METABOLIC ENGINEERING FOR MICROBIAL PRODUCTION OF TERPENOID PRODUCTS

The invention relates to methods and bacterial strains for making terpene and terpenoid products, the bacterial strains having improved carbon pull through the MEP pathway and to a downstream recombinant synthesis pathway. 1. A method for production of a terpene or terpenoid product , comprising:providing a bacterial strain that produces isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) through an upstream methylerythritol phosphate pathway (MEP pathway) and converts the IPP and DMAPP to a terpene or terpenoid product through a downstream synthesis pathway;wherein IspG and IspH are overexpressed in the bacterial strain such that IspG activity and IspH activity are balanced to provide increased carbon flux to 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate (HMBPP) intermediate, while preventing accumulation of HMBPP at an amount that feeds back and reduces MEP pathway flux and terpene or terpenoid productivity, the strain optionally comprising one or more genetic modifications that enhance supply and/or transfer of electrons through the MEP pathway and/or to terpene and terpenoid products, and culturing the bacterial strain to produce the terpene or terpenoid product.2EscherichiaBacillusCorynebacteriumRhodobacterZymomonasVibrioPseudomonas. The method of claim 1 , wherein the bacterial strain is a bacteria selected from spp. claim 1 , spp. claim 1 , spp. claim 1 , spp. claim 1 , spp. claim 1 , spp. claim 1 , and spp.3Escherichia coli, Bacillus subtilis, Corynebacterium glutamicum, Rhodobacter capsulatus, Rhodobacter sphaeroides, Zvmomonas mobilis, Vibrio natriegensPseudomonas putida.. The method of claim 2 , wherein the bacterial strain is a species selected from claim 2 , or4E. coli.. The method of claim 3 , wherein the bacterial strain is5. The method of any one of to claim 3 , wherein the bacterial strain expresses dxs claim 3 , ispD claim 3 , ispF claim 3 , and idi as recombinant genes claim 3 , which are optionally expressed as an operon.6. The ...

Metabolic engineering for microbial production of terpenoid products

The invention relates to methods and bacterial strains for making terpene and terpenoid products, the bacterial strains having improved carbon pull through the MEP pathway and to a downstream recombinant synthesis pathway.

METHODS, CELLS AND REAGENTS FOR PRODUCTION OF ISOPRENE, DERIVATIVES AND INTERMEDIATES THEREOF

This application describes methods, including non-naturally occurring methods, for biosynthesizing 3-hydroxy-3-methylglutaryl-coA and intermediates thereof, as well as non-naturally occurring hosts for producing 3-hydroxy-3-methylglutaryl-coA. This application also describes methods, including non-naturally occurring methods, for biosynthesizing isoprene and intermediates thereof, as well as non-naturally occurring hosts for producing isoprene. 112-. (canceled)13. A non-naturally occurring host capable of producing 3-hydroxy-3-methylglutaryl-CoA , said host comprising:(a) at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.2.7.7 or EC 1.2.1.- enzyme; or at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.2.1.39 or EC 1.2.1.5 enzyme;', 'at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 6.2.1.2. enzyme; or (c) both (a) and (b); and, '(b) at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.1.1.74 or EC 4.1.1.43 enzyme;'}at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.3.8.4 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 6.4.1.4 enzyme; andat least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.2.1.18 enzyme.14. The host of claim 13 , wherein said host is capable of producing isoprene and comprises:at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.1.1.34 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 2.7.1.36 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 2.7.4.2 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.1.1.33 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 5.3.3.2 ...

RECOMBINANT STRAIN OF BACILLUS SUBTILIS

The invention relates to a recombinant strain of , wherein pyruvate carboxylase BalpycA, glyceraldehyde-3-phosphate ferredoxin dehydrogenase gor, isocitrate NAD dehydrogenase icd, malate quinone dehydrogenase mqo, pyruvate ferredoxin oxidoreductase porAB and nitrogenase ferritin cyh are integrated and expressed in the recombinant strain. The invention also discloses use of the recombinant strain in fermentation production of acetylglucosamine. The recombinant of the invention eliminates the central carbon metabolism overflow of the and balances the intracellular reducing force, and the fermentation yield of acetylglucosamine is greatly improved. 1Bacillus subtilis. A recombinant strain of , wherein pyruvate carboxylase BalpycA , glyceraldehyde-3-phosphate ferredoxin dehydrogenase gor , isocitrate NAD dehydrogenase icd , malate quinone dehydrogenase mqo , pyruvate ferredoxin oxidoreductase porAB and nitrogenase ferritin cyh are integrated and expressed in the recombinant strain.2Bacillus subtilis. The recombinant strain according to claim 1 , wherein the recombinant strain is obtained by using BSGNKAP2 as a starting strain.3. The recombinant strain according to claim 1 , wherein the pyruvate carboxylase BalpycA claim 1 , glyceraldehyde-3-phosphate ferredoxin dehydrogenase gor claim 1 , isocitrate NAD dehydrogenase icd claim 1 , malate quinone dehydrogenase mqo claim 1 , pyruvate ferredoxin oxidoreductase porAB and nitrogenase ferritin cyh are respectively expressed by using a strong constitutive promoter.4. The recombinant strain according to claim 3 , wherein the strong constitutive promoter is P claim 3 , P claim 3 , P claim 3 , P claim 3 , P claim 3 , P claim 3 , Por Ppromoter.5Bacillus subtilis. The recombinant strain according to any one of claim 1 , wherein the encoding gene balpycA of the pyruvate carboxylase BalpycA is shown as NCBI-Protein ID: AAS42897 claim 1 , and the encoding gene balpycA of the pyruvate carboxylase BalpycA is integrated into malS claim 1 ...

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 ...

Recombinant microorganisms exhibiting increased flux through a fermentation pathway

The invention provides methods of increasing the production of fermentation products by increasing flux through a fermentation pathway by optimising enzymatic reactions. In particular, the invention relates to identifying enzymes and/or co-factors involved in metabolic bottlenecks in fermentation pathways, and fermenting a CO-comprising substrate with a recombinant carboxydotrophic Clostridia microorganism adapted to exhibit increased activity of the one or more of said enzymes, or increased availability of the one or more of said co-factors, when compared to a parental microorganism. 1. A method of producing a fermentation product , the method comprising:a) determining a rate-limiting pathway reaction in a fermentation pathway;b) identifying at least one enzyme, co-factor or both, which are involved in catalysing the rate-limiting pathway reaction;c) fermenting a CO-comprising substrate with a recombinant carboxydotrophic Clostridia microorganism adapted to exhibit at least one of: i) increased activity of the at least one enzyme of (b) or a functionally equivalent variant thereof, or ii) increased availability of the at least one co-factor of (b), when compared to a parental microorganism, to produce a fermentation product.2. A method of producing a fermentation product , the method comprising fermenting a CO-comprising substrate with a recombinant carboxydotrophic Clostridia microorganism to produce a fermentation product , wherein the recombinant microorganism is adapted to exhibit at least one of:a) increased activity of at least one enzyme identified as being involved in catalysing a rate-limiting pathway reaction of a fermentation pathway, or a functionally equivalent variant thereof, when compared to a parental microorganism; orb) increased availability of at least one co-factor identified as being involved in catalysing a rate-limiting pathway reaction of a fermentation pathway, when compared to a parental microorganism.4. A recombinant carboxydotrophic ...

