 , wherein the recombinant phosphite dehydrogenase gene encodes a polypeptide that has a sequence identity of greater than 60% to the protein sequence of the phosphite dehydrogenase enzyme from (SEQ ID NO: 16) or from (SEQ ID NO: 12).3. The genetically modified cyanobacterial cell of claim 1 , wherein the recombinant phosphite dehydrogenase gene is operably linked to a regulatable promoter selected from the group consisting of: a metal-regulatable promoter claim 1 , a nitrate-regulatable promoter claim 1 , and a phosphorus-regulatable promoter.4. The genetically modified cyanobacterial cell of claim 1 , wherein at least one of the phosphite transporter genes is derived from a different organism than the phosphite dehydrogenase gene.5Desulfotignum phosphitoxidansCyanothece.. The genetically modified cyanobacterial cell of claim 1 , wherein the at least one phosphite transporter gene is from or6. The genetically modified cyanobacterial cell of claim 1 , wherein the at least one phosphite transporter protein is selected from the group consisting of PtxA claim 1 , PtxB claim 1 , PtxC claim 1 , and PtdC.7. The genetically modified ...

System and method of optogenetically controlling metabolic pathways for the production of chemicals

A system and method for controlling metabolic enzymes or pathways in cells to produce a chemical above the levels of a wild-type strain is disclosed. The system utilizes cells, including yeasts, bacteria, and molds, having at least two genes capable of being controlled bi-directionally with light, where one gene is turned from off to on when exposed to light and another gene is turned from on to off when exposed to light, the two genes reversing when the light is turned off. Cells may utilize any number of sequences that benefit chemical production, including sequences that: encode for constitutive transcription of light-activated transcription factor fusions; encode for a metabolic enzyme; encode for a repressor; induce expression of metabolic enzymes; and an endogenous or exogenous activator expressed by a constitutive promoter, inducible promoter, or gene circuit. These systems may be coupled to biosensors or protein cascade systems, enabling the monitoring or automation of the fermentation process to optimize production of a desired product. These systems may also allow for optimization and periodic operation of a bioreactor using light pulses.

Integration of a Polynucleotide Encoding a Polypeptide that Catalyzes Pyruvate to Acetolactate Conversion

The invention relates to recombinant host cells having at least one integrated polynucleotide encoding a polypeptide that catalyzes a step in a pyruvate-utilizing biosynthetic pathway, e.g., pyruvate to acetolactate conversion. The invention also relates to methods of increasing the biosynthetic production of isobutanol, 2,3-butanediol, 2-butanol or 2-butanone using such host cells. 147-. (canceled)48. A recombinant host cell comprising:{'i': Bacillus subtilis, Klebsiella pneumonia, Lactococcus lactis, Staphylococcus aureus, Listeria monocytogenes, Streptococcus mutans, Streptococcus thermophiles, Vibrio angustum', 'Bacillus cereus., '(a) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of pyruvate to acetolactate wherein the polypeptide is an acetolactate synthase from , or'}{'i': Lactococcus lactis, Vibrio cholera, Pseudomonas aeruginosa, Pseudomonas fluorescens', 'Anaerostipes caccae., '(b) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of acetolactate to 2,3-dihydroxyisovalerate wherein the polypeptide is a ketol-acid reductoisomerase from , or'}{'i': Escherichia coli, Bacillus subtilis, Methanococcus maripaludis', 'Streptococcus mutans, '(c) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of 2,3-dihydroxyisovalerate to α-ketoisovalerate wherein the polypeptide is a dihydroxyacid dehydratase from , or ; and'}{'i': Listeria grayi, Lactococcus lactis', 'Macrococcus caseolyticus., '(d) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of α-ketoisovalerate to isobutyraldehyde wherein the polypeptide is a branched-chain α-keto acid decarboxylase from , or'}49Achromobacter xylosoxidansBeijerinkia indica.. The recombinant host cell of further comprising a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion isobutyraldehyde to isobutanol wherein the polypeptide ...

METHODS FOR THE POSITIVE SELECTION OF ETHANOL OVERPRODUCING MUTANTS FROM SACCHAROMYCES CEREVISIAE

Described herein are new approaches for the selection of strains with increased ethanol production from hydrolyzed starch derived sugars. An industrial production strain of AS400 was subjected to positive selection of mutants resistant to toxic concentrations of oxythiamine, trehalose, 3-bromopyruvate, glyoxylic acid, and glucosamine. The selected mutants are characterized by 5-8% increase in ethanol yield (g gof consumed glucose) as compared to the parental industrial ethanol-producing strain. A multiple-step selection approach that consisted of the sequential selection using glyoxylic acid, glucosamine and bromopyruvate as selective agents resulted in a 12% increase in ethanol yield during fermentation on industrial media. These results indicate that the selection methods provided herein are useful for producing a variety of strains that are promising candidates for industrial ethanol production. 1S. cerevisiae. A method of making a strain with enhanced ethanol producing characteristics comprising ,{'i': 'S. cerevisiae', 'contacting a culture of a parent strain of with a sufficient amount of a first selection agent selected from the group consisting of oxythiamine, trehalose, bromopyruvate, glycoxylic acid and glucosamine for the selection agent to be toxic to the parent strain;'}growing the contacted culture on a plating medium containing a sugar as a carbon source;selecting candidate spontaneous mutant strains of the culture that grow on the plating medium, and{'i': 'S. cerevisiae', 'measuring an ethanol producing characteristic of the candidate spontaneous mutant strains to determine whether the candidate strain demonstrates enhanced ethanol production characteristics in comparison to the parent strain and if so, obtaining the strain with enhanced ethanol producing characteristics.'}2. The method of wherein the obtained spontaneous mutant strain with enhanced ethanol production characteristics is contacted with an amount of a second selection agent selected ...

METHOD FOR PRODUCING LACTIC ACID

The present invention provides a method for producing lactic acid in a recombinant yeast cell culture using glucose as carbon source comprising a first, seed fermentation stage to produce biomass wherein the yeast is cultivated in a culture medium at a pH of 5 to 7, followed by a second, a production fermentation stage with biomass from the seed fermentation to produce lactic acid, wherein the yeast is cultivated in a culture medium at low p H using a yeast strain that is engineered to have lactate dehydrogenase (LDH) activity and optionally has decreased or knocked-out pyruvate decarboxylase (PDC) activity. 1. A method for producing lactic acid in a recombinant yeast cell culture using glucose as carbon source comprising:a) cultivating yeast in a seed fermentation stage to produce biomass wherein the yeast is cultivated in a culture medium at a pH of 5 to 7, followed byb) cultivating yeast in a production fermentation stage with biomass from the seed fermentation stage to produce lactic acid, wherein the yeast is cultivated in a culture medium at a pH of <5, andwherein said yeast has lactate dehydrogenase (LDH) activity and optionally decreased pyruvate decarboxylase (PDC) activity.2. The method according to claim 1 , wherein the yeast is a diploid yeast.3. The method according to claim 1 , wherein the yeast is a polyploid or aneuploid yeast.4. The method according to claim 1 , wherein the production fermentation stage is at a pH of <4.5 claim 1 , <4 claim 1 , or <3.5 claim 1 , and wherein the production fermentation stage has a final pH of 3 or less claim 1 , <2.9 claim 1 , <2.8 claim 1 , <2.7 claim 1 , <2.6 claim 1 , <2.5 claim 1 , <2.4 claim 1 , <2.3 claim 1 , <2.2 claim 1 , or <2.15.5. The method according to claim 1 , wherein the seed fermentation stage is performed under fed-batch conditions and/or the production fermentation stage is performed under batch process conditions claim 1 , and wherein the seed fermentation stage and the production fermentation ...

Ethanol Production in Microorganisms

The present disclosure relates to methods and compositions for engineering photoautotrophic organisms to convert carbon dioxide and light into fatty acid esters and other molecules, including biofuels. The molecules are then secreted by the organism into a growth medium. 1. A method for production of ethanol , comprising:culturing an engineered photosynthetic microbe in a culture medium in the presence of light and inorganic carbon, wherein the engineered photosynthetic microbe comprises a recombinant pyruvate decarboxylase nucleic acid sequence encoding a pyruvate decarboxylase having EC 4.1.1.1 and a recombinant alcohol dehydrogenase nucleic acid sequence encoding an alcohol dehydrogenase having EC 1.1.1.1 or EC 1.1.1.2, wherein the copy number of at least the alcohol dehydrogenase nucleic acid sequence in the engineered photosynthetic microbe is greater than the copy number of an alcohol dehydrogenase nucleic acid sequence in a control engineered photosynthetic microbe comprising a single recombinant alcohol dehydrogenase gene and a single recombinant pyruvate decarboxylase gene regulated together by a single promoter, and wherein the engineered photosynthetic microbe produces ethanol in an amount greater than a non-engineered photosynthetic microbe, when cultured under identical conditions.2. The method of claim 1 , wherein different promoters are used to achieve differential expression of each enzyme.3. The method of claim 2 , wherein one of the recombinant nucleic acid sequences is modulated by an inducible promoter and one of the recombinant nucleic acid sequences is modulated by a constitutive promoter.4. The method of claim 3 , wherein the copy number of the recombinant alcohol dehydrogenase nucleic acid sequence is modulated by a constitutive promoter.5. The method of claim 3 , wherein the copy number of the recombinant pyruvate decarboxylase nucleic acid sequence is modulated by a chemically-affected promoter.6. The method of claim 5 , wherein the copy ...

ELECTRON CONSUMING ETHANOL PRODUCTION PATHWAY TO DISPLACE GLYCEROL FORMATION IN S. CEREVISIAE

The present invention provides for a mechanism to completely replace the electron accepting function of glycerol formation with an alternative pathway to ethanol formation, thereby reducing glycerol production and increasing ethanol production. In some embodiments, the invention provides for a recombinant microorganism comprising a down-regulation in one or more native enzymes in the glycerol-production pathway. In some embodiments, the invention provides for a recombinant microorganism comprising an up-regulation in one or more enzymes in the ethanol-production pathway. 160-. (canceled)61. A co-culture comprising at least two host cells wherein (i) a heterologous nucleic acid encoding a phosphoketolase;', '(ii) at least one heterologous nucleic acid encoding an enzyme in an acetyl-CoA production pathway;', '(iii) a heterologous nucleic acid encoding a bifunctional acetaldehyde-alcohol dehydrogenase; and,', '(iv) at least one genetic modification that leads to the down-regulation of an enzyme in a glycerol-production pathway; and,, '(a) one of the host cells comprises(b) another host cell that is genetically distinct from (a).62. The co-culture of claim 61 , wherein the host cell is a yeast and the genetically distinct host cell is a yeast or bacterium.63. The recombinant microorganism of claim 61 , wherein said phosphoketolase is a single-specificity phosphoketolase with the Enzyme Commission Number 4.1.2.9.64. The recombinant microorganism of claim 61 , wherein said phosphoketolase is dual-specificity phosphoketolase with the Enzyme Commission Number 4.1.2.22.65Aspergillus, Neurospora, Lactobacillus, Bifidobacterium, Penicillium, LeuconostocOenococcus.. The recombinant microorganism of claim 61 , wherein said phosphoketolase is from a genus selected from the group consisting of claim 61 , and66. The recombinant microorganism of claim 61 , wherein said phosphoketolase corresponds to a polypeptide selected from a group consisting of SEQ ID NOs: 9 claim 61 , 11 claim ...

Ethanol Production in Microorganisms

The present disclosure relates to methods and compositions for engineering photoautotrophic organisms to convert carbon dioxide and light into fatty acid esters and other molecules, including biofuels. The molecules are then secreted by the organism into a growth medium. 1. A method for production of ethanol , comprising:culturing an engineered photosynthetic microbe in a culture medium in the presence of light and inorganic carbon, wherein said engineered photosynthetic microbe comprising a recombinant pyruvate decarboxylase nucleic acid sequence encoding a pyruvate decarboxylase and a recombinant alcohol dehydrogenase nucleic acid sequence encoding an alcohol dehydrogenase, wherein the expression level the alcohol dehydrogenase and the expression level of the pyruvate decarboxylase are regulated by separate promoters, wherein the expression level of alcohol dehydrogenase in the engineered photosynthetic microbe is greater than the expression level of alcohol dehydrogenase in a control engineered photosynthetic microbe comprising a recombinant alcohol dehydrogenase nucleic acid sequence and a recombinant pyruvate decarboxylase nucleic acid sequence regulated together by a single promoter, and wherein the engineered photosynthetic microbe produces ethanol in an amount greater than a non-engineered photosynthetic microbe, when cultured under identical conditions.2. The method of claim 1 , wherein different promoters are used to achieve differential expression of each recombinant nucleic acid sequence.3. The method of claim 2 , wherein one of the recombinant nucleic acid sequences is modulated by an inducible promoter and one of the recombinant nucleic acid sequences is modulated by a constitutive promoter.4. The method of claim 3 , wherein the expression of the recombinant alcohol dehydrogenase nucleic acid sequence is modulated by a constitutive promoter.5. The method of claim 3 , wherein the expression of the recombinant pyruvate decarboxylase nucleic acid sequence ...