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.

GENETICALLY ENGINEERED BACTERIUM COMPRISING ENERGY-GENERATING FERMENTATION PATHWAY

The invention relates to a genetically engineered bacterium comprising an energy-generating fermentation pathway and methods related thereto. In particular, the invention provides a bacterium comprising a phosphate butyryltransferase (Ptb) and a butyrate kinase (Buk) (Ptb-Buk) that act on non-native substrates to produce a wide variety of products and intermediates. In certain embodiments, the invention relates to the introduction of Ptb-Buk into a C1-fixing microoorgansim capable of producing products from a gaseous substrate. 1. A genetically engineered bacterium comprising exogenous phosphate butyryltransferase (Ptb) and exogenous butyrate kinase (Buk) (Ptb-Buk) , wherein the Ptb-Buk acts on a non-native substrate to produce a non-native product.2. The bacterium of claim 1 , wherein the Ptb-Buk acts on a substrate other than butanoyl-CoA and/or butanoyl phosphate.3. The bacterium of claim 1 , wherein the Ptb-Buk produces a product other than butanoyl phosphate or butyrate.4. The bacterium of claim 1 , wherein the bacterium does not produce butanol.5. The bacterium of claim 1 , wherein the bacterium produces one or more of an acid claim 1 , an alkene claim 1 , a ketone claim 1 , an aldehyde claim 1 , an alcohol claim 1 , or a diol.6. The bacterium of claim 1 , wherein the bacterium produces one or more of acetone or a precursor thereof claim 1 , isopropanol or a precursor thereof claim 1 , isobutylene or a precursor thereof claim 1 , 3-hydroxybutyrate or a precursor thereof claim 1 , 1 claim 1 ,3-butanediol or a precursor thereof claim 1 , 2-hydroxyisobutyrate or a precursor thereof claim 1 , adipic acid or a precursor thereof claim 1 , 1 claim 1 ,3-hexanediol or a precursor thereof claim 1 , 3-methyl-2-butanol or a precursor thereof claim 1 , 2-buten-1-ol or a precursor thereof claim 1 , isovalerate or a precursor thereof claim 1 , or isoamyl alcohol or a precursor thereof.7. The bacterium of claim 1 , wherein the Ptb-Buk converts acetoacetyl-CoA to acetoacetate ...

GENETICALLY ENGINEERED BACTERIUM FOR THE PRODUCTION OF 3-HYDROXYBUTYRATE

The invention relates to a genetically engineered bacterium having an enzyme that converts acetyl-CoA to acetoacetyl-CoA, an enzyme that converts acetoacetyl-CoA to 3-hydroxybutyryl-CoA, and an enzyme that converts 3-hydroxybutyryl-CoA to 3-hydroxybutyrate. The bacterium may also have enzymes to produce other downstream products, such as 3-hydroxybutyryaldehyde, and 1,3-butanediol. Typically, the bacterium is capable of producing these products from a gaseous substrate, such as syngas or an industrial waste gas. 1. A genetically engineered C1-fixing bacterium comprising:(a) an enzyme that converts acetyl-CoA to acetoacetyl-CoA,(b) an enzyme that converts acetoacetyl-CoA to 3-hydroxybutyryl-CoA, and(c) an enzyme that converts 3-hydroxybutyryl-CoA to 3-hydroxybutyrate,wherein at least one of the enzymes is exogenous to the bacterium.2. The bacterium of claim 1 , wherein the enzyme that converts acetyl-CoA to acetoacetyl-CoA is thiolase (EC 2.3.1.9).3. The bacterium of claim 1 , wherein the enzyme that converts acetoacetyl-CoA to 3-hydroxybutyryl-CoA is 3-hydroxybutyryl-CoA dehydrogenase (EC 1.1.1.157) or acetoacetyl-CoA reductase (EC 4.2.1.36).4. The bacterium of claim 1 , wherein the enzyme that converts 3-hydroxybutyryl-CoA to 3-hydroxybutyrate is thioesterase (EC 3.1.2.20) claim 1 , phosphate butyryltransferase (EC 2.3.1.19) and butyrate kinase (EC 2.7.2.7) claim 1 , or CoA-transferase (EC 2.8.3.9).5. The bacterium of claim 1 , wherein the enzyme that converts 3-hydroxybutyryl-CoA to 3-hydroxybutyrate is stereospecific.6. The bacterium of claim 1 , wherein the 3-hydroxybutyrate is (R)-3-hydroxybutyrate claim 1 , (S)-3-hydroxybutyrate claim 1 , or a combination thereof.7. The bacterium of claim 1 , wherein the bacterium further comprises an isomerase that interconverts (R)-3-hydroxybutyrate and (S)-3-hydroxybutyrate.8. The bacterium of claim 1 , wherein the bacterium further comprises an enzyme that converts 3 -hydroxybutyrate to 3-hydroxybutyryaldehyde.9. The ...

MICROORGANISMS WITH ENHANCED CARBON MONOXIDE AVAILABILITY AND USE THEREOF

The present invention relates to a protein variant, a microorganism with enhanced carbon monoxide (CO) availability comprising the variant, and a use thereof. 1. A protein variant , wherein the 97residue from the N-terminus of SEQ ID NO: 1 , alanine (A) , is substituted with a different amino acid.2. The protein variant of claim 1 , wherein the different amino acid is selected from the group consisting of aspartic acid (D) claim 1 , glutamic acid (E) claim 1 , lysine (K) claim 1 , arginine (R) claim 1 , histidine (H) claim 1 , tyrosine (Y) claim 1 , asparagine (N) claim 1 , glutamine (Q) claim 1 , tryptophan (W) claim 1 , phenylalanine (F) claim 1 , methionine (M) claim 1 , and proline (P).3. The protein variant of claim 2 , wherein the different amino acid is glutamic acid (E).4. The protein variant of claim 1 , wherein the 290nucleotide in the nucleotide sequence of SEQ ID NO: 2 claim 1 , cytosine (C) claim 1 , is substituted with a different nucleotide.5. A microorganism comprising the protein variant of .6. The microorganism of claim 5 , wherein the microorganism is an acetogen.7Eubacterium limosum.. The microorganism of claim 6 , wherein the microorganism is8. The microorganism of claim 6 , wherein the microorganism is deposited under Accession No. KCTC 14201BP.9. A method for preparing a compound claim 5 , comprising a step of culturing the microorganism of .10. The method of claim 9 , wherein the microorganism is introduced with a gene encoding an enzyme involved in the synthesis of a compound.11. The method of claim 9 , wherein the compound is acetoin.12Bacillus subtilisAeromonas hydrophila,. The method of claim 10 , wherein the gene is an alsS gene derived from and an alsD gene derived from and the compound is acetoin.13. A method for preparing a microorganism with enhanced carbon monoxide (CO) availability claim 10 , which comprises a step of culturing the microorganism under a gas condition comprising carbon monoxide (CO).14. The method of claim 13 , ...

IMPROVED MICROBIAL PRODUCTION OF FATS

This invention describes a method of using microbial to produce fats, such as fatty acids and their derivatives, or products derived from the fatty acid synthesis cycle, such as hydroxyfatty acids, methyl ketones, and the like. 1. A method of producing products in bacteria , comprising:a) aerobically culturing a bacteria in a growth medium until sufficient cell mass is reached;b) smoothly transitioning from aerobic culturing to culturing under oxygen lean conditions (>0.1% DO to <5% DO) over a course of time, preferably 1-12 hrs, 2-8 hrs, or about 5 hrs;{'sub': '2', 'c) further culturing said bacteria under oxygen lean conditions by only sparging the head space with Ocontaining gas until product is formed; and'}d) isolating said product from said bacteria, said growth medium or both.2. The method of claim 1 , wherein in step a):{'sub': '600', 'a) aerobically culturing a bacteria in a growth medium with about 40% DO until an ODof >5 or >75%, preferably 80-90%, of maximal cell mass is reached;'}3. (canceled)4. The method of claim 1 , wherein in steps a)-c):{'sub': '600', 'a) aerobically culturing said bacteria in the growth medium with about 40% DO until an ODof >2 or >5 is reached;'}b) smoothly transitioning from about 40% DO to about 0.5% DO over a course of time, preferably 1-12 hrs, 2-8 hrs, or about 5 hrs;c) further culturing said bacteria with about 0.5% DO at about 350 rpm and only sparging the headspace with air until product is formed.5. The method of claim 1 , wherein in steps a) and c):a) aerobically culturing said bacteria in the growth medium until >75 to <95% maximum cell mass before stationary phase is reached;c) further culturing said bacteria with oxygen lean conditions and agitation at about 350 rpm and only sparging the head space with air until products are formed.6. (canceled)7. The method of claim 1 , wherein less pH control is used in said method than in a comparable method without said headspace sparging.8. (canceled)9. The method of claim 1 , ...

RECOMBINANT MICROORGANISMS EXHIBITING INCREASED FLUX THROUGH A FERMENTATION PATHWAY

The invention provides a recombinant, carboxydotrophic bacterium that expresses one or more of pyruvate:ferredoxin oxidoreductase (EC 1.2.7.1), acetolactate synthase (EC 2.2.1.6), and acetolactate decarboxylase (EC 4.1.1.5). The invention further provides a method of producing a fermentation product by fermenting the recombinant bacterium in the presence of a gaseous substrate comprising CO to produce one or more of ethanol, butanol, isopropanol, isobutanol, higher alcohols, butanediol, 2,3-butanediol, succinate, isoprenoids, fatty acids, biopolymers, and mixtures thereof. 1Clostridium. A recombinant , carboxydotrophic bacterium comprising one or more enzymes selected from the group consisting of pyruvate:ferredoxin oxidoreductase (EC 1.2.7.1) , acetolactate synthase (EC 2.2.1.6) , and acetolactate decarboxylase (EC 4.1.1.5) , wherein each enzyme is an overexpressed endogenous enzyme , a mutated endogenous enzyme , or an exogenous enzyme.2. The bacterium of claim 1 , comprising pyruvate:ferredoxin oxidoreductase and acetolactate synthase.3. The bacterium of claim 1 , comprising pyruvate:ferredoxin oxidoreductase and acetolactate decarboxylase.4. The bacterium of claim 1 , comprising acetolactate synthase and acetolactate decarboxylase.5. The bacterium of claim 1 , comprising pyruvate:ferredoxin oxidoreductase claim 1 , acetolactate synthase claim 1 , and acetolactate decarboxylase.6. The bacterium of claim 1 , wherein the enzyme is overexpressed endogenous pyruvate:ferredoxin oxidoreductase.7Desulfovibrio africanus. The bacterium of claim 1 , wherein the enzyme is exogenous pyruvate:ferredoxin oxidoreductase.8. The bacterium of claim 1 , wherein the enzyme is overexpressed endogenous IlvB ORF2059 acetolactate synthase claim 1 , overexpressed endogenous IlvB ORF2336 acetolactate synthase claim 1 , overexpressed endogenous IlvN acetolactate synthase claim 1 , or overexpressed endogenous AlsS acetolactate synthase.9. The bacterium of claim 1 , wherein the enzyme is ...

SYNTHETIC PATHWAY FOR BIOLOGICAL CARBON DIOXIDE SEQUESTRATION

This invention relates to methods for increasing carbon fixation and/or increasing biomass production in a plant, comprising: introducing into a plant, plant part, and/or plant cell heterologous polynucleotides encoding (1) a succinyl CoA synthetase, (2) a 2-oxoglutarate:ferredoxin oxidoreductase, (3) a 2-oxoglutarate carboxylase, (4) an oxalosuccinate reductase, or (5) an isocitrate lyase, or (6) a succinyl CoA synthetase and a 2-oxoglutarate:ferredoxin oxidoreductase, (7) a 2-oxoglutarate carboxylase and an oxalosuccinate reductase polypeptide, and/or (8) a 2-oxoglutarate carboxylase polypeptide, an oxalosuccinate reductase polypeptide and an isocitrate lyase polypeptide to produce a stably transformed plant, plant part, and/or plant cell, wherein said heterologous polynucleotides are from a bacterial and/or an archaeal species. Additionally, transformed plants, plant parts, and/or plant cells are provided as well as products produced from the transformed plants, plant parts, and/or plant cells. 1. A method for increasing carbon fixation and/or increasing biomass production in a plant , comprising:introducing into a plant, plant part, and/or plant cell a heterologous polynucleotide encoding a succinyl CoA synthetase polypeptide, a heterologous polynucleotide encoding a 2-oxoglutarate:ferredoxin oxidoreductase polypeptide, a heterologous polynucleotide encoding a 2-oxoglutarate carboxylase polypeptide, a heterologous polynucleotide encoding an oxalosuccinate reductase polypeptide, and/or a heterologous polynucleotide encoding an isocitrate lyase polypeptide to produce a stably transformed plant, plant part, and/or plant cell, wherein the heterologous polynucleotide encoding a succinyl CoA synthetase polypeptide, the heterologous polynucleotide encoding a 2-oxoglutarate:ferredoxin oxidoreductase polypeptide, the heterologous polynucleotide encoding a 2-oxoglutarate carboxylase polypeptide, the heterologous polynucleotide encoding an oxalosuccinate reductase ...