TRANSFORMANT AND PROCESS FOR PRODUCTION THEREOF, AND PROCESS FOR PRODUCTION OF LACTIC ACID

The present invention relates to a transformant which uses as a host into which a D-LDH gene derived from bacteria of the genus and a D-LDH gene derived from bacteria of the genus are incorporated and in which some of the genes in a group of pyruvate decarboxylase-encoding genes of the host have been deleted or inactivated. 1Schizosaccharomyces pombePediococcusLactobacillus. A transformant which uses as a host into which a D-lactate dehydrogenase gene derived from bacteria of the genus and a D-lactate dehydrogenase gene derived from bacteria of the genus are incorporated ,{'i': 'Schizosaccharomyces pombe', 'wherein some of the genes in a group of pyruvate decarboxylase-encoding genes of the host have been deleted or inactivated.'}2. The transformant according to claim 1 ,{'i': Pediococcus', 'Pediococcus acidilactici', 'Pediococcus pentosaceus, 'wherein the bacteria of the genus are or , and'}{'i': Lactobacillus', 'Lactobacillus pentosus, Lactobacillus bulgaricus', 'Lactobacillus brevis., 'the bacteria of the genus are , or'}3. The transformant according to or claim 1 ,wherein the deleted or inactivated genes in the group of pyruvate decarboxylase-encoding genes are PDC2 genes.4. The transformant according to any one of to claim 1 ,{'i': 'Schizosaccharomyces pombe.', 'wherein the D-lactate dehydrogenase gene is incorporated into a chromosome of the'}5Schizosaccharomyces pombePediococcusLactobacillusSchizosaccharomyces pombe. A process for production of a transformant using as a host into which a D-lactate dehydrogenase gene derived from bacteria of the genus and a D-lactate dehydrogenase gene derived from bacteria of the genus are incorporated and in which some of the genes in a group of pyruvate decarboxylase-encoding genes of the host have been deleted or inactivated claim 1 , the process comprising:a step of obtaining a transformant by introducing an expression cassette into the host,{'i': Schizosaccharomyces pombe', 'Pediococcus', 'Schizosaccharomyces pombe', ' ...

Lactic acid bacteria for the production of ethanol from biomass material

Lactic acid bacterial cultures, cell populations and articles of manufacture comprising same are disclosed for generating ethanol from lignocellulse.

METHOD FOR PRODUCING ACETOIN

The present invention relates to a recombinant yeast having a reduced pyruvate decarboxylase activity, in the genome of which has been inserted:—one or more nucleic acids encoding an acetolactate synthase or ALS,—one or more nucleic acids en coding an acetolactate decarboxylase or ALD, and—one or more copies of a nucleic acids encoding a NADH oxidase or NOXE. 1. A recombinant yeast having a reduced pyruvate decarboxylase activity , in the genome of which has been inserted:one or more nucleic acids encoding an acetolactate synthase or ALS,one or more nucleic acids encoding an acetolactate decarboxylase or ALD, andone or more copies of a nucleic acids encoding a NADH oxidase or NOXE.2. The recombinant yeast according to claim 1 , wherein the said recombinant yeast comprises one or more DNA constructs selected from a group comprising the following formulae:{'br': None, 'sub': x1', 'x2', 'x3, '5′-[Gene 1]-3′ and 5′-[Gene 2]-3′ and 5′-[Gene 3]-3′, \u2003\u2003(I)'}{'br': None, 'sub': x1', 'x2', 'x3, '5′-[Gene 1]-[Gene 2]-3′ and 5′-[Gene 3]-3′, \u2003\u2003(II)'}{'br': None, 'sub': x1', 'x2', 'x3, '5′-[Gene 1]-[Gene 2]-[Gene 3]-3′, and \u2003\u2003(III)'}a combination thereof,wherein:“Gene 1” means a nucleic acid selected from a group comprising ALS, ALD or NOXE;“Gene 2” means a nucleic acid selected from a group comprising ALS, ALD or NOXE but different from gene 1;“Gene 3” means a nucleic acid selected from a group comprising ALS, ALD or NOXE but different from genes 1 and 2;“ALS” is a nucleic acid encoding an acetolactate synthase;“ALD” is a nucleic acid encoding an acetolactate decarboxylase;“NOXE” is a nucleic acid encoding a NADH oxidase;each of “x1”, “x2” and “x3”, one independently from the others, represents an integer ranging from 0 to 50, andprovided that said recombinant yeast comprises at least one nucleic acid encoding for each of ALS, ALD and NOXE.3. The recombinant yeast according to claim 2 , wherein the said recombinant yeast comprises at least one DNA ...

Yeast cell having reduced ethanol productivity and use of the yeast cell

A yeast cell, in which an alcohol dehydrogenase 6 activity is increased and an activity of converting acetaldehyde to ethanol is decreased, a method of decreasing ethanol production by using the yeast cell, and a method of producing lactate.

3-HYDROXYPROPIONIC ACID PRODUCTION BY RECOMBINANT YEASTS EXPRESSING AN INSECT ASPARTATE 1-DECARBOXYLASE

Provided herein are recombinant yeast cells having an active 3-Hydroxypropionic Acid (3-HP) pathway and further comprising a heterologous polynucleotide encoding an aspartate 1-decarboxylase (ADC) of the Class Insecta, Bivalvia, Branchioporia, Gastropoda, or Leptocardii. Also described are methods of using the recombinant yeast cells to produce 3-HP and acrylic acid. 1. A recombinant yeast cell comprising a heterologous polynucleotide encoding an aspartate 1-decarboxylase of the Class Insecta , Bivalvia , Branchiopoda , Gastropoda , or Leptocardii , wherein the cell is capable of producing 3-hydroxypropionic acid (3-HP).2. The recombinant yeast cell of claim 1 , wherein the aspartate 1-decarboxylase of the Class Insecta is an insect aspartate 1-decarboxylase of an Order selected from the group consisting of Blattodea claim 1 , Coleoptera claim 1 , Dermaptera claim 1 , Diptera claim 1 , Embiidina claim 1 , Ephemeroptera claim 1 , Hemiptera claim 1 , Hymenoptera claim 1 , Lepidoptera claim 1 , Mantoptera claim 1 , Mecoptera claim 1 , Megaloptera claim 1 , Microcoryphia claim 1 , Neuroptera claim 1 , Notoptera claim 1 , Odonata claim 1 , Orthoptera claim 1 , Phasmatodea claim 1 , Plecoptera claim 1 , Psocoptera claim 1 , Raphidioptera claim 1 , Siphonaptera claim 1 , Strepsiptera claim 1 , Thysanoptera claim 1 , Trichoptera claim 1 , Zoraptera claim 1 , and Zygentoma.3. The recombinant yeast cell of claim 1 , wherein the aspartate 1-decarboxylase of the Class Insecta has at least 60% claim 1 , sequence identity to the amino acid sequence of SEQ ID NO: 162 claim 1 , 163 claim 1 , 164 claim 1 , 165 claim 1 , 166 claim 1 , 167 claim 1 , 168 claim 1 , 169 claim 1 , 170 claim 1 , 171 claim 1 , 172 claim 1 , 173 claim 1 , 174 claim 1 , 175 claim 1 , 176 claim 1 , 177 claim 1 , 178 claim 1 , 179 claim 1 , 180 claim 1 , 181 claim 1 , 182 claim 1 , 183 claim 1 , 184 claim 1 , 185 claim 1 , 186 claim 1 , 187 claim 1 , 188 claim 1 , 189 claim 1 , 190 claim 1 , or 191.4. The ...

MICROORGANISM STRAINS FOR THE PRODUCTION OF 2.3- BUTANEDIOL

A recombinant yeast having a reduced pyruvate decarboxylase activity, in the génome of which has been inserted: —one or more nucleic acids encoding an acetolactate synthase or ALS, —one or more nucleic acids encoding an acetolactate decarboxylase or ALD, —one or more nucleic acids encoding a butancdiol dehydrogenase or BDH, and —one or more copies of a nucleic acids encoding a NADH oxidase or NOXE. 1. A recombinant yeast having a reduced pyruvate decarboxylase activity , in the genome of which has been inserted:one or more nucleic acids encoding an acetolactate synthase or ALS,one or more nucleic acids encoding an acetolactate decarboxylase or ALD,one or more nucleic acids encoding a butanediol dehydrogenase or BDH, andone or more copies of a nucleic acids encoding a NADH oxidase or NOXE.2. The recombinant yeast according to claim 1 , wherein the said recombinant yeast comprises one or more DNA constructs selected in a group comprising the following formulae:{'br': None, 'sub': x1', 'x2', 'x3', 'x4, '5′-[Gene 1]-3′ and 5′-[Gene 2]-3′ and 5′-[Gene 3]-3′ and 5′-[Gene 4]-3′,\u2003\u2003(I)'}{'br': None, 'sub': x1', 'x2', 'x3', 'x4, '5′-[Gene 1]-[Gene 2]-[Gene 3]-3′ and 5′-[Gene 4]-3′,\u2003\u2003(II)'}{'br': None, 'sub': x1', 'x2', 'x3', 'x4, '5′-[Gene 1]-[Gene 2]-3′ and 5′-[Gene 3]-[Gene 4]-3′,\u2003\u2003(III)'}{'br': None, 'sub': x1', 'x2', 'x3', 'x4, '5′-[Gene 1]-[Gene 2]-[Gene 3]-[Gene 4]-3′, and\u2003\u2003(IV)'}a combination thereof, “Gene 1” means a nucleic acid selected from a group comprising ALS, ALD, BDH or NOXE;', '“Gene 2” means a nucleic acid selected from a group comprising ALS, ALD, BDH or NOXE but different from gene 1;', '“Gene 3” means a nucleic acid selected from a group comprising ALS, ALD, BDH or NOXE but different from genes 1 and 2;', '“Gene 4” means a nucleic acid selected from a group comprising ALS, ALD, BDH or NOXE but different from genes 1 to 3;', '“ALS” is a nucleic acid encoding an acetolactate synthase;', '“ALD” is a nucleic acid ...

ELECTRON CONSUMING ETHANOL PRODUCTION PATHWAY TO DISPLACE GLYCEROL FORMATION IN S. CEREVISIAE

The present invention provides for a mechanism to completely replace the electron accepting function of glycerol formation with an alternative pathway to ethanol formation, thereby reducing glycerol production and increasing ethanol production. In some embodiments, the invention provides for a recombinant microorganism comprising a down-regulation in one or more native enzymes in the glycerol-production pathway. In some embodiments, the invention provides for a recombinant microorganism comprising an up-regulation in one or more enzymes in the ethanol-production pathway. 1. A co-culture comprising at least two host cells wherein (i) a heterologous nucleic acid encoding a phosphoketolase;', '(ii) at least one heterologous nucleic acid encoding an enzyme in an acetyl-CoA production pathway;', '(iii) a heterologous nucleic acid encoding a bifunctional acetaldehyde-alcohol dehydrogenase; and,', '(iv) at least one genetic modification that leads to the down-regulation of an enzyme in a glycerol-production pathway; and,, '(a) one of the host cells comprises(b) another host cell that is genetically distinct from (a).2. The co-culture of claim 1 , wherein the host cell is a yeast and the genetically distinct host cell is a yeast or bacterium.3. The recombinant microorganism of claim 1 , wherein said phosphoketolase is a single-specificity phosphoketolase with the Enzyme Commission Number 4.1.2.9.4. The recombinant microorganism of claim 1 , wherein said phosphoketolase is dual-specificity phosphoketolase with the Enzyme Commission Number 4.1.2.22.5Aspergillus, Neurospora, Lactobacillus, Bificlobacterium, Penicillium, Leuconostoc,Oenococcus.. The recombinant microorganism of claim 1 , wherein said phosphoketolase is from a genus selected from the group consisting of and6. The recombinant microorganism of claim 1 , wherein said phosphoketolase corresponds to a polypeptide selected from a group consisting of SEQ ID NOs: 9 claim 1 , 11 claim 1 , 12 claim 1 , 13 claim 1 , 14 ...

MICROORGANISMS AND METHODS FOR PRODUCTION OF SPECIFIC LENGTH FATTY ALCOHOLS AND RELATED COMPOUNDS

The invention provides non-naturally occurring microbial organisms containing a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length. Also provided are non-naturally occurring microbial organisms having a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms further include an acetyl-CoA pathway. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde or a fatty acid. 126.-. (canceled)28. The non-naturally occurring microbial organism of claim 27 , wherein said microbial organism comprises:(a) two, three, or four exogenous nucleic acids each encoding an enzyme of said MI-FAE cycle;(b) two, three, or four exogenous nucleic acids each encoding an enzyme of said termination pathway; or(c) exogenous nucleic acids encoding each of the enzymes of at least one of the pathways selected from (1)-(13).29. The non-naturally occurring microbial organism of claim 27 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.30. The non-naturally occurring microbial organism of claim 27 , wherein said microbial organism further comprises an acetyl-CoA pathway and at least one exogenous nucleic acid encoding an acetyl-CoA pathway enzyme expressed in a sufficient amount to produce acetyl-CoA claim 27 , wherein said acetyl-CoA pathway comprises a pathway selected from:(1) 2A and 2B;(2) 2A, 2C, and 2D;(3) 2H;(4) 2G and 2D;(5) 2E, 2F and 2B;(6) 2E and 21;(7) 2J, 2F and 2B;(8) 2J and 2I;(9) 3A, 3B, and 3C;(10) 3A, 3B, 3J, 3K, and 3D;(11) 3A, 3B, 3G, and 3D;(12) 3A, 3F, and 3D;(13) 3N, 3H, 3B and 3C;(14) 3N, 3H, 3B, 3J, 3K, and 3D;(15) 3N, 3H, 3B, 3G, and 3D;(16) ...

A MICROORGANISM HAVING ENHANCED PRODUCTIVITY OF LACTIC ACID AND A PROCESS FOR PRODUCING LACTIC ACID USING THE SAME

The present invention relates to sp. capable of producing lactic acid with a decreased activity of pyruvate decarboxylase (PDC) and increased activities of aldehyde dehydrogenase (ALD) and acetyl-CoA synthetase (ACS), and a method of producing lactic acid from the culture medium obtained by culturing the microorganism. 1Saccharomyces cerevisiae. An isolated microorganism having enhanced productivity of lactic acid , wherein the microorganism is modified so that:a) the activity of pyruvate decarboxylase (PDC) of the microorganism is decreased compared to that of a non-modified lactic acid-producing strain; andb) the activities of aldehyde dehydrogenase (ALD) and acetyl-CoA synthetase (ACS) of the microorganism are enhanced compared to that of the non-modified lactic acid-producing strain.2. The microorganism according to claim 1 , wherein the pyruvate decarboxylase is at least one selected from the group consisting of PDC 1 claim 1 , PDC5 claim 1 , and PDC6.3. The microorganism according to claim 2 , wherein the microorganism is modified to:i) inactivate PDC1 activity and decrease PDC5 activity; orii) decrease PDC1 activity and inactivate PDC 5 activity.4. The microorganism according to claim 1 , wherein the aldehyde dehydrogenase is at least one selected from the group consisting of ALD2 and ALD3 claim 1 , and the acetyl-CoA synthetase is ACS1.5. The microorganism according to claim 1 , wherein alcohol dehydrogenase (ADH) is further inactivated.6. The microorganism according to claim 1 , wherein D-lactic acid dehydrogenase (DLD) is further inactivated.7. A method for producing lactic acid comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'a) culturing the microorganism according to in the culture medium; and'}b) recovering lactic acid from the culture medium or the microorganism in step a).8. A method for producing lactic acid comprising:{'claim-ref': {'@idref': 'CLM-00002', 'claim 2'}, 'a) culturing the microorganism according to in the culture medium; ...

Recombinant Cyanobacterial Cell For Contamination Control In A Cyanobacterial Culture Producing A Chemical Compound Of Interest

A non-naturally occurring cyanobacterial cell for the production of a chemical compound of interest is provided which contains at least one genetic modification that alters expression of at least one phosphate uptake regulating gene encoding a protein involved in regulation of phosphate metabolism, so that cellular uptake and/or intracellular storage of a phosphate compound is increased in comparison to a native form of the cyanobacterial cell. Related methods and uses involving the cyanobacterial cell are also provided. 2. The recombinant cyanobacterial cell of claim 1 , wherein the protein involved in regulation of phosphate metabolism represses expression and/or activity of the at least one phosphate transport protein in the cyanobacterial cell in its native form.3. The recombinant cyanobacterial cell of claim 1 , wherein the genetic modification causes constitutive expression and/or activity of the at least one phosphate transport protein.4. The recombinant cyanobacterial cell of claim 1 , further comprising a phosphate transport complex that comprises the at least one phosphate transport protein and additional proteins.5. The recombinant cyanobacterial cell of claim 1 , wherein the at least one phosphate transport protein and/or additional protein is selected from the group consisting of PstS claim 1 , PstC claim 1 , PstA claim 1 , PstB claim 1 , and combinations thereof.6. The recombinant cyanobacterial cell of claim 1 , wherein the at least one phosphate transport protein comprises a phosphate-binding protein capable of binding the phosphate compound.7. The recombinant cyanobacterial cell of claim 1 , wherein the at least one phosphate transport protein comprises an amino acid sequence having at least 16 identical residues without gaps to a consensus sequence VNYQSVGSGAGLRQFIXGTVDFAGSDLPL (SEQ ID NO: 41) claim 1 , wherein X is any one of the 20 natural amino acids.8. The recombinant cyanobacterial cell of claim 1 , wherein the genetic modification comprises a ...

MICROORGANISM HAVING ENHANCED PRODUCTIVITY OF LACTIC ACID AND A PROCESS FOR PRODUCING LACTIC ACID USING THE SAME

The present invention relates to sp. capable of producing lactic acid with a decreased activity of pyruvate decarboxylase (PDC) and increased activities of aldehyde dehydrogenase (ALD) and acetyl-CoA synthetase (ACS), and a method of producing lactic acid from the culture medium obtained by culturing the microorganism. 1Saccharomyces cerevisiae. An isolated microorganism having enhanced productivity of lactic acid , wherein the microorganism is modified so that:a) the activity of pyruvate decarboxylase (PDC) of the microorganism is decreased compared to that of a non-modified lactic acid-producing strain; andb) the activities of aldehyde dehydrogenase (ALD) and acetyl-CoA synthetase (ACS) of the microorganism are enhanced compared to that of the non-modified lactic acid-producing strain.2. The microorganism according to claim 1 , wherein the pyruvate decarboxylase is at least one selected from the group consisting of PDC1 claim 1 , PDC5 claim 1 , and PDC6.3. The microorganism according to claim 2 , wherein the microorganism is modified to:i) inactivate PDC1 activity and decrease PDC5 activity; orii) decrease PDC 1 activity and inactivate PDC 5 activity.4. The microorganism according to claim 1 , wherein the aldehyde dehydrogenase is at least one selected from the group consisting of ALD2 and ALD3 claim 1 , and the acetyl-CoA synthetase is ACS1.5. The microorganism according to claim 1 , wherein alcohol dehydrogenase (ADH) is further inactivated.6. The microorganism according to claim 1 , wherein D-lactic acid dehydrogenase (DLD) is further inactivated.7. A method for producing lactic acid comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'a) culturing the microorganism according to in the culture medium; and'}b) recovering lactic acid from the culture medium or the microorganism in step a).8. A method for producing lactic acid comprising:{'claim-ref': {'@idref': 'CLM-00002', 'claims 2'}, 'a) culturing the microorganism according to in the culture medium; ...

MICROORGANISMS AND METHODS FOR PRODUCTION OF SPECIFIC LENGTH FATTY ALCOHOLS AND RELATED COMPOUNDS

The invention provides non-naturally occurring microbial organisms containing a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length. Also provided are non-naturally occurring microbial organisms having a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms further include an acetyl-CoA pathway. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde or a fatty acid. 2. The non-naturally occurring microbial organism of claim 1 , wherein Ris Clinear alkyl claim 1 , Clinear alkyl claim 1 , Clinear alkyl claim 1 , C claim 1 , linear alkyl claim 1 , Clinear alkyl or Clinear alkyl.3. (canceled)4. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises two claim 1 , three claim 1 , or four exogenous nucleic acids each encoding an enzyme of said MD-FAE cycle claim 1 , or wherein said microbial organism comprises two claim 1 , three claim 1 , or four exogenous nucleic acids each encoding an enzyme of said termination pathway claim 1 , or wherein said microbial organism comprises exogenous nucleic acids encoding each of the enzymes of at least one of the pathways selected from (1)-(13).58-. (canceled)10. (canceled)11. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism further comprises an acetyl-CoA pathway and at least one exogenous nucleic acid encoding an acetyl-CoA pathway enzyme expressed in a sufficient amount to produce acetyl-CoA claim 1 , wherein said acetyl-CoA pathway comprises a pathway selected from:{'b': 2', '2, '(1) A and B;'}{'b': 2', '2', '2, '(2) A, C, and D;'}{'b': '2 ...

YEAST ORGANISM PRODUCING ISOBUTANOL AT A HIGH YIELD

The present invention provides recombinant microorganisms comprising an isobutanol producing metabolic pathway and methods of using said recombinant microorganisms to produce isobutanol. In various aspects of the invention, the recombinant microorganisms may comprise a modification resulting in the reduction of pyruvate decarboxylase and/or glycerol-3-phosphate dehydrogenase activity. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms. 1. A method of producing isobutanol , comprising:a) providing a recombinant microorganism comprising an isobutanol producing metabolic pathway, wherein the recombinant microorganism has been engineered to contain one or more modifications in a transcriptional regulator of a PDC gene;b) cultivating the microorganism in a culture medium containing a feedstock providing the carbon source, until a recoverable quantity of the isobutanol is produced; andc) recovering the isobutanol.2. The method of claim 1 , wherein the microorganism comprises an isobutanol producing metabolic pathway comprising the following substrate to product conversions:(i) pyruvate to acetolactate;(ii) acetolactate to 2,3-dihydroxyisovalerate;(iii) 2,3-dihydroxyisovalerate to α-ketoisovalerate;(iv) α-ketoisovalerate to isobutyraldehyde; and(v) isobutyraldehyde to isobutanol.3. The method of claim 1 , wherein the microorganism expresses(a) an acetolactate synthase to catalyze the conversion of pyruvate to acetolactate;(b) a ketol-acid reductoisomerase to catalyze the conversion of acetolactate to 2,3-dihydroxyisovalerate;(c) a dihydroxyacid dehydratase to catalyze the conversion of 2,3-dihydroxyisovalerate to α-ketoisovalerate;(d) an α-ketoisovalerate decarboxylase to catalyze the ...

METHODS FOR EFFICIENT PRODUCTION OF POLYUNSATURATED FATTY ACIDS(PUFA) IN RHODOSPORIDIUM AND RHODOTORULA SPECIES

The present invention relates to the field of fungal biotechnology, more particularly to genetic engineering methods for the production of polyunsaturated fatty acids (PUFA) in fungal hosts selected from and genera. The present invention further relates to a modified fungal host cell having reduced native aldehyde dehydrogenase (ALD 1) enzyme activity, and methods for producing omega-3 and omega-6 fatty acids and triacylglycerides, by growing said fungal host cell under suitable conditions. 1RhodosporidiumRhodotorula. A fungal host cell wherein the total fatty acids is composed of increased level of polyunsaturated fatty acids (PUFA) , in particular alpha-linolenic acid (ALA) or gamma-linolenic acid (GLA) , at a level of at least 9% , preferably more than 24% , most preferably more than 49% in the fungal host cell , wherein the genome of the fungal host cell has been modified such that the fungal host cell has reduced native aldehyde dehydrogenase (ALD1) enzyme activity compared to a fungal host cell having an unmodified genome , and wherein the fungal host is a species of the genera or the genera.2. The fungal host cell of claim 1 , wherein the native ALD1 is selected from the group consisting of:(a) ALD1 having the amino acid sequence set forth in SEQ ID NO:3; and(b) an ALD1 having at least 75% identity or at least 85% identity or at least 95% identity to the amino acid sequence set forth in SEQ ID NO:3.3. The fungal host cell of claim 2 , wherein the native ALD1 is encoded by a nucleic acid selected from the group consisting of:(a) a nucleic acid having the nucleotide sequence set forth in SEQ ID NO: 1;(b) a nucleic acid having the nucleotide sequence set forth in SEQ ID NO:2;(c) a nucleic acid having at least 75% identity or at least 85% identity or at least 95% identity to the nucleic acid of (a); and(d) a nucleic acid having at least 75% identity or at least 85% identity or at least 95% identity to the nucleic acid of (b).4. The fungal host cell of claim 1 , ...

MICROORGANISM HAVING ENHANCED PRODUCTIVITY OF LACTIC ACID AND A PROCESS FOR PRODUCING LACTIC ACID USING THE SAME

The present invention relates to sp. capable of producing lactic acid with a decreased activity of pyruvate decarboxylase (PDC) and increased activities of aldehyde dehydrogenase (ALD) and acetyl-CoA synthetase (ACS), and a method of producing lactic acid from the culture medium obtained by culturing the microorganism. 1Saccharomyces cerevisiae. An isolated microorganism having enhanced productivity of lactic acid , wherein the microorganism is modified so that:a) the activity of pyruvate decarboxylase (PDC) of the microorganism is decreased compared to that of a non-modified lactic acid-producing strain; andb) the activities of aldehyde dehydrogenase (ALD) and acetyl-CoA synthetase (ACS) of the microorganism are enhanced compared to that of the non-modified lactic acid-producing strain.2. The microorganism according to claim 1 , wherein the pyruvate decarboxylase is at least one selected from the group consisting of PDC1 claim 1 , PDC5 claim 1 , and PDC6.3. The microorganism according to claim 2 , wherein the microorganism is modified to:i) inactivate PDC1 activity and decrease PDC5 activity; orii) decrease PDC1 activity and inactivate PDC5 activity.4. The microorganism according to claim 1 , wherein the aldehyde dehydrogenase is at least one selected from the group consisting of ALD2 and ALD3 claim 1 , and the acetyl-CoA synthetase is ACS1.5. The microorganism according to claim 1 , wherein alcohol dehydrogenase (ADH) is further inactivated.6. The microorganism according to claim 1 , wherein D-lactic acid dehydrogenase (DLD) is further inactivated.7. A method for producing lactic acid comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'a) culturing the microorganism according to in the culture medium; and'}b) recovering lactic acid from the culture medium or the microorganism in step a).8. A method for producing lactic acid comprising:{'claim-ref': {'@idref': 'CLM-00002', 'claim 2'}, 'a) culturing the microorganism according to in the culture medium; and ...