NOVEL CARBON FIXATION CYCLE AND USE THEREOF

The present invention relates to a novel carbon dioxide fixation cycle synthesizing a carbohydrate product from carbon dioxide in vitro. In addition, the present invention relates to a unit or a composition carrying out carbon dioxide fixation in cyclic manner. Additionally, the present invention relates to a method to fix carbon dioxide or a method to produce glyoxylate from the carbon dioxide fixation cycle. The present carbon dioxide fixation cycle is not found in natural world, and we found that, when the novel carbon dioxide fixation cycle is used, only three ATP molecules are consumed to fix one carbon dioxide molecule, and thus novel carbon dioxide fixation cycle has an energy conversion efficiency approximately 2.5 times higher than that of the Calvin cycle. 140-. (canceled)41. A novel carbon dioxide (CO) fixation cycle consisting of succinyl coenzyme A (succinyl-CoA) synthetase , 2-oxoglutarate synthase , isocitrate dehydrogenase , and isocitrate lyase , from which a carbohydrate product is formed from CO.42. The carbon dioxide fixation cycle of claim 41 , wherein succinyl-CoA synthetase converts succinate into succinyl-CoA claim 41 , 2-oxoglutarate synthase converts succinyl-CoA into 2-oxoglutarate claim 41 , isocitrate dehydrogenase converts 2-oxoglutarate into isocitrate claim 41 , and the isocitrate lyase converts isocitrate into succinate and glyoxylate.43. The carbon dioxide fixation cycle of claim 42 , wherein the concentrations of succinate or succinyl-CoA are maintained at higher level than that of 2-oxoglutarate or isocitrate.44. The carbon dioxide fixation cycle of claim 42 , wherein the concentrations of succinate and succinyl-CoA are maintained at ratio of 2:1 to 100:1.45. The carbon dioxide fixation cycle of claim 42 , wherein the concentrations of succinyl-CoA and 2-oxoglutarate are maintained at ratio of 100:1 to 10 claim 42 ,000:1.46. The carbon dioxide fixation cycle of claim 42 , wherein the concentrations of 2-oxoglutarate and isocitrate ...

METHODS AND COMPOSITIONS FOR THE AUGMENTATION OF PYRUVATE AND ACETYL-COA FORMATION

The present disclosure identifies methods and compositions for modifying photoautotrophic organisms as hosts, such that the organisms efficiently convert carbon dioxide and light into pyruvate or acetyl-CoA, and in particular the use of such organisms for the commercial production of molecules derived from these precursors, e.g., ethanol. 1. An engineered photosynthetic microbe , wherein said engineered photosynthetic microbe comprises a recombinant MdhP enzyme.2Pisum sativum. The engineered photosynthetic microbe of claim 1 , wherein said recombinant MdhP enzyme is a MdhP enzyme.3. The engineered photosynthetic microbe of claim 1 , wherein said recombinant MdhP enzyme is at least 95% identical to SEQ ID NO: 1.4. The engineered photosynthetic microbe of claim 1 , wherein said recombinant MdhP enzyme is at least 95% identical to SEQ ID NO: 2.5. The engineered photosynthetic microbe of claim 1 , wherein said engineered photosynthetic microbe comprises an additional mutation which reduces the expression or activity of its endogenous Mdh enzyme.6. The engineered photosynthetic microbe of claim 5 , wherein said mutation is a knockout of the gene encoding said endogenous Mdh enzyme.7. The engineered photosynthetic microbe of claim 1 , wherein said engineered photosynthetic microbe further comprises a recombinant phosphoenol pyruvate carboxylase.8. The engineered photosynthetic microbe of claim 1 , wherein said engineered photosynthetic microbe further comprises a recombinant NADPH-linked malic enzyme.9. The engineered photosynthetic microbe of claim 1 , wherein said engineered photosynthetic microbe further comprises a recombinant phosphoenol pyruvate carboxylase and a recombinant NADPH-linked malic enzyme.10. The engineered photosynthetic microbe of or claim 1 , wherein said recombinant phosphoenol pyruvate carboxylase is the S8D mutant phosphoenol pyruvate carboxylase.11Sorghum. The engineered photosynthetic microbe of claim 10 , wherein said S8D mutant phosphoenol ...

GENETICALLY ENGINEERED BACTERIUM FOR THE PRODUCTION OF ISOBUTYLENE

The invention relates to a genetically engineered bacterium having an enzyme that converts 3-hydroxyisovaleryl-CoA to 3-hydroxyisovalerate and an enzyme that converts 3-hydroxyisovalerate to isobutylene. Typically, the bacterium is capable of producing isobutylene from a gaseous substrate containing CO, CO, and/or H, such as syngas or an industrial waste gas. 2. The bacterium of claim 1 , wherein the enzyme that converts 3-hydroxyisovaleryl-CoA to 3-hydroxyisovalerate is thioesterase (EC 3.1.2.20) claim 1 , phosphate butyryltransferase (EC 2.3.1.19) and butyrate kinase (EC 2.7.2.7) claim 1 , or CoA-transferase (EC 2.8.3.9).3. The bacterium of claim 1 , wherein the enzyme that converts 3-hydroxyisovalerate to isobutylene is hydroxyisovalerate phosphorylase/decarboxylase or mevalonate diphosphate decarboxylase (EC 4.1.1.33).4. The bacterium of claim 1 , wherein the bacterium further comprises one or more enzymes selected from the group consisting of citramalate synthase (EC 2.3.1.182) claim 1 , 3-isopropylmalate dehydratase (EC 4.2.1.35) claim 1 , 3-isopropylmalate dehydrogenase (EC 1.1.1.85) claim 1 , acetolactate synthase (EC 2.2.1.6) claim 1 , ketol-acid reductoisomerase (EC 1.1.1.86) claim 1 , dihydroxyacid dehydratase (EC 4.2.1.9) claim 1 , ketoisovalerate oxidoreductase (EC 1.2.7.7) claim 1 , 2-methylbutanoyl-CoA dehydrogenase (EC 1.3.99.12) claim 1 , crotonase/3-hydroxybutyryl-CoA dehydratase (EC 4.2.1.55) claim 1 , crotonyl-CoA carboxylase-reductase (EC 1.3.1.86) claim 1 , crotonyl-CoA reductase (EC 1.3.1.44) claim 1 , 3-hydroxypropionyl-CoA dehydratase (EC 4.2.1.116) claim 1 , enoyl-CoA hydratase (4.2.1.17) claim 1 , thiolase (EC 2.3.1.9) claim 1 , thioesterase (EC 3.1.2.20) claim 1 , phosphate butyryltransferase (EC 2.3.1.19) claim 1 , butyrate kinase (EC 2.7.2.7) claim 1 , or CoA-transferase (EC 2.8.3.9) claim 1 , acetoacetate decarboxylase (EC 4.1.1.4) claim 1 , alpha-ketoisovalerate decarboxylase (EC 4.1.1.74) claim 1 , 3-hydroxyisovaleryl-CoA synthase ...

METABOLIC ENGINEERING FOR MICROBIAL PRODUCTION OF TERPENOID PRODUCTS

The invention relates to methods and bacterial strains for making terpene and terpenoid products, the bacterial strains having improved carbon pull through the MEP pathway and to a downstream recombinant synthesis pathway. 1. A method for production of a terpene or terpenoid product , comprising:providing a bacterial strain that produces isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) through an upstream methylerythritol phosphate pathway (MEP pathway) and converts the IPP and DMAPP to a terpene or terpenoid product through a downstream synthesis pathway;wherein IspG and IspH are overexpressed in the bacterial strain such that IspG activity and IspH activity are balanced to provide increased carbon flux to 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate (HMBPP) intermediate, while preventing accumulation of HMBPP at an amount that feeds back and reduces MEP pathway flux and terpene or terpenoid productivity; the strain optionally comprising one or more genetic modifications that enhance supply and/or transfer of electrons through the MEP pathway and/or to terpene and terpenoid products, and culturing the bacterial strain to produce the terpene or terpenoid product.2EscherichiaBacillusCorynebacteriumRhodobacterZymomonasVibrioPseudomonas. The method of claim 1 , wherein the bacterial strain is a bacteria selected from spp. claim 1 , spp. claim 1 , spp. claim 1 , spp. claim 1 , spp. claim 1 , spp. claim 1 , and spp.3. (canceled)4E. coli.. The method of claim 2 , wherein the bacterial strain is5. The method of claim 1 , wherein the bacterial strain expresses dxs claim 1 , ispD claim 1 , ispF claim 1 , and idi as recombinant genes claim 1 , which are optionally expressed as an operon.6. The method of claim 5 , wherein the bacterial strain expresses dxs claim 5 , dxr claim 5 , ispD claim 5 , ispE claim 5 , ispF claim 5 , and idi as recombinant genes claim 5 , which are optionally expressed as 1 claim 5 , 2 claim 5 , or 3 operons.7. (canceled)8. The ...

MICROBIAL ORGANISMS FOR CONVERTING ACETYL-COA INTO CROTYL ALCOHOL AND METHODS FOR PRODUCING CROTYL ALCOHOL

The present invention provides microorganisms capable of converting acetyl-coA into crotyl alcohol as well as fermentation methods for producing crotyl alcohol, either alone, or in combination with acetone and/or isopropanol. The microorganisms may be genetically engineered to express and/or disrupt one or more of the following enzymes: acetaldehyde dehydrogenase, alcohol dehydrogenase, bifunctional acetaldehyde/alcohol dehydrogenase, aldehyde oxidoreductase, phosphotransacetylase, acetate kinase, CoA-transferase A, CoA-transferase B, acetoacetate decarboxylase, secondary alcohol dehydrogenase, butyryl-CoA dehydro genase (BCD), and/or trans-2-enoyl-CoA reductase (TER). 1. A non-naturally occurring microbial organism capable of converting acetyl-CoA into crotyl alcohol , wherein at least one of the following genes are deleted , disrupted or silenced and/or expression from at least one of the following genes is disrupted or silenced:i. Butyryl-CoA dehydrogenase (BCD); and/orii. Trans-2-enoyl-CoA reductase (TER).2 claim 1 , A microbial organism according to claim 1 , comprising a disrupted claim 1 , deleted claim 1 , or mutated BCD and/or TER gene.3. A microbial organism according to claim 1 , wherein disruption or silencing of expression includes disruption or silencing of RNA transcription and/or protein translation.4. A microbial organism according to claim 1 , wherein disruption or silencing of expression comprises protein translation silencing using RNA interference.5. A microbial organism according to claim 1 , comprising at least one exogenous nucleic acid encoding one or more of the following enzymes fbr producing crotyl alcohol from crotonyl-CoA:A. Acetaldehyde dehydrogenase;B. Alcohol dehydrogenase;C, Bifunctional acetaldehyde/alcohol dehydrogenase;D. Aldehyde oxidoreductase;E. Pbosphotransacetylase; and/orF. Acetate kinase.6. A microbial organism according to for further producing acetone and/or isopronanol claim 5 , comprising at least a second exogenous ...

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 ...

Method for Producing Aldehyde

A method is described for producing an objective substance, for example, an aldehyde such as vanillin. The objective substance is produced from a carbon source or a precursor of the objective substance by using a microorganism having an ability to produce the objective substance, wherein the microorganism has been modified to have a specific carboxylic acid reductase (CAR) gene, such as a CAR gene, CAR gene, or CAR gene. 1. A method for producing an objective substance , the method comprising:producing the objective substance by using a microorganism having an ability to produce the objective substance,wherein the objective substance is an aldehyde,wherein the microorganism has been modified to have an aldehyde oxidoreductase gene, andwherein the aldehyde oxidoreductase gene encodes a protein selected from the group consisting of:(a) a protein comprising the amino acid sequence of SEQ ID NO: 18, 79, or 83;(b) a protein comprising the amino acid sequence of SEQ ID NO: 18, 79, or 83 but which includes substitution, deletion, insertion, and/or addition of 1 to 10 amino acid residues, and wherein said protein has aldehyde oxidoreductase activity;(c) a protein comprising an amino acid sequence having an identity of 90% or higher to the amino acid sequence of SEQ ID NO: 18, 79, or 83, and wherein said protein has aldehyde oxidoreductase activity.2. The method according to claim 1 , wherein said producing comprises:cultivating the microorganism in a culture medium containing a carbon source to produce and accumulate the objective substance in the culture medium.3. The method according to claim 1 , wherein said producing comprises:converting a precursor of the objective substance into the objective substance by using the microorganism.4. The method according to claim 3 , wherein said converting comprises:cultivating the microorganism in a culture medium containing the precursor to produce and accumulate the objective substance in the culture medium.5. The method according ...