FERMENTATIVE PRODUCTION OF ALCOHOLS

The invention relates to the development of microorganisms capable of producing fermentation products via an engineered pathway in the microorganisms. The invention also relates to microorganisms with improved cell viability and methods to improve cell viability and cell productivity of a microorganism. 1. A recombinant host cell comprising a modified yeast activator protein and/or yeast activator protein activity , wherein the host cell produces an alcohol.2. The recombinant host cell of claim 1 , wherein the recombinant host cell comprises a modification in a polynucleotide encoding a polypeptide having yeast activator protein activity.3. (canceled)4. The recombinant host cell of claim 2 , wherein the polypeptide having yeast activator protein activity is Yap1 claim 2 , Yap2 claim 2 , Yap3 claim 2 , Yap4 claim 2 , Yap5 claim 2 , Yap6 claim 2 , Yap7 claim 2 , or Yap8.5. The recombinant host cell of claim 2 , wherein the recombinant host cell comprises an amino acid substitution at residues 80 claim 2 , 132 claim 2 , 168 claim 2 , 237 claim 2 , 254 claim 2 , 257 claim 2 , 297 claim 2 , 302 claim 2 , 367 claim 2 , 404 claim 2 , 411 claim 2 , 444 claim 2 , 487 claim 2 , 498 claim 2 , 499 claim 2 , 548 claim 2 , 584 claim 2 , 617 claim 2 , and/or 636.6. (canceled)7. The recombinant host cell of claim 1 , wherein the recombinant host cell comprises one or more sequences selected from SEQ ID NO: 2 claim 1 , 3 claim 1 , 4 claim 1 , 5 claim 1 , 6 claim 1 , 7 claim 1 , 8 claim 1 , 9 claim 1 , 10 claim 1 , 11 claim 1 , 12 claim 1 , 13 claim 1 , 14 claim 1 , 15 claim 1 , 16 claim 1 , 17 claim 1 , 18 claim 1 , 19 claim 1 , 20 claim 1 , 21 claim 1 , 22 claim 1 , 23 claim 1 , 24 claim 1 , 25 claim 1 , 26 claim 1 , and 27.8. The recombinant host cell of claim 2 , wherein the recombinant host cell comprises one or more amino acid insertions.9. The recombinant host cell of claim 8 , wherein the recombinant host cell comprises one or more amino acid insertions of tripeptide repeat ...

Yeast Organism Producing Isobutanol at a High Yield

The present invention provides recombinant microorganisms comprising an isobutanol producing metabolic pathway and methods of using said recombinant microorganisms to produce isobutanol. In various aspects of the invention, the recombinant microorganisms may comprise a modification resulting in the reduction of pyruvate decarboxylase and/or glycerol-3-phosphate dehydrogenase activity. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the , Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms. 2. The recombinant yeast microorganism of claim 1 , wherein all endogenous PDC genes and all endogenous GPD genes are disrupted claim 1 , mutated claim 1 , or deleted.3. The recombinant yeast microorganism of claim 1 , wherein said one or more endogenous PDC genes is selected from the group consisting of PDC1 claim 1 , PDC2 claim 1 , PDC5 claim 1 , and PDC6.4. The recombinant yeast microorganism of claim 1 , wherein said one or more endogenous GPD genes is selected from the group consisting of GPD1 and GPD2.5. The recombinant yeast microorganism of claim 1 , wherein said ALS is a cytosolically-localized ALS.6Lactococcus lactis. The recombinant yeast microorganism of claim 5 , wherein said cytosolically-localized ALS is encoded by the alsS gene.7Bacillus subtilis. The recombinant yeast microorganism of claim 5 , wherein said cytosolically-localized ALS is encoded by the alsS gene.8. The recombinant yeast microorganism of claim 1 , wherein said recombinant yeast microorganism has been engineered to disrupt or delete an endogenous pyruvate dehydrogenase (PDH) gene.9. The recombinant yeast microorganism of claim 1 , wherein said recombinant yeast microorganism further expresses a heterologous gene encoding a ketol-acid reductoisomerase.10. The recombinant ...

Engineering of Acetyl-CoA Metabolism in Yeast

The invention relates to engineering of acetyl-CoA metabolism in yeast and in particular to production of acetyl-CoA in a non-ethanol producing yeast lacking endogenous gene(s) encoding pyruvate decarboxylase and comprising a heterologous pathway for synthesis of cytosolic acetyl-CoA. 122-. (canceled)23. A yeast comprising at least one heterologous pathway for synthesis of cytosolic acetyl-Coenzyme A (CoA) , the at least one heterologous pathway comprising at least one heterologous gene encoding a respective enzyme involved in synthesis of acetyl-CoA , with the proviso that the at least one heterologous gene is not a heterologous gene encoding a pyruvate formate lyase , wherein the at least one heterologous gene comprises:heterologous genes encoding a cytosolic pyruvate dehydrogenase complex; andheterologous genes encoding respective enzymes involved in attachment and activation of lipoyl groups to said cytosolic pyruvate dehydrogenase complex.24Escherichia coliSaccharomyces cerevisiaeAzotobacter vinelandiiEnterococcus faecalis. The yeast according to claim 23 , wherein the heterologous genes encoding said cytosolic pyruvate dehydrogenase complex are selected from the group consisting of genes encoding an cytosolic pyruvate dehydrogenase complex claim 23 , genes encoding a pyruvate dehydrogenase complex but lacking mitochondrial target signal (MTS) claim 23 , genes encoding an pyruvate dehydrogenase complex claim 23 , and genes encoding an pyruvate dehydrogenase complex; and{'i': Escherichia coli', 'Saccharomyces cerevisiae, 'the heterologous genes encoding respective enzymes involved in attachment and activation of lipoyl groups are selected from the group consisting of genes encoding lipoic acid synthetase and lipoic acid synthetase and/or lipoate-protein ligase, and genes encoding LIP2, LIP3, LIP5 and GCV3 but lacking MTS.'}25. The yeast according to claim 23 , wherein the yeast lacks any endogenous gene encoding pyruvate decarboxylase or comprises a disrupted ...

BACTERIA ENGINEERED TO TREAT DISORDERS INVOLVING THE CATABOLISM OF A BRANCHED CHAIN AMINO ACID

The present disclosure provides recombinant bacterial cells that have been engineered with genetic circuitry which allow the recombinant bacterial cells to sense a patient's internal environment and respond by turning an engineered metabolic pathway on or off. When turned on, the recombinant bacterial cells complete all of the steps in a metabolic pathway to achieve a therapeutic effect in a host subject. These recombinant bacterial cells are designed to drive therapeutic effects throughout the body of a host from a point of origin of the microbiome. Specifically, the present disclosure provides recombinant bacterial cells comprising a heterologous gene encoding a branched chain amino acid catabolism enzyme. The disclosure further provides pharmaceutical compositions comprising the recombinant bacteria, and methods for treating disorders involving the catabolism of branched chain amino acids using the pharmaceutical compositions disclosed herein. 1. A bacterium comprising gene sequence(s) encoding one or more branched chain amino acid catabolism enzyme(s) operably linked to a directly or indirectly inducible promoter that is not associated with the branched chain amino acid catabolism enzyme gene in nature.2. The bacterium of claim 1 , wherein the bacterium further comprises gene sequence(s) encoding one or more transporter(s) of a branched chain amino acid operably linked to a promoter that is not associated with the transporter gene in nature.3. The bacterium of claim 2 , wherein the promoter is a directly or indirectly inducible promoter.4. The bacterium of or claim 2 , wherein the bacterium further comprises a genetic modification that reduces export of a branched chain amino acid from the bacterium.5. The bacterium of or claim 2 , wherein the bacterium further comprises a genetic modification that reduces endogenous biosynthesis of a branched chain amino acid in the bacterium.6. The bacterium of or claim 2 , wherein the bacterium further comprises gene sequence ...

RECOMBINANT BACTERIA AND THE USES THEREOF FOR PRODUCING ETHANOL

The present invention relates to recombinant bacteria and the uses thereof, particularly for the production of ethanol. The invention also relates to methods for the production of such bacteria, as well as to nucleic acid constructs suitable for such production. The invention specifically relates to bacteria lacking a functional LDH gene and/or containing a recombinant nucleic acid encoding a PDC and ADH. The bacteria of this invention may be produced from any stress-resistant bacteria. 1Deinococcus. A recombinant bacterium comprising a recombinant nucleic acid construct encoding a pyruvate decarboxylase (PDC) and an alcohol dehydrogenase (ADH) , wherein the PDC and ADH coding sequences are placed in an operon in the recombinant nucleic acid construct , and wherein said recombinant nucleic acid construct is integrated into the genome of the bacterium.2. The bacterium of claim 1 , wherein said bacterium has a modified genome containing an at least partially inactive lactate dehydrogenase (LDH) gene.3. The bacterium of claim 2 , wherein the LDH gene is deleted all or in part and does not encode a functional protein.4. The bacterium of claim 2 , wherein the LDH gene has been at least partially inactivated by homologous recombination claim 2 , gene replacement or targeted mutagenesis.5. The bacterium of claim 1 , wherein the genome of said bacterium lacks at least 589 consecutive nucleotides of the LDH gene.6DeinococcusD. radiodurans, D. geothermalis, D murrayi, D. cellulosilyticusD. deserti.. The bacterium of claim 1 , wherein said bacterium is selected from or7Deinococcus. The bacterium of claim 1 , wherein said bacterium is a thermophilic bacterium.8. A method of producing a biofuel comprising cultivating a bacterium of in the presence of an appropriate substrate claim 1 , and collecting the biofuel.9. The method of claim 8 , wherein said cultivating is performed at a temperature of about 40° C. or more claim 8 , under acid pH conditions claim 8 , and/or is at least ...

Metabolically Enhanced Cyanobacterial Cell for the Production of Ethanol

A metabolically enhanced cyanobacterial cell for the production of ethanol is provided. The metabolically enhanced cyanobacterial cell for the production of ethanol comprises at least one recombinant gene encoding a pyruvate decarboxylase enzyme (Pdc) converting pyruvate to acetaldehyde, and at least one recombinant gene encoding a first Zn dependent alcohol dehydrogenase enzyme (Adh) converting acetaldehyde to ethanol. The invention also provides a method for producing the metabolically enhanced , a method for producing ethanol with the metabolically enhanced , and a method for screening of alcohol dehydrogenase enzyme expressing cyanobacterial strains for the presence of NADPH-dependent native alcohol dehydrogenase enzymes. 1. A method for producing ethanol in a cyanobacterial cell , comprising: i. a recombinant gene encoding a pyruvate decarboxylase enzyme (Pdc) converting pyruvate to acetaldehyde, and', {'sup': 2+', '2+, 'ii. a recombinant gene encoding a Zn dependent alcohol dehydrogenase enzyme (Adh) converting acetaldehyde to ethanol, wherein the amino acid sequence of said Zn dependent alcohol dehydrogenase enzyme (Adh) is at least 95% identical to an Adh amino acid sequence selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 8;'}], 'a. providing a cyanobacterial cell comprisingb. culturing said cyanobacterial cell in a growth medium under the exposure of light, the cyanobacterial cell producing ethanol while being cultured; andc. retrieving the ethanol from the cyanobacterial cell, the growth medium, and/or a headspace above the growth medium.2. The method of claim 1 , wherein the Adh amino acid sequence is at least 95% identical to a sequence selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 3.3. The method of claim of claim 2 , wherein:a. the Michaelis constant Km for NADPH of the alcohol dehydrogenase enzyme is lower than the Michaelis constant Km for NADH of the ...

Metabolically Enhanced Cyanobacterial Cell for the Production of Ethanol

A metabolically enhanced cyanobacterial cell for the production of ethanol is provided. The metabolically enhanced cyanobacterial cell for the production of ethanol comprises at least one recombinant gene encoding a pyruvate decarboxylase enzyme (Pdc) converting pyruvate to acetaldehyde, and at least one recombinant gene encoding a first Zn 2+ dependent alcohol dehydrogenase enzyme (Adh) converting acetaldehyde to ethanol. The invention also provides a method for producing the metabolically enhanced cyanobacterium , a method for producing ethanol with the metabolically enhanced cyanobacterium , and a method for screening of alcohol dehydrogenase enzyme expressing cyanobacterial strains for the presence of NADPH-dependent native alcohol dehydrogenase enzymes.

BACTERIAL COCULTURES EXPRESSING A BACTERIOCIN SYSTEM

The present disclosure concerns a co-culture of bacterial cells for making a fermented product from a biomass. The co-culture comprising a first recombinant lactic acid bacteria (LAB) cell expressing at least one bacteriocin and a second recombinant lactic acid bacteria (LAB) cell capable of converting, at least in part, the biomass into the fermented product. The second recombinant LAB cell is immune to the bacteriocin produced by the first recombinant LAB cell. The co-culture can be used, optionally in combination with a yeast host cell, to make a fermented product. The present disclosure also provides processes for making the fermented product by using the co-culture as wells kits and media comprising the co-culture. 1. A co-culture of lactic acid bacterial cells for making a fermented product from a biomass , the co-culture comprising: one or more first heterologous nucleic acid molecule encoding one or more polypeptide for converting, at least in part, the biomass into the fermented product;', 'optionally one or more second heterologous nucleic acid molecules encoding the at least one bacteriocin and/or the further bacteriocin and one or more polypeptide for conferring immunity against the at least one bacteriocin and/or the further bacteriocin; and, 'a first recombinant lactic acid bacteria (LAB) cell expressing at least one bacteriocin and optionally a further bacteriocin, wherein the first recombinant LAB cell comprisesa second recombinant LAB cell capable of converting, at least in part, the biomass into the fermented product, wherein the second recombinant LAB comprises:one or more third heterologous nucleic acid molecule encoding one or more polypeptide for converting, at least in part, the biomass into the fermented product; andone or more fourth heterologous nucleic acid molecule encoding one or more polypeptide for conferring immunity against the at least one bacteriocin and/or the further bacteriocin expressed by the first recombinant LAB cell.2. The ...