Genetically engineered microbes and methods for converting organic acids to alcohols

Disclosed herein are genetically engineered microbes. In one embodiment, a genetically engineered microbe includes a metabolic pathway for the production of an alcohol from an organic acid. For instance, a genetically engineered microbe converts acetate, butyrate, propionate, isobutyrate, valerate, isovalerate, caproate, or phenylacetate, to Ethanol, Butanol, Propanol, Isobutanol, 1-Pentanol, Isoamylalcohol, 1-Hexanol, Phenylethanol, respectively. Also provided herein are methods of using the microbes.

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 ...

RECOMBINANT CELL, AND METHOD FOR PRODUCING ISOPRENE

To provide a series of techniques capable of producing isoprene from syngas or the like. 120-. (canceled)21. A recombinant cell prepared by introducing a nucleic acid encoding isoprene synthase into a host cell having an isopentenyl diphosphate synthesis ability by a non-mevalonate pathway , wherein the nucleic acid is expressed in the host cell , and the recombinant cell is capable of producing isoprene from at least one C1 compound selected from the group consisting of carbon monoxide , carbon dioxide , formic acid , and methanol.22. The recombinant cell according to claim 21 , having carbon monoxide dehydrogenase.23ClostridiumMoorella. The recombinant cell according to claim 21 , wherein the host cell is a bacterium or a bacterium.24. The recombinant cell according to claim 21 , wherein a nucleic acid encoding a group of enzymes acting in a mevalonate pathway is further introduced so that an isopentenyl diphosphate synthesis ability by a mevalonate pathway is further imparted.25. The recombinant cell according to claim 24 , wherein the mevalonate pathway is that of yeast claim 24 , prokaryote or actinomycete.26. The recombinant cell according to claim 21 , wherein a nucleic acid encoding at least one enzyme acting in a non-mevalonate pathway is further introduced claim 21 , and the nucleic acid is expressed in the host cell.27. The recombinant cell according to claim 26 , wherein the non-mevalonate pathway is that of other organism than the host cell.28. The recombinant cell according to claim 21 , wherein the isoprene synthase is derived from plant.29. The recombinant cell according to claim 21 , wherein the nucleic acid encoding isoprene synthase encodes the following (a) claim 21 , (b) or (c):(a) a protein consisting of an amino acid sequence represented by SEQ ID NO: 2,(b) a protein consisting of an amino acid sequence in which 1 to 20 amino acids are deleted, substituted or added in the amino acid sequence represented by SEQ ID NO: 2, and having isoprene ...

METHODS, CELLS AND REAGENTS FOR PRODUCTION OF ISOPRENE, DERIVATIVES AND INTERMEDIATES THEREOF

This application describes methods, including non-naturally occurring methods, for biosynthesizing 3-hydroxy-3-methylglutaryl-coA and intermediates thereof, as well as non-naturally occurring hosts for producing 3-hydroxy-3-methylglutaryl-coA. This application also describes methods, including non-naturally occurring methods, for biosynthesizing isoprene and intermediates thereof, as well as non-naturally occurring hosts for producing isoprene. 110.-. (canceled)11. A non-naturally occurring host capable of producing 3-hydroxy-3-methylglutaryl-CoA , said host comprising:at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.3.8.4 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 6.4.1.4 enzyme; andat least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.2.1.18 enzyme;and wherein said host further comprises:(a) at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.2.7.7 or EC 1.2.1.-enzyme; at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.2.1.39 or EC 1.2.1.5 enzyme, and', 'at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 6.2.1.2. enzyme; or, '(b) at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.1.1.74 or EC 41.2.1.43 enzyme,'} at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.1.1.74 or EC 41.2.1.43 enzyme,', 'at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.2.1.39 or EC 1.2.1.5 enzyme, and', 'at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 6.2.1.2. enzyme., '(c) at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.2.7.7 or EC 1.2.1.-enzyme,'}1213.-. (canceled)14. The host of claim 11 , wherein said host is capable of producing isoprene and ...

Compositions and Methods for the Conversion of Short-Chained Carboxylic Acids to Alcohols Using Clostridial Enzymes

The invention relates to the fields of bacterial metabolism and the utilization or consumption of short-chain carboxylic acids to reduced products. Specifically, it relates to syngas fermentations using monocultures of syngas-utilizing homoacetogenic bacteria for the production of alcohols using native alcohol dehydrogenase.

Method for Promoting Acetylglucosamine Synthesis of Bacillus Subtilis

The present invention relates to a method for promoting acetylglucosamine synthesis of , which belongs to the field of genetic engineering. The present invention adopts the recombinant BSGNKAP2 as a starting strain, exogenously introducing pyruvate carboxylase BalpycA derived from , eliminating the central carbon metabolism overflow of the and avoiding the synthesis of the by-product acetoin; further, five exogenous reducing force metabolic reactions are introduced to replace the reaction of generating NADH in glycolysis pathway and tricarboxylic acid cycle to reconstruct intracellular reducing force metabolism, which specifically comprise glyceraldehyde-3-phosphate ferredoxin dehydrogenase, isocitrate NAD dehydrogenase, a malate quinone dehydrogenase, a ketoacid ferredoxin oxidoreductase and a nitrogenase ferritin. In a shake-flask fermentation process using a complex medium, acetylglucosamine yield of the recombinant strain BSGNKAP8 is 24.50 g/L, acetylglucosamine/glucose yield is 0.469 g/g, respectively 1.97 times and 2.13 times of those of the starting strain BSGNKAP2. 1Bacillus subtilis. A recombinant strain of , wherein the recombinant strain comprises an integrated expression sequence capable of expressing pyruvate carboxylase BalpycA , glyceraldehyde-3-phosphate ferredoxin dehydrogenase gor , isocitrate NAD dehydrogenase icd , malate quinone dehydrogenase mqo , pyruvate ferredoxin oxidoreductase porAB and nitrogenase ferritin cyh.2Bacillus subtilis. The recombinant strain according to claim 1 , wherein the recombinant strain is configured to adopt BSGNKAP2 as a starting strain.3Bacillus cereus. The recombinant strain according to claim 1 , wherein the pyruvate carboxylase BalpycA is derived from claim 1 , and the pyruvate carboxylase BalpycA is set forth as NCBI-Protein ID: AAS42897.1.4. The recombinant strain according to claim 1 , wherein the pyruvate carboxylase BalpycA encoding gene balpycA is configured to be expressed by using a strong constitutive ...

RECOMBINANT CELLS AND METHOD FOR PRODUCING ISOPRENE OR TERPENE

To provide a recombinant cell being an anaerobic archaeon, including a gene encoding isoprene synthase, a gene encoding monoterpene synthase, a gene encoding sesquiterpene synthase, a gene encoding diterpene synthase, a gene encoding squalene synthase, or a gene encoding phytoene synthase as a first foreign gene, wherein the first foreign gene is expressed, and the recombinant cell is capable of producing isoprene or terpene having 10, 15, 20, 30, or 40 carbon atoms. 1. A recombinant cell being an anaerobic archaeon , comprising: a gene encoding isoprene synthase , a gene encoding monoterpene synthase , a gene encoding sesquiterpene synthase , a gene encoding diterpene synthase , a gene encoding squalene synthase , or a gene encoding phytoene synthase as a first foreign gene ,wherein the first foreign gene is expressed, and the recombinant cell produces isoprene or terpene having 10, 15, 20, 30, or 40 carbon atoms.2. The recombinant cell according to claim 1 , growing using at least one selected from the group consisting of carbon monoxide claim 1 , carbon dioxide claim 1 , methane claim 1 , methanol claim 1 , and acetic acid as a sole carbon source.3. The recombinant cell according to claim 1 , comprising carbon monoxide dehydrogenase.4. The recombinant cell according to claim 1 , having a function of synthesizing acetyl-CoA from methyltetrahydropterin claim 1 , carbon monoxide claim 1 , and CoA.5. The recombinant cell according to claim 1 , having a methane formation potential.6. The recombinant cell according to claim 1 , growing using carbon dioxide as a sole carbon source and hydrogen as an energy source.7. The recombinant cell according to claim 1 , growing using methanol as a sole carbon source.8. (canceled)9MethanosarcinaMethanococcusMethanothermococcusMethanothermobacterMethanothrixThermococcusThermofilumArchaeoglobus.. The recombinant cell according to claim 1 , belonging to the genus claim 1 , the genus claim 1 , the genus claim 1 , the genus claim 1 , ...

MICROORGANISM WITH MODIFIED ALDEHYDE:FERREDOXIN OXIDOREDUCTASE ACTIVITY AND RELATED METHODS

The invention provides a non-naturally occurring bacterium having decreased or eliminated activity of an enzyme that catalyzes the reaction defined by EC 1.2.7.5, such as aldehyde:ferredoxin oxidoreductase (AOR). Optionally, the bacterium also has decreased or eliminated activity of an enzyme that catalyzes the reaction defined by EC 1.2.1.10 and/or EC 1.1.1.1, such as aldehyde dehydrogenase, alcohol dehydrogenase, or bifunctional aldehyde/alcohol dehydrogenase. The invention further provides methods of producing products by culturing the bacterium in the presence of a gaseous substrate containing one or more of CO, CO, and H. 1. A non-naturally occurring bacterium having decreased or eliminated activity of an enzyme that catalyzes the reaction defined by EC 1.2.7.5 compared to a parental bacterium.2. The non-naturally occurring bacterium of claim 1 , wherein the non-naturally occurring bacterium comprises at least one disruptive mutation in a gene encoding the enzyme that catalyzes the reaction defined by EC 1.2.7.5.3. The non-naturally occurring bacterium of claim 1 , wherein the enzyme that catalyzes the reaction defined by EC 1.2.7.5 is aldehyde:ferredoxin oxidoreductase.4. The non-naturally occurring bacterium of claim 1 , wherein the non-naturally occurring bacterium further has decreased or eliminated activity of at least one enzyme that catalyzes the reaction defined by EC 1.2.1.10 and/or EC 1.1.1.1 compared to the parental bacterium.5. The non-naturally occurring bacterium of claim 4 , wherein the non-naturally occurring bacterium comprises at least one disruptive mutation in a gene encoding the enzyme that catalyzes the reaction defined by EC 1.2.1.10 and/or EC 1.1.1.1.6. The non-naturally occurring bacterium of claim 4 , wherein the enzyme that catalyzes the reaction defined by EC 1.2.1.10 and/or EC 1.1.1.1 is selected from the group consisting of bifunctional aldehyde/alcohol dehydrogenase claim 4 , aldehyde dehydrogenase claim 4 , and alcohol ...

Method for increasing growth and metabolism efficiency of recombinant microorganism under anaerobic environment

The present invention provides a method for increasing the metabolic rate of recombinant microorganism growth under an anaerobic environment, wherein a recombinant strain is placed under an anaerobic environment and cultured under a culture condition, wherein the culture condition includes a potential difference and a nitrogen source, but not includes an organic carbon source. According to the method disclosed by the present invention, the recombinant strain can perform anaerobic respiration and metabolic reaction in an anaerobic environment, and can grow stably and rapidly.

Biological Conversion and Product Recovery Processes

The invention provides a process for reducing bio-catalytic oxidation of a product in a post-production stream. More particularly the invention provides a process for reducing bio-catalytic oxidation of an alcohol in a product stream, the product stream comprising an alcohol product, dissolved carbon dioxide, and at least one enzyme capable of oxidizing the alcohol. The invention finds applicability in fermentation processes, wherein a C1-fixing microorganism utilizes a C1-containing substrate to produce a fermentation product.

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 ...

呈现通过发酵路径的增大的通量的重组微生物

本发明提供一种重组、一氧化碳营养型梭菌属细菌，其表达丙酮酸:铁氧还蛋白氧化还原酶(EC 1.2.7.1)、乙酰乳酸合酶(EC 2.2.1.6)和乙酰乳酸脱羧酶(EC 4.1.1.5)中的一种或多种。本发明进一步提供一种生产发酵产物的方法，其通过在包含CO的气态底物存在下发酵重组细菌以生产乙醇、丁醇、异丙醇、异丁醇、高级醇、丁二醇、2,3‑丁二醇、丁二酸酯、类异戊二烯、脂肪酸、生物聚合物及其混合物中的一种或多种。

Recombinant microorganisms exhibiting increased flux through a fermentation pathway

The invention provides methods of increasing the production of fermentation products by increasing flux through a fermentation pathway by optimising enzymatic reactions. In particular, the invention relates to identifying enzymes and/or co-factors involved in metabolic bottlenecks in fermentation pathways, and fermenting a CO-comprising substrate with a recombinant carboxydotrophic Clostridia microorganism adapted to exhibit increased activity of the one or more of said enzymes, or increased availability of the one or more of said co-factors, when compared to a parental microorganism.

Genetically engineered bacterium with altered carbon monoxide dehydrogenase (CODH) activity

The invention provides genetically engineered microorganisms with altered carbon monoxide dehydrogenase (CODH) activity and methods related thereto. In particular, the invention provides a genetically engineered carboxydotrophic acetogenic bacterium having decreased or eliminated activity of CODH1 and/or CODH2. In certain embodiments, the bacterium may also have increased activity of CODH/ACS. The invention further provides a method for producing a product by culturing the bacterium in the presence of a gaseous substrate comprising one or more of carbon monoxide, carbon dioxide, and hydrogen.