Construction of a Lactobacillus Casei Ethanologen

An engineered bacterium for producing ethanol from one or more carbohydrates is disclosed. The bacterium can be made by (a) inactivating within a bacterium one or more endogenous genes encoding a lactate dehydrogenase; or (b) introducing into a bacterium one or more exogenous genes encoding a pyruvate decarboxylase and one or more exogenous genes encoding an alcohol dehydrogenase II; or (c) performing both steps (a) and (b). The resulting engineered bacterium produces significantly more ethanol than the wild-type bacterium, and can be used in producing ethanol from a substrate such as biomass that includes carbohydrates. 1. A method of making ethanol comprising:{'i': Lactobacillus casei', 'L. casei, 'culturing on a substrate comprising a carbohydrate an engineered bacterium comprising a bacterium that includes one or more exogenous genes encoding a pyruvate decarboxylase and one or more exogenous genes encoding an alcohol dehydrogenase II, each of which is operably linked to a promoter that is highly expressed in the stationary phase, whereby the engineered bacterium produces a composition comprising ethanol.'}2. The method of claim 1 , wherein the engineered bacterium further includes one or more gene deletion mutations of one or more endogenous genes encoding a lactate dehydrogenase.3. The engineered bacterium of claim 1 , wherein the engineered bacterium further includes a gene deletion mutation of an endogenous gene encoding D-hydroxyisocaproate dehydrogenase.5. The method of claim 2 , wherein the gene deletion mutations comprise Δ L-lactate dehydrogenase 1 (ΔL-ldh1).5. The method of claim 2 , wherein the gene deletion mutations comprise Δ L-lactate dehydrogenase 2 (ΔL-ldh2).6. The method of claim 1 , wherein the engineered bacterium includes the gene deletion mutations Δ D-lactate dehydrogenase (ΔD-ldh) or Δ D-hydroxyisocaproate dehydrogenase (ΔD-hic).7Zymomonas mobilisZymomonas mobilis. The method of claim 1 , wherein the exogenous gene encoding a pyruvate ...

LYASE AND METHOD FOR ASYMMETRIC SYNTHESIS OF (S)-PHENYLACETYLCARBINOL

A lyase has an amino acid sequence selected from SEQ ID NOs: 1, 2 and 3, wherein the amino acid isoleucine in position no. 468 in the protein ApPDC-E469G, which is modified with respect to the wild type from is replaced by an amino acid which occupies less space than isoleucine. 1. A lyase ,comprising an amino acid sequence according to SEQ ID NO: 1, 2 or 3, wherein{'i': 'Acetobacter pasteurianus,', 'the amino acid isoleucine in position no. 468 in the protein ApPDC-E469G, which is modified with respect to the wild type from is replaced by an amino acid which occupies less space than isoleucine.'}2. (canceled)3. The lyase of claim 1 ,comprising an amino acid sequence according to SEQ ID NO: 23, whereinthe amino acid in position no. 543 is additionally replaced by an amino acid which occupies less space than tryptophan.4. (canceled)5. A deoxyribonucleic acid (DNA) molecule encoding the lyase of claim 1 , comprising a nucleotide sequence according to SEQ ID NO: 4 claim 1 ,wherein the DNA moleculeencodes a variant of the enzyme ApPDC-E469G in which the codon in position no. 1402-1404 is replaced by a codon which encodes an amino acid which occupies less space than isoleucine.6. A deoxyribonucleic acid (DNA) molecule encoding the lyase of claim 3 , comprising a nucleotide sequence according towherein the DNA moleculeencodes a variant of the enzyme ApPDC-E469G in which the codon in position no. 1402-1404 is replaced by a codon which encodes an amino acid which occupies less space than isoleucine and in which the codon in position no. 1627 to 1629 is additionally replaced by a codon which encodes an amino acid which occupies less space than tryptophan.7. (canceled)8. A vectorcomprising a nucleotide sequence according toSEQ ID NO: 4 or 48.9. The vector of claim 8 ,wherein the vectoris a plasmid.10. The vector of claim 9 ,whereinthe nucleotide sequence according to SEQ ID NO: 4 or 48 is ligated in an empty vector from the group pET-20b(+), pET-21a-d(+), pET-22b(+), pET-23a- ...

MICROORGANISMS AND METHODS FOR PRODUCTION OF SPECIFIC LENGTH FATTY ALCOHOLS AND RELATED COMPOUNDS

The invention provides non-naturally occurring microbial organisms containing a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length. Also provided are non-naturally occurring microbial organisms having a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms further include an acetyl-CoA pathway. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde or a fatty acid. 2. The non-naturally occurring microbial organism of claim 1 , wherein Ris Clinear alkyl.3. The non-naturally occurring microbial organism of claim 2 , wherein Ris Clinear alkyl claim 2 , Clinear alkyl claim 2 , C claim 2 , linear alkyl claim 2 , Clinear alkyl or Clinear alkyl.4. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises two claim 1 , three claim 1 , or four exogenous nucleic acids each encoding an enzyme of said MI-FAE cycle or said MD-FAE cycle.5. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises two claim 1 , three claim 1 , or four exogenous nucleic acids each encoding an enzyme of said termination pathway.6. The non-naturally occurring microbial organism of claim 3 , wherein said microbial organism comprises exogenous nucleic acids encoding each of the enzymes of at least one of the pathways selected from (1)-(13).7. The non-naturally occurring microbial organism of claim 1 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.8. The non-naturally occurring microbial organism of claim 1 , wherein said non-naturally occurring microbial organism is in a ...

CHEMOENZYMATIC SYNTHESIS OF PEPTIDE BETA-LACTONES AND BETA-HYDROXY ACIDS

Methods of producing peptide beta-lactones and beta-hydroxy acids are disclosed that include contacting a beta-hydroxy-alpha-amino acid, an aryl carrier protein (ObiD), and ATP with a non-ribosomal protein synthetase. A continuous flow reactor is disclosed that includes an elongate conduit with at least one region that includes a first region with a non-ribosomal protein synthetase immobilized to a substrate. The non-ribosomal protein synthetase of the continuous flow reactor is configured to contact a flow of a reaction mixture that includes a beta-hydroxy-alpha-amino acid and an aryl carrier protein. The non-ribosomal protein synthetase is further configured to release a peptide beta-lactone into the flow of the reaction mixture. 1. A method of producing a peptide beta-lactone , the method comprising contacting a beta-hydroxy-alpha-amino acid , a benzoic acid derivative , an aryl carrier protein , and ATP with a non-ribosomal protein synthetase , wherein:{'sub': '2', 'the beta-hydroxy-alpha-amino acid is selected from the group consisting of beta-OH-p-NO-homoPhe and beta-OH-homoPhe;'}the aryl carrier protein is selected from the group consisting of ObiD, a homolog of ObiD, recombinant ObiD, and any variation thereof comprising the amino acid sequence of SEQ ID NO: 1 or fragment thereof;the non-ribosomal protein synthetase is selected from the group consisting of ObiF, a homolog of ObiF, recombinant ObiF, and any variation thereof comprising the amino acid sequence of SEQ ID NO:2 or fragment thereof; andthe benzoic acid derivative is 2,3-dihydroxoybenzoic acid.3. The method of claim 1 , further comprising forming the beta-hydroxy-alpha-amino acid by contacting an aliphatic or aryl aldehyde or derivative thereof claim 1 , an amino acid claim 1 , and a pyridoxyl phosphate (PLP) cofactor with a serine hydroxymethyltransferase/threonine aldoloase claim 1 , wherein:the aliphatic or aryl aldehyde or derivative thereof is selected from the group consisting of aliphatic ...

HIGH YIELD ROUTE FOR THE PRODUCTION OF 1, 6-HEXANEDIOL

Provided herein are methods, compositions, and non-naturally occurring microbial organism for preparing compounds such as 1-butanol, butyric acid, succinic acid, 1,4-butanediol, 1-pentanol, pentanoic acid, glutaric acid, 1,5-pentanediol, 1-hexanol, hexanoic acid, adipic acid, 1,6-hexanediol, 6-hydroxy hexanoic acid, ε-Caprolactone, 6-amino-hexanoic acid, ε-Caprolactam, hexamethylenediamine, linear fatty acids and linear fatty alcohols that are between 7-25 carbons long, linear alkanes and linear α-alkenes that are between 6-24 carbons long, sebacic acid and dodecanedioic acid comprising: a) converting a Caldehyde and pyruvate to a Cβ-hydroxyketone intermediate through an aldol addition; and b) converting the Cβ-hydroxyketone intermediate to the compounds through enzymatic steps, or a combination of enzymatic and chemical steps. 168-. (canceled)69. A method , comprising an enzymatic step of converting a Caldehyde and pyruvate to a Cβ-hydroxyketone , wherein N is 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 or 22. This application is a continuation under 35 U.S.C. § 120 of International Application No. PCT/US2014/056175, filed Sep. 17, 2014, which in turn claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Application Nos. 61/878,996, filed Sep. 17, 2013, and 61/945,715, filed Feb. 27, 2014. All of the above-mentioned applications are incorporated herein by reference in their entirety.This disclosure relates generally to compositions and methods of preparation of industrially useful alcohols, amines, lactones, lactams, and acids, linear fatty acids and linear fatty alcohols that are between 7-25 carbons long, linear alkanes and linear co-alkenes that are between 6-24 carbons long.Throughout this disclosure, various publications, patents and published patent specifications are referenced by an identifying citation or by reference to an Arabic numeral. These publications, patents, and published patent ...

MICROORGANISMS THAT CO-CONSUME GLUCOSE WITH NON-GLUCOSE CARBOHYDRATES AND METHODS OF USE

Microorganisms that co-consume glucose with non-glucose carbohydrates, such as xylose, and methods of using same. The microorganisms comprise modifications that reduce or ablate the activity of a phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS) protein or modifications that reduce or ablate the activity of a phosphoglucose isomerase and a GntR. The PTS protein may be selected from an enzyme I (EI), an HPr, an FPr, and an enzyme II(EII). Additional modifications include reduction or ablation of the activity of a pyruvate formate lyase, a lactate dehydrogenase, and a fumarate reductase and inclusion of recombinant pyruvate decarboxylase and alcohol dehydrogenase genes. The microorganisms are particularly suited to co-consuming glucose and xylose in media containing these substrates and producing ethanol therefrom. 1. A recombinant microorganism comprising:a first genetic modification that reduces or ablates the activity of a phosphoglucose isomerases; anda second genetic modification selected from the group consisting of a genetic modification that reduces or ablates the activity of a GntR, a genetic modification that introduces a recombinant phosphogluconate dehydratase gene, a genetic modification that introduces a recombinant 2-keto-4-hydroxyglutarate aldolase gene, a genetic modification that introduces a recombinant 2-keto-3-deoxy-6-phosphogluconate aldolase gene, and a genetic modification that introduces a recombinant oxaloacetate decarboxylase gene.2. The recombinant microorganism of wherein the microorganism is a bacterium.3. (canceled)4. The recombinant microorganism of further comprising a genetic modification that reduces or ablates the activity of a pyruvate formate lyase.5. The recombinant microorganism of further comprising a genetic modification that reduces or ablates the activity of a lactate dehydrogenase.6. (canceled)7. The recombinant microorganism of further comprising a recombinant pyruvate decarboxylase gene and a ...

CELL-FREE PRODUCTION OF BUTANOL

Provided herein, in some aspects, are methods and compositions for producing large-scale quantities of butanol, including normal butanol (n-butanol), isobutanol, and 2-butanol using a cell-free system. 1. A method of producing a cell lysate for producing n-butanol , the method comprising:(a) culturing engineered cells that express at least one enzyme of a n-butanol biosynthetic pathway selected from the group consisting of: glucokinase, phosphoglucose isomerase, phosphofructokinase, fructose bisphosphate aldolase, triose phosphate isomerase, glyceraldehyde 3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, pyruvate dehydrogenase complex, acetyl-CoA acetyltransferase, hydroxybutyrl-CoA dehydrogenase, enoyl-CoA hydratase, crotonyl-CoA reductase, butyraldehyde dehydrogenase, and alcohol dehydrogenase, wherein the cells are cultured under conditions that result in expression of enzymes; and(b) lysing engineered cells cultured in step (a), thereby producing a cell lysate that comprises at least one enzyme of the n-butanol biosynthetic pathway.2. The method of claim 1 , wherein the engineered cells express at least 2 enzymes of the n-butanol biosynthetic pathway selected from the group consisting of: glucokinase claim 1 , phosphoglucose isomerase claim 1 , phosphofructokinase claim 1 , fructose bisphosphate aldolase claim 1 , triose phosphate isomerase claim 1 , glyceraldehyde 3-phosphate dehydrogenase claim 1 , phosphoglycerate kinase claim 1 , phosphoglycerate mutase claim 1 , enolase claim 1 , pyruvate kinase claim 1 , pyruvate dehydrogenase complex claim 1 , acetyl-CoA acetyltransferase claim 1 , hydroxybutyrl-CoA dehydrogenase claim 1 , enoyl-CoA hydratase claim 1 , crotonyl-CoA reductase claim 1 , butyraldehyde dehydrogenase claim 1 , and alcohol dehydrogenase.3. The method of claim 2 , wherein the engineered cells express 2 to 17 enzymes of the n-butanol biosynthetic pathway selected from the group consisting of: ...

Microorganism with knock-in at acetolactate decarboxylase gene locus

Provided herein is a genetically engineered microorganism comprising knock-in of DNA at an acetolactate decarboxylase gene locus. Replacement of the acetolactate decarboxylase gene with DNA encoding one or more native or nonnative enzymes confers certain advantages, including fermentation stability and increased production of native and nonnative products from gaseous substrates.

METHODS AND COMPOSITIONS FOR PRODUCTION OF ACETALDEHYDE

The present disclosure provides methods for producing acetaldehyde from renewable biological resources, for example, from a fermentable substrate, with the advantages of energy efficiency and ease of purification. Particular embodiments, feature the production of acetaldehyde from pyruvate, wherein the pyruvate is generated from various carbon sources (e.g., sugars) by pyruvate-producing microorganisms. The methods comprise culturing a pyruvate-producing microorganism in a culture medium under conditions such that pyruvate is excreted and accumulates extracellularly, resulting in a pyruvate-enriched medium. 119.-. (canceled)20. A microbial culture comprising:a) a culture medium;b) live cells of a microorganism that synthesizes and secretes pyruvate;c) extracellular pyruvate that has been synthesized and secreted into the culture medium of (a) by the microorganism of (b); andd) an extracellular pyruvate decarboxylating agent that is exogenous to the microorganism of (b).21Vibrio.. The culture of claim 20 , wherein the microorganism is a member of the genus22Escherichia coli.. The culture of claim 20 , wherein the microorganism is23. The culture of claim 20 , wherein the microorganism is a yeast.24. The culture of claim 20 , wherein the microorganism is a photosynthetic alga.25Chlamydomonas.. The culture of claim 24 , wherein the photosynthetic alga belongs to the genus26. The culture of claim 25 , wherein the photosynthetic alga has disruptions in the genes encoding pyruvate formate lyase and pyruvate decarboxylase.27. The culture of claim 20 , wherein the microorganism is genetically modified.28. The culture of claim 27 , wherein the microorganism has a blocked pyruvate-converting enzyme.29Escherichia coliSaccharomyces cerevisiae.. The culture of claim 27 , wherein the microorganism is selected from and30. The culture of claim 20 , wherein the pyruvate decarboxylating agent is pyruvate decarboxylase (PDC).31. The culture of claim 30 , further comprising a second ...

STRAIN EXPRESSING FRSA AND METHOD FOR PRODUCING ETHANOL USING SAME

The present application relates to a strain expressing the FrsA protein, and a method for producing ethanol using the same. The FrsA of the present application has a high PDC enzyme activity for a pyruvate, which is a substrate, and thus can be used in a process for producing ethanol. In addition, an FrsA mutant having improved stability in a host cell can be more effective in producing ethanol due to the increase in stability when the FrsA mutant is overexpressed together with IIA, compared with when using conventional -derived PDC 1. A cell transformed with a FrsA gene.2. The cell of claim 1 , wherein the gene is represented by SEQ ID Nos: 1 claim 1 , 5 or 7.3Escherichia coli, Corynebacterium glutamicum, Erwinia chrysanthemi, Zymomonas mobilis, KlebsiellaBacillus stearothrermophilus, KluveromycesPachysolen tanophilus, ClostridiumCandida shehataeSaccharomyces seravisiaePichia stipitis.. The cell of claim 2 , wherein the single cell organism is a bacterium or a yeast in which the bacteria is selected from the group consisting of spp. claim 2 , spp. claim 2 , spp. and ; and the yeast is or4Escherichia coliCorynebacterium glutamicum.. The cell of claim 3 , wherein the bacteria is claim 3 , or5. The cell of claim 1 , wherein the bacterium is further transformed with a alcohol dehydrogenase gene.6. The cell according to claim 1 , wherein the cell is further transformed with a IIAgene.7. The cell of claim 6 , wherein the IIAgene is represented by SEQ ID NO: 3.8. A FrsA protein represented by SEQ ID NO: 2 in which the amino acid residue 131 is substituted from cysteine to alanine claim 6 , or the amino acid residues from 2 to 19 are deleted and the amino acid residue 131 is substituted from cysteine to alanine.9. An isolated polynucleotide encoding the protein according to .10. The polynucleotide of claim 9 , wherein the polynucleotide is represented by SEQ ID NOs: 5 or 7.11. A vector comprising the polynucleotide according to .12. A microorganism comprising the vector ...

RECOMBINANT BACTERIA ENGINEERED TO TREAT DISEASES AND DISORDERS ASSOCIATED WITH AMINO ACID METABOLISM AND METHODS OF USE THEREOF

The present disclosure provides recombinant bacterial cells that have been engineered with genetic circuitry which allow the recombinant bacterial cells to sense a patient's internal environment and respond by turning an engineered metabolic pathway on or off. When turned on, the recombinant bacterial cells complete all of the steps in a metabolic pathway to achieve a therapeutic effect in a host subject. These recombinant bacterial cells are designed to drive therapeutic effects throughout the body of a host from a point of origin of the microbiome. Specifically, the present disclosure provides recombinant bacterial cells that comprise an amino acid catabolism enzyme for the treatment of diseases and disorders associated with amino acid metabolism, including cancer, in a subject. The disclosure further provides pharmaceutical compositions and methods of treating disorders associated with amino acid metabolism, such as cancer. 1. A recombinant bacterial cell comprising a heterologous gene sequence encoding an amino acid catabolism enzyme operably linked to a first promoter that is not associated with the gene encoding the amino acid catabolism enzyme in nature.2. (canceled)3. (canceled)4. The recombinant bacterial cell of claim 1 , wherein the first promoter is directly or indirectly induced by environmental conditions specific to the gut of a mammal.5. The recombinant bacterial cell of claim 4 , wherein the first promoter is directly or indirectly induced by low oxygen or anaerobic conditions.6. The recombinant bacterial cell of claim 5 , wherein the first promoter is an FNR responsive promoter.7. The recombinant bacterial cell of claim 1 , wherein the first promoter is a constitutive promoter.8. The recombinant bacterial cell of further comprising a heterologous gene encoding an amino acid transporter.9. The recombinant bacterial cell of wherein the heterologous gene encoding the amino acid transporter is operably linked to a second promoter that is not associated ...

ENZYMATIC METHOD FOR PRODUCING 2-HYDROXY-4-METHYLMERCAPTOBUTANOIC ACID (MHA)

The invention relates to an enzymatic method for producing 2-hydroxy-4-methylmercaptobutanoic acid from 3-methylthio-propanal (3-methylmercaptopropanal (MMP) or “methional”) and carbon dioxide. 1. A method for producing D- or L-2-hydroxy-4-methylmercaptobutanoic acid (MHA) , comprising reacting a mixture comprising: 3-(methylthio)-propanal (methional); carbon dioxide; a decarboxylase (EC 4.1.1); a corresponding cofactor of the decarboxylase; an alcohol dehydrogenase (EC 1.1.1); and NADH or NADPH , to form D- or L-2-hydroxy-4-methylmercaptobutanoic acid (MHA) or a salt thereof.2. The method of claim 1 , wherein the cofactor comprises thiamine pyrophosphate.3Saccharomyces cerevisiaeSaccharomyces cerevisiaeLactococcus lactis. The method of claim 1 , wherein the decarboxylase is selected from the group consisting of pyruvate decarboxylase Pdc1 claim 1 , which originates from claim 1 , phenylpyruvate decarboxylase Aro10 claim 1 , which originates from claim 1 , and branched chain decarboxylase KdcA claim 1 , which originates from claim 1 , as well as mutants and variants of these decarboxylase having decarboxylase activity.4. The method of claim 1 , wherein said method is for producing D-2-hydroxy-4-methylmercapto-butanoic acid (D-MHA) claim 1 , and wherein the alcohol dehydrogenase is a D-hydroxyisocaproate dehydrogenase.5Lactobacillus casei. The method of claim 4 , wherein the D-hydroxyisocaproate dehydrogenase is D-HicDH from or any of its mutants and variants having alcohol dehydrogenase activity.6. The method of claim 1 , where said method is for producing L-2-hydroxy-4-methylmercapto-butanoic acid (L-MHA) claim 1 , and wherein the alcohol dehydrogenase is a L-hydroxyisocaproate dehydrogenase.7Lactobacillus confusus. The method of claim 6 , wherein the L-hydroxyisocaproate dehydrogenase is L-HicDH from or any of its mutants and variants having alcohol dehydrogenase activity.8. The method of claim 1 , wherein the carbon dioxide is applied to the mixture at a pressure ...

Production of glucan polymers from alternate sucrose sources

Reaction solutions are disclosed herein comprising water, incompletely refined sucrose, and a glucosyltransferase enzyme that synthesizes insoluble poly alpha-1,3-glucan having at least 50% alpha-1,3 glycosidic linkages and a weight average degree of polymerization (DP w ) of at least 100. The yield of poly alpha-1,3-glucan by a reaction solution herein is at least 7% of the weight of sucrose that was converted in the reaction solution. Further disclosed are methods of producing poly alpha-1,3-glucan using incompletely refined sucrose, and poly alpha-1,3-glucan produced by these methods.

Fermentive Production of Isobutanol Using Highly Effective Ketol-Acid Reductoisomerase Enzymes

Ketol-acid reductoisomerase enzymes have been identified that provide high effectiveness in vivo as a step in an isobutanol biosynthetic pathway in bacteria and in yeast. These KARIs are members of a clade identified through molecular phylogenetic analysis called the SLSL Clade. 118-. (canceled)19. A recombinant yeast cell comprising at least one nucleic acid molecule encoding a polypeptide having ketol-acid reductoisomerase (KARI) activity wherein the polypeptide having KARI activity has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 28 , 36 , 38 , 40 , 56 , 60 , or 68 and wherein the yeast cell comprises:(i) at least one inactivated pyruvate decarboxylase (PDC) gene,(ii) a modification to reduce glycerol-3-phosphate dehydrogenase activity, and(iii) a heterologous polynucleotide encoding a polypeptide with phosphoketolase activity.20. The recombinant yeast cell of claim 19 , wherein the inactivated pyruvate decarboxylase (PDC) gene is PDC1 claim 19 , PDC5 claim 19 , or PDC6.21. The recombinant yeast cell of claim 19 , wherein the recombinant yeast cell further comprises at least one deletion claim 19 , mutation claim 19 , or substitution in an endogenous gene encoding a polypeptide affecting Fe—S cluster biosynthesis.22. The recombinant yeast cell of claim 19 , wherein the yeast cell expresses an engineered isobutanol biosynthetic pathway.23. The recombinant yeast cell of claim 22 , wherein the engineered isobutanol biosynthetic pathway comprises the following substrate to product conversions:(i) pyruvate to acetolactate catalyzed by acetolactate synthase,(ii) 2,3-dihydroxyisovalerate to a-ketoisovalerate catalyzed by dihydroxy-acid dehydratase, and(iii) α-ketoisovalerate to isobutyraldehyde catalyzed by branched-chain a-keto acid decarboxylase.24. The recombinant yeast cell of claim 23 , wherein the acetolactate synthase has an EC number 1.1.1.86.25. The recombinant yeast cell of claim 23 , wherein the dihydroxy-acid ...

ESCHERICHIA COLI ENGINEERED FOR ISOBUTYRALDEHYDE PRODUCTION

The present disclosure provides recombinant and other bacteria with reduced alcohol dehydrogenase and/or aldehyde reductase activity. The present disclosure further provides recombinant and other bacteria with reduced isobutyraldehyde reductase activity. Methods for the production and the uses of the recombinant bacteria are also provided. Specifically, recombinant bacteria further expressing 2-keto-acid decarboxylase can be used for producing higher aldehydes or other non-alcohol chemicals derived from aldehydes. 1E. coli. An comprising a mutation in each of:(a) adhE, yqhD, adhP, eutG, yiaY, and yjgB genes, wherein said mutation reduces alcohol dehydrogenase activity of products of said genes; or(b) adhE, yqhD, adhP, eutG, fucO, and yjgB genes, and wherein said mutation reduces isobutyraldehyde reductase activity of products of said genes.2E. coliE. coli. The of claim 1 , wherein said comprises a mutation in each of adhE claim 1 , yqhD claim 1 , adhP claim 1 , eutG claim 1 , yiaY claim 1 , and yjgB genes claim 1 , and wherein said mutation reduces alcohol dehydrogenase activity of products of said genes.3E. coliE. coli. The of claim 1 , wherein said comprises a mutation in each of adhE claim 1 , yqhD claim 1 , adhP claim 1 , eutG claim 1 , fucO claim 1 , and yjgB genes claim 1 , and wherein said mutation reduces isobutyraldehyde reductase activity of products of said genes.4E. coliE. coli. The of claim 1 , wherein said further comprises a 2-keto-acid decarboxylase.5E. coli. The of claim 4 , wherein said 2-keto-acid decarboxylase is Kivd.6E. coliE. coliE. coli.. The of claim 5 , wherein said produces an increased level of a 2-keto-acid as compared to wild type7E. coliE. coli. The of claim 6 , wherein said has elevated expression or activity of:a) acetohydroxy acid synthase or acetolactate synthase;b) acetohydroxy acid isomeroreductase; orc) dihydroxy-acid dehydratase;{'i': 'E. coli.', 'or any combination of the foregoing, as compared to wild type'}8E. coliE. coliE. ...

LACTIC ACID BACTERIA FOR THE PRODUCTION OF ETHANOL FROM BIOMASS MATERIAL

Lactic acid bacterial cultures, cell populations and articles of manufacture comprising same are disclosed for generating ethanol from lignocellulose. 1. A bacterial culture comprising a biomass composition and a population of lactic acid bacteria which comprises:(i) a first population of lactic acid bacteria which has been genetically modified to express a secreted cellulase;(ii) a second population of lactic acid bacteria which has been genetically modified to express a secreted xylanase, wherein the ratio of the first population:second population is selected such that the specific activity of cellulase:xylanase in the culture is greater than 4:1 or less than 1:4; and(iii) a third population of lactic acid bacteria which has been genetically modified to produce ethanol.2. A bacterial culture comprising a biomass composition and a population of lactic acid bacteria which comprises:(i) a first population of lactic acid bacteria which has been genetically modified to express a secreted cellulase;(ii) a second population of lactic acid bacteria which has been genetically modified to express a secreted xylanase, wherein the ratio of the first population:second population is selected such that the specific activity of cellulase:xylanase in the culture is greater than 4:1 or less than 1:4, wherein said first and/or said second population of lactic acid bacteria has been further genetically modified to produce ethanol.3. An article of manufacture comprising:(i) a first population of lactic acid bacteria which are genetically modified to express at least one fibrolytic enzyme; and(ii) a second population of lactic acid bacteria which are genetically modified to produce ethanol from C5 or C6 sugars.4. The article of manufacture of claim 3 , wherein said first population of lactic acid bacteria express a cellulase.5. The article of manufacture of claim 4 , further comprising a third population of lactic acid bacteria claim 4 , which are genetically modified to express a ...

EXPRESSION OF A HAP TRANSCRIPTIONAL COMPLEX SUBUNIT

The invention relates, for example, to recombinant yeast cells for differential gene expression during the propagation and production phases of a fermentation-based production process, as well as methods for using the same. 1. A recombinant yeast cell , comprising (a) a recombinant polynucleotide encoding a gene for a subunit of the HAP transcriptional complex; and (b) an engineered isobutanol biosynthetic pathway.2. The recombinant yeast cell of claim 1 , wherein the subunit is Hap2 claim 1 , Hap3 claim 1 , Hap4 or Hap5.3. The recombinant yeast cell of claim 1 , wherein the subunit comprises an amino acid sequence at least 80% claim 1 , at least 85% claim 1 , at least 90% claim 1 , at least 95% claim 1 , at least 96% claim 1 , at least 97% claim 1 , at least 98% claim 1 , or at least 99% identical to any of SEQ ID NOs: 2 claim 1 , 4 claim 1 , 6 claim 1 , or 8.4. The recombinant yeast cell of claim 1 , wherein the polynucleotide comprises a nucleic acid sequence at least 80% claim 1 , at least 85% claim 1 , at least 90% claim 1 , at least 95% claim 1 , at least 96% claim 1 , at least 97% claim 1 , at least 98% or at least 99% identical to any of SEQ ID NOs: 1 claim 1 , 3 claim 1 , 5 claim 1 , or 7.5. The recombinant yeast cell of claim 1 , wherein the gene is expressed during propagation phase of a fermentation-based production process.6. The recombinant yeast cell of claim 1 , wherein the gene is down-regulated or not expressed during production phase of a fermentation-based production process.7. The recombinant yeast cell of claim 1 , wherein the gene is operably linked to a conditional promoter.8. (canceled)9. The recombinant yeast cell of claim 7 , wherein the conditional promoter is ADH2 claim 7 , HXT5 or HXT7.10. The recombinant yeast cell of claim 7 , wherein the conditional promoter comprises a nucleic acid sequence at least 80% claim 7 , at least 85% claim 7 , at least 90% claim 7 , at least 95% claim 7 , at least 96% claim 7 , at least 97% claim 7 , at ...

YEAST CELL HAVING ACID TOLERANCE, METHOD OF PREPARING YEAST CELL AND USE THEREOF

Provided are a genetically engineered yeast cell having increased activity of SUL1, STR3, HXT7, ERR1, GRX8, MXR1, GRE1, MRK1, AAD10 or a combination thereof, compared to a parent cell, and also having acid tolerance, a method of preparing the same, and a method of producing lactate using the same. 1. A genetically engineered yeast cell comprising a genetic modification that increases the activity of SUL1 , STR3 , HXT7 , ERR1 , GRX8 , MXR1 , GRE1 , MRK1 , AAD10 , or a combination thereof , as compared to a parent cell thereof; and having increased acid tolerance as compared to a parent cell thereof.2. The genetically engineered yeast cell of claim 1 , in which expression of a gene encoding SUL1 claim 1 , STR3 claim 1 , HXT7 claim 1 , ERR1 claim 1 , GRX8 claim 1 , MXR1 claim 1 , GRE1 claim 1 , MRK1 claim 1 , AAD10 claim 1 , or a combination thereof is increased claim 1 , compared to a parent cell thereof.3. The genetically engineered yeast cell of claim 1 , comprising a genetic modification within an expression regulatory sequence of a gene encoding SUL1 claim 1 , STR3 claim 1 , HXT7 claim 1 , ERR1 claim 1 , GRX8 claim 1 , MXR1 claim 1 , GRE1 claim 1 , MRK1 claim 1 , AAD10 claim 1 , or a combination thereof.4. The genetically engineered yeast cell of claim 1 , wherein the amino acid sequence of SUL1 claim 1 , STR3 claim 1 , HXT7 claim 1 , ERR1 claim 1 , GRX8 claim 1 , MXR1 claim 1 , GRE1 claim 1 , MRK1 claim 1 , or AAD10 has at least 95% sequence identity to SEQ ID NO: 1 claim 1 , 3 claim 1 , 5 claim 1 , 7 claim 1 , 9 claim 1 , 11 claim 1 , 13 claim 1 , 15 claim 1 , and 17 claim 1 , respectively.5. The genetically engineered yeast cell of claim 1 , wherein the gene encoding SUL1 claim 1 , STR3 claim 1 , HXT7 claim 1 , ERR1 claim 1 , GRX8 claim 1 , MXR1 claim 1 , GRE1 claim 1 , MRK1 claim 1 , or AAD10 has at least 95% sequence identity to SEQ ID NO: 2 claim 1 , 4 claim 1 , 6 claim 1 , 8 claim 1 , 10 claim 1 , 12 claim 1 , 14 claim 1 , 16 claim 1 , or 18 claim 1 , ...

MICROORGANISM PRODUCING LACTIC ACID AND METHOD FOR PRODUCING LACTIC ACID USING SAME

The present application relates to a microorganism of the genus producing lactic acid and a method for preparing lactic acid using the same. More specifically, the present application relates to a microorganism of the genus producing lactic acid, wherein the microorganism is modified to weaken or inactivate the activity of pyruvate decarboxylase (PDC) compared to its endogenous activity, to introduce the activity of ATP-citrate lyase (ACL), and to enhance pyruvate biosynthetic pathway compared to its endogenous biosynthetic pathway, and a method for producing lactic acid using the microorganism. 1Saccharomyces. A microorganism of the genus producing lactic acid , wherein the microorganism is modified to have inactivated activity of pyruvate decarboxylase (PDC) compared to its endogenous activity , to introduce the activity of ATP-citrate lyase (ACL) , and to enhance pyruvate biosynthetic pathway compared to its endogenous biosynthetic pathway.2. The microorganism according to the claim 1 , wherein the enhancement of pyruvate biosynthetic pathway is achieved by enhancing the activity of phosphoenolpyruvate carboxykinase 1 (PCK1) claim 1 , pyruvate kinase 2 (PYK2) or the activities of both claim 1 , compared to their endogenous activities.3. The microorganism according to the claim 1 , wherein the enhancement of pyruvate biosynthetic pathway is achieved by enhancing the activity of malate dehydrogenase 2 (MDH2) claim 1 , cytosolic malic enzyme 1 (cytosolic MAE1) or the activities of both claim 1 , compared to their endogenous activities.4. The microorganism according to the claim 1 , wherein pyruvate decarboxylase is an enzyme represented by at least one amino acid sequence selected from the group consisting of amino acid sequences of SEQ ID NOS: 39 claim 1 , 41 claim 1 , and 43.5. The microorganism according to the claim 1 , wherein ATP-citrate lyase is an enzyme represented by an amino acid sequence of SEQ ID NO. 29.6. The microorganism according to the claim 2 , ...

Integration of a Polynucleotide Encoding a Polypeptide That Catalyzes Pyruvate to Acetolactate Conversion

The invention relates to recombinant host cells having at least one integrated polynucleotide encoding a polypeptide that catalyzes a step in a pyruvate-utilizing biosynthetic pathway, e.g., pyruvate to acetolactate conversion. The invention also relates to methods of increasing the biosynthetic production of isobutanol, 2,3-butanediol, 2-butanol or 2-butanone using such host cells. 147-. (canceled)48. A recombinant host cell comprising:{'i': Bacillus subtilis, Klebsiella pneumonia, Lactococcus lactis, Staphylococcus aureus, Listeria monocytogenes, Streptococcus mutans, Streptococcus thermophiles, Vibrio angustum,', 'Bacillus cereus;, '(a) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of pyruvate to acetolactate wherein the polypeptide is an acetolactate synthase from or'}(b) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of acetolactate to 2,3-dihydroxyisovalerate wherein the polypeptide is a ketol-acid reductoisomerase and the ketol-acid reductoisomerase has at least 95% identity to SEQ ID NO: 83;{'i': Escherichia coli, Bacillus subtilis, Methanococcus maripaludis,', 'Streptococcus mutans;, '(c) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of 2,3-dihydroxyisovalerate to α-ketoisovalerate wherein the polypeptide is a dihydroxyacid dehydratase from or and'}(d) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of α-ketoisovalerate to isobutyraldehyde wherein the polypeptide is a branched-chain α-keto acid decarboxylase and the branched-chain α-keto acid decarboxylase has at least 95% identity to SEQ ID NO: 247.49Achromobacter xylosoxidansBeijerinkia indica.. The recombinant host cell of further comprising a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion isobutyraldehyde to isobutanol wherein the polypeptide is an alcohol dehydrogenase from or50. ...

NOVEL PICHIA KUDRIAVZEVII NG7 MICROORGANISM AND USES THEREOF

The present invention relates to: a novel microorganism NG7 showing heat resistance and acid resistance; a composition, for producing organic acid or alcohol, which comprises the microorganism and a culture of the same; and a method, for producing an organic acid or alcohol, which comprises culturing the microorganism. 1Pichia kudriavzevii. A NG7 microorganism having acid resistance and heat resistance.2. The microorganism of claim 1 , wherein the microorganism is deposited under the Accession Number KCTC12840BP.3. The microorganism of claim 1 , wherein an activity of orotidine 5′-phosphate decarboxylase (URA3) is further inactivated.4. The microorganism of claim 1 , wherein an activity of pyruvate decarboxylase is weakened compared to its endogenous activity and an activity of lactate dehydrogenase is further introduced.5. The microorganism of claim 4 , wherein the microorganism is deposited under the Accession Number KCTC12841BP.6. The microorganism of claim 1 , wherein the microorganism has an ability to use glycerol as a carbon source.7. The microorganism of claim 3 , wherein the activity of pyruvate decarboxylase is inactivated and an activity of lactate dehydrogenase is further introduced.8. The microorganism of claim 7 , wherein the microorganism is deposited under the Accession Number KCTC12842BP.9. A composition for producing an organic acid or alcohol claim 1 , comprising the microorganism of or a culture thereof.10. The composition of claim 9 , wherein the composition further comprises glycerol.11. The composition of claim 9 , wherein the organic acid is lactic acid.12. A method for producing an organic acid or alcohol claim 1 , comprising culturing the microorganism of in medium.13. The method of claim 12 , wherein the organic acid is lactic acid.14. The method of claim 12 , wherein the medium comprises glycerol.15. The method of claim 12 , wherein the cultivation is performed in a condition without adding a neutralizing agent. The present invention ...

PROCESS FOR PRODUCING L-METHIONINE FROM METHIONAL

A method is useful for the biocatalytic synthesis of proteinogenic L-amino acids, such as L-alanine, L-valine. L-methionine. L-leucine, L-isoleucine or L-phenylalanine from a respective aldehyde and carbon dioxide. In particular, the method is useful for the biocatalytic synthesis of L-methionine from 3-methylthio-propanal (“methional”) and carbon dioxide. 1: A method for producing an L-amino acid , comprising: (a) at least one donor amino acid and an aminotransferase, and/or', '(b) NADH, ammonia and/or an ammonium salt and an amino acid dehydrogenase,, 'reacting a mixture comprising an aldehyde, carbon dioxide, a decarboxylase, a corresponding decarboxylase cofactor, and'}to form said L-amino acid or a salt thereof.2: The method as claimed in claim 1 , wherein the L-amino acid to be formed is at least one member selected from the group consisting of L-alanine claim 1 , L-valine claim 1 , L-methionine claim 1 , L-leucine claim 1 , L-isoleucine and L-phenylalanine.3: The method as claimed in claim 1 , wherein the L-amino acid to be formed is L-methionine and the aldehyde is 3-(methylthio)-propanal (methional).4: The method as claimed in claim 1 , wherein the corresponding decarboxylase cofactor comprises thiamine pyrophosphate.5Saccharomyces cerevisiaeSaccharomyces cerevisiaeLactococcus lactis: The method as claimed in claim 1 , wherein the decarboxylase is at least one member selected from the group consisting of pyruvate decarboxylase PDC1 claim 1 , which originates from claim 1 , phenylpyruvate decarboxylase ARO10 claim 1 , which originates from claim 1 , and branched chain decarboxylase KdcA claim 1 , which originates from claim 1 , as well as mutants and variants of these decarboxylases having decarboxylase activity.6E. coliSaccharomyces cerevisiae: The method as claimed in claim 1 , wherein the aminotransferase is present claim 1 , and is at least one member selected from the group consisting of methionine aminotransferase YbdL claim 1 , which originates from ...

Ethanol Production in Microorganisms

The present disclosure relates to methods and compositions for engineering photoautotrophic organisms to convert carbon dioxide and light into fatty acid esters and other molecules, including biofuels. The molecules are then secreted by the organism into a growth medium.

MODIFIED MICROORGANISMS AND METHODS FOR PRODUCTION OF USEFUL PRODUCTS

Non-naturally occurring microbial organisms and related methods, processes and materials are for microbial organisms that include a genetic modification which enhances production of 3-hydroxybutanal or a downstream product of 3-hydroxybutanal such as 1,3-butanediol from endogenous central metabolic intermediates such as acetyl CoA or pyruvate which are converted to acetaldehyde. Two molecules of acetaldehyde are condensed to form the 3-hydroxybutanal using an aldolase capable of accepting acetaldehyde as both the acceptor and donor in an aldol condensation. The aldolase may be a deoxyribose phosphate aldolase type enzyme, and is typically introduced into the organisms. Energetically favorable pathways produce 3-hydroxybutanal or downstream products thereof. 1. A non-naturally occurring microbial organism which includes a genetic modification in its genome which enhances production of 3-hydroxybutanal or a downstream product of 3-hydroxybutanal by the microbial organism from at least one endogenous central metabolic intermediate via a 3-hydroxybutanal synthetic pathway in which two molecules of acetaldehyde are condensed to form said 3-hydroxybutanal using an aldolase capable of accepting acetaldehyde as both the acceptor and donor in an aldol condensation.2. (canceled)3. A non-naturally occurring microbial organism as claimed in whereinthe genetic modification:(i) introduces a heterologous gene encoding an enzyme having an activity utilised in generation of acetaldehyde from one or more of the central metabolic intermediates;(ii) up-regulates at least one endogenous enzyme having an activity utilised in generation of acetaldehyde from one or more of the central metabolic intermediates; and/or(iii) down-regulates or inactivates an endogenous enzyme which utilises acetaldehyde as a substrate,thereby increasing production or availability to the aldolase of the acetaldehyde, thereby increasing production of the 3-hydroxybutanal from the aldolase, and/orwherein the ...

AMINO ACID-PRODUCING MICROORGANISMS AND METHODS OF MAKING AND USING

Modified or recombinant microorganisms are provided herein that can be used to produce one or more amino acids, including, for example, methionine or one or more methionine biosynthetic pathway-derived intermediates or one or more methionine-based products. 1. A method of synthesizing methionine or one or more methionine biosynthetic pathway-derived intermediates claim 41 , the method comprising culturing a methanotrophic recombinant bacterial strain of in a culture medium comprising:(a) methane, methanol, sugars, glycerol, or a combination thereof; and(b) a source of sulfur.221.-. (canceled)22. The method of claim 1 , further comprising harvesting methionine or a methionine biosynthetic pathway intermediate synthesized by the recombinant methanotrophic bacterial strain.23. (canceled)24. The method of claim 22 , wherein the methionine biosynthetic pathway intermediate is selected from the group consisting of: oxaloacetate claim 22 , aspartate claim 22 , homoserine claim 22 , O-succinyl-L-homoserine claim 22 , O-acetyl-L-homoserine claim 22 , cystathionine claim 22 , homocysteine claim 22 , and S-adenosyl-methionine.2537.-. (canceled)38. The method of claim 22 , further comprising adding the harvested methionine or the harvested methionine biosynthetic pathway intermediate to an animal feed or food product.3940.-. (canceled)41. A recombinant methanotrophic bacterial strain comprising heterologous nucleic acid encoding at least one protein and/or comprises nucleic acid encoding at least inactivated endogenous protein claim 22 , that is capable of producing L-methionine or a methionine biosynthetic pathway-derived intermediate in a culture medium under conditions sufficient to produce L-methionine or a methionine biosynthetic pathway-derived intermediate claim 22 , wherein the culture medium comprises:(a) methane, methanol, sugars, glycerol, or a combination thereof; and(b) a source of sulfur.4244.-. (canceled)45. The recombinant methanotrophic bacterial strain of ...

Mutant of Genus Rhizopus

The present invention provides a fungus of the genus having high productivity of an organic acid. The present invention also provides a mutant of the genus with reduced pyruvate decarboxylase activity. 1Rhizopus. A mutant of the genus wherein the mutant's pdc gene is deleted or inactivated , whereinthe pdc gene is at least one polynucleotide selected from the group consisting of:a polynucleotide consisting of the nucleotide sequence represented by SEQ ID NO: 1;a polynucleotide consisting of a nucleotide sequence having at least 80% identity to the nucleotides represented by SEQ ID NO: 1, and encoding a polypeptide which has pyruvate decarboxylase activity;a polynucleotide consisting of a nucleotide sequence having deletion, insertion, substitution or addition of one or more nucleotides with respect to the nucleotide sequence represented by SEQ ID NO: 1, and encoding a polypeptide which has pyruvate decarboxylase activity;a polynucleotide encoding a polypeptide which consists of the amino acid sequence represented by SEQ ID NO: 2;a polynucleotide encoding a polypeptide which consists of an amino acid sequence having at least 80% identity to the amino acid sequence represented by SEQ ID NO: 2 and has pyruvate decarboxylase activity; anda polynucleotide encoding a polypeptide which consists of an amino acid sequence having deletion, insertion, substitution or addition of one or more amino acid residues with respect to the amino acid sequence represented by SEQ ID NO: 2 and has pyruvate decarboxylase activity.2Rhizopus. The mutant of the genus according to claim 1 , wherein the pyruvate decarboxylase activity is reduced to 50% or less as compared with that before mutation.3RhizopusRhizopusRhizopus oryzaeRhizopus delemar.. The mutant of the genus according to claim 1 , wherein the fungus of the genus is or4Rhizopus. The mutant of the genus according to claim 1 , wherein the mutant has improved productivity of an organic acid.5Rhizopus. The mutant of the genus according ...

ENGINEERING OF ACETYL-CoA METABOLISM IN YEAST

The invention relates to engineering of acetyl-CoA metabolism in yeast and in particular to production of acetyl-CoA in a non-ethanol producing yeast lacking endogenous gene(s) encoding pyruvate decarboxylase and comprising a heterologous pathway for synthesis of cytosolic acetyl-CoA. 122-. (canceled)23. A yeast comprising at least one heterologous pathway for synthesis of cytosolic acetyl-Coenzyme A (CoA) , the at least one heterologous pathway comprising at least one heterologous gene encoding a respective enzyme involved in synthesis of acetyl-CoA , wherein the at least one heterologous gene comprises:a heterologous gene encoding a pyruvate ferredoxin oxidoreductase;a heterologous gene encoding a ferredoxin reductase or flavodoxin reductase; and/ora heterologous gene encoding a ferredoxin reductase or encoding a flavodoxin reductase substrate, wherein the flavodoxin substrate is ferredoxin and/or flavodoxin.24. The yeast according to claim 23 , wherein{'i': 'Desulfovibrio africanus', 'said heterologous gene encoding said pyruvate ferredoxin oxidoreductase is a gene encoding a pyruvate ferredoxin oxidoreductase;'}{'i': 'Escherichia coli', 'said heterologous gene encoding said ferredoxin or flavodoxin reductase is a gene encoding an flavodoxin/ferredoxin reductase; and'}{'i': 'E. coli', 'said heterologous gene encoding said ferredoxin/flavodoxin reductase substrate is a heterologous gene encoding said flavodoxin/ferredoxin reductase substrate.'}25Desulfovibrio africanusD. africanus. The yeast according to claim 24 , wherein the pyruvate ferredoxin oxidoreductase is pfor;26Escherichia coliE. coli. The yeast according to claim 24 , wherein the flavodoxin/ferredoxin reductase is fpr.27E. coliE. coliE. coli. The yeast according to claim 24 , wherein gene encoding said flavodoxin/ferredoxin reductase substrate is fdx or fldA.28. The yeast according to claim 23 , wherein the yeast lacks any endogenous gene encoding pyruvate decarboxylase or comprises disrupted gene or ...

Genetically modified yeast species and fermenation processes using genetically modified yeast

Yeast cells are transformed with an exogenous xylose isomerase gene. Additional genetic modifications enhance the ability of the transformed cells to ferment xylose to ethanol or other desired fermentation products. Those modifications', include deletion of non-specific or specific aldose reductase gene(s), deletion of xylitol dehydrogenase gene(s) and/or overexpression of xylulokinase.

Yeast organism producing isobutanol at a high yield

There is disclosed a method of producing isobutanol. In an embodiment, the method includes providing a microorganism transformed with an isobutanol producing pathway containing at least one exogenous gene. The microorganism is selected to produce isobutanol from a carbon source at a yield of at least 10 percent theoretical. The method includes cultivating the microorganism in a culture medium containing a feedstock providing the carbon source, until isobutanol is produced. The method includes recovering the isobutanol. In one embodiment, the microorganism is a yeast with a Crabtree-negative phenotype. In another embodiment, the microorganism is a yeast microorganism with a Crabtree-positive phenotype. There is disclosed a microorganism for producing isobutanol. In an embodiment, the microorganism includes an isobutanol producing pathway containing at least one exogenous gene, and is selected to produce a recoverable quantity of isobutanol from a carbon source at a yield of at least 10 percent theoretical.

신규한 클루이베로마이세스 막시아누스 mj1 및 이의 용도

본 발명은 내열성 및 내산성을 나타내는 신규한 클루이베로마이세스 막시아누스( Kluyveromyces marxianus ) MJ1 균주, 상기 균주 및 이의 배양물을 포함하는 젖산 생산용 조성물 및 상기 균주를 배양하는 단계를 포함하는 젖산 생산 방법에 관한 것이다.

一种合成对羟基扁桃酸的方法

本发明公开了一种合成对羟基扁桃酸的方法，属于生物工程技术领域。本发明通过分子生物学手段构建了一株双质粒重组菌，共表达丙酮酸脱羧酶、苯乙醛脱氢酶和NADH氧化酶基因，将构建好的表达质粒导入E.coli BL21(DE3)。将对羟基苯甲醛和乙醛酸转化为对羟基扁桃酸，并通过NADH氧化酶辅酶再生系统，通过将NADH转化为NAD + ，使得NAD + 循环再生，转化能够高效进行。并通过优化底物浓度及比例，优化转化温度和pH，实现了对羟基扁桃酸的高效生产。在30℃、pH7的条件下反应24h，可将30mM的底物对羟基苯甲醛转化为28.5mM的对羟基扁桃酸，转化率可达到95％。

手性1，3-二羟基-1-芳基丙酮化合物的合成方法

本发明公布了生物催化剂催化羟基丙酮酸与不同取代基的苯甲醛反应生成光学纯手性1，3‑二羟基‑1‑取代芳基丙酮的方法。为了廉价的获得羟基丙酮酸，利用D‑氨基酸氧化酶转化D‑丝氨酸获得，不经分离，本方法具有反应条件温和、立体选择性好、无污染等显著特点。

一种产l-乳酸的重组菌及其应用

本发明公开了一种产L‑乳酸的重组菌及其应用，本发明的重组菌在酿酒酵母宿主中过表达了D‑乳酸脱氢酶Dld1基因，并采用来源于凝结芽孢杆菌的乳酸脱氢酶LDH基因替换丙酮酸脱羧酶Pdc1基因和丙酮酸脱羧酶Pdc6基因，采用来源于可合成L‑乳酸的乳酸乳球菌的L乳酸脱氢酶LLDH基因替换乙醇脱氢酶Adh1、甘油三磷酸脱氢酶1Gpd1或甘油三磷酸脱氢酶2Gpd2基因，敲除酿酒酵母中的转化乳酸为丙酮的L‑乳酸氧化还原酶基因Cyb2和羧基转运蛋白Jen1基因。通过本发明的生物改造技术，最终实现L‑LA的提升，摇瓶产量从最初的0.98g/L提升至29.55g/L，较原始耐酸菌株提高了30倍。

一种产d-1,2,4-丁三醇的基因工程菌及其应用

本发明公开了一种产D‑1,2,4‑丁三醇的基因工程菌及其应用,将2‑酮酸脱羧酶基因、木糖脱氢酶基因XylB、木糖酸脱水酶YjhG和醇脱氢酶基因YqhD转入宿主菌BL21（DE3）得到基因工程菌，所述2‑酮酸脱羧酶基因为来源于乳酸乳球菌（ Lactococcus lactis ）的α‑酮酸脱羧酶基因KdcA。经发酵培养生产D‑1,2,4‑丁三醇。本发明通过筛选所提供的α‑酮酸脱羧酶基因KdcA,能够有效提高D‑木糖合成D‑1,2,4‑丁三醇的能力，最终可达6.82g/L。

Methods and compositions for production of acetaldehyde

Microorganisms and methods for production of specific length fatty alcohols and related compounds

Genetically modified yeast species, and fermentation processes using genetically modified yeast

Yeast cells are transformed with an exogenous xylose isomerase gene. Additional genetic modifications enhance the ability of the transformed cells to ferment xylose to ethanol or other desired fermentation products. Those modifications include deletion of non-specific or specific aldose reductase gene(s), deletion of xylitol dehydrogenase gene(s) and/or overexpression of xylulokinase.

Genetically modified yeast species, and fermentation processes using genetically modified yeast

Yeast cells are transformed with an exogenous xylose isomerase gene. Additional genetic modifications enhance the ability of the transformed cells to ferment xylose to ethanol or other desired fermentation products. Those modifications include deletion of non-specific or specific aldose reductase gene(s), deletion of xylitol dehydrogenase gene(s) and/or overexpression of xylulokinase.