Compositions and methods for the conversion of short-chained carboxylic acids to alcohols using clostridial enzymes

The invention relates to the fields of bacterial metabolism and the utilization or consumption of short-chain carboxylic acids to reduced products. Specifically, it relates to syngas fermentations using monocultures of syngas-utilizing homoacetogenic bacteria for the production of alcohols using native alcohol dehydrogenase.

Genetically engineered bacterium comprising energy-generating fermentation pathway

The invention relates to a genetically engineered bacterium comprising an energy-generating fermentation pathway and methods related thereto. In particular, the invention provides a bacterium comprising a phosphate butyryltransferase (Ptb) and a butyrate kinase (Buk) (Ptb-Buk) that act 011 non-native substrates to produce a wide variety of products and intermediates. In certain embodiments, the invention relates to the introduction of Ptb-Buk into a C1-fixing microoorgansim capable of producing products from a gaseous substrate.

Microorganism with modified aldehyde:ferredoxin oxidoreductase activity and related methods

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.

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.

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.

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.

Recombinant microorganisms exhibiting increased flux through a fermentation pathway

The invention provides a recombinant, carboxydotrophic Clostridium bacterium that expresses one or more of pyruvate:ferredoxin oxidoreductase (EC 1.2.7.1), acetolactate synthase (EC 2.2.1.6), and acetolactate decarboxylase (EC 4.1.1.5). The invention further provides a method of producing a fermentation product by fermenting the recombinant bacterium in the presence of a gaseous substrate comprising CO to produce one or more of ethanol, butanol, isopropanol, isobutanol, higher alcohols, butanediol, 2,3-butanediol, succinate, isoprenoids, fatty acids, biopolymers, and mixtures thereof.

Methods and Compositions for Improving the Production of Fuels in Microorganisms

The invention relates to compositions, systems, and methods for producing fuels, such as ethanol and hydrogen, and related compounds. More specifically, compositions and methods are provided for making recombinant microorganisms for the production of fuels using genes from the Clostridium phytofermentans ethanol and hydrogen pathways disclosed herein.

Synthetic pathways for biofuel synthesis

Enhanced Butanol Producing Recombinant Microorganisms and Method for Preparing Butanol Using the Same

본 발명은 부탄올 생성능이 개선된 재조합 미생물 및 이를 이용한 부탄올의 제조방법에 관한 것으로서, 더욱 상세하게는 L-발린의 전구체인 2-케토이소발레르산염을 중간체(intermediate)로 하여 부탄올을 생합성하는 재조합 미생물 및 이를 이용한 부탄올의 제조방법에 관한 것이다. 본 발명에 따른 재조합 미생물은 L-발린의 전구체인 2-케토이소발레르산염의 생성능을 가지는 미생물에, 2-케토이소발레르산염을 이소부티릴-CoA로 전환시키는 효소인 VorABC 효소를 코딩하는 유전자, 이소부티릴-CoA를 부티릴-CoA로 전환시키는 효소를 코딩하는 유전자, 부티릴-CoA를 부틸알데하이드로 전환시키는 효소를 코딩하는 유전자 및 부틸알데하이드를 부탄올로 전환시키는 효소를 코딩하는 유전자가 도입되어 있는 것을 특징으로 한다. 본 발명에 따른 재조합 미생물은 기존의 클로스트리디움 아세토부틸리쿰( Clostridium acetobutylicum) 과 달리 성장이 용이하고, 추가 대사흐름 조작으로 인한 균주개량의 가능성이 크므로 부탄올의 산업적 생성 미생물로 추가 대사흐름 조작으로 인한 균주개량의 가능성이 크므로 부탄올의 산업적 생성 미생물로 유용하며, 2-케토이소발레르산염을 이소부티릴-CoA로 전환시키는 과정을 단순화시킴으로써 부탄올 생성능이 개선된 재조합 미생물을 제공하는 효과가 있다. The present invention relates to a recombinant microorganism having improved butanol production ability, and a method for producing butanol using the recombinant microorganism. More particularly, the present invention relates to a recombinant microorganism having improved butanol production ability by using 2-ketoisovalerate as a precursor of L-valine as an intermediate, And a method for producing butanol using the microorganism. The recombinant microorganism according to the present invention is characterized in that a microorganism having the ability to produce 2-ketoisovalerate as a precursor of L-valine is added to a microorganism having the ability to produce 2-ketoisovalerate as a microorganism which encodes a VorABC enzyme which is an enzyme for converting 2-ketoisovalerate into isobutyryl- A gene encoding an enzyme that converts isobutyryl-CoA into butyryl-CoA, a gene that codes an enzyme that converts butyryl-CoA into butylaldehyde, and a gene that encodes an enzyme that converts butylaldehyde to butanol Is introduced. Since the recombinant microorganism according to the present invention is easy to grow unlike the conventional Clostridium acetobutylicum and has a high possibility of improving the strain due to the manipulation of the additional metabolic flow, the recombinant microorganism according to ...

Transgenic Eubacterium genus strain with improved acetic acid productivity and cell growth, method for manufacturing the same

One embodiment of the present invention provides a transgenic eubacterium genus strain with improved acetic acid productivity and cell growth compared with the strain before mutation through the joint overexpression of carbon monoxide dehydrogenase and coenzyme thereof, and a manufacturing method thereof. Using the transgenic eubacterium genus strain according to an embodiment of the present invention, it is possible to convert C1 gas in a low-cost process with improved reaction rate and reaction efficiency.

Method for mass producing human blood coagulation factor vii derivative

本文提供了制备下列化合物的方法、组合物和非天然存在的微生物有机体，所述化合物例如为1丁醇、丁酸、琥珀酸、1,4丁二醇、1戊醇、戊酸、戊二酸、1,5戊二醇、1己醇、己酸、己二酸、1,6己二醇、6羟基己酸、ε己内酯、6氨基己酸、ε己内酰胺、六亚甲基二胺、7～25个碳原子长之间的直链脂肪酸和直链脂肪醇、6～24个碳原子长之间的直链烷烃和直链α烯烃、癸二酸和十二烷二酸，所述方法包括：a)通过羟醛加成将C N 醛和丙酮酸转化为C N+3 β羟基酮中间体；以及b)通过酶促步骤、或酶促步骤和化学步骤的组合，将C N+3 β羟基酮中间体转化为所述化合物。

Recombinant host cells comprising phosphoketolases

本发明涉及重组宿主细胞，其包含：（i）编码多肽的内源性基因中的至少一个缺失、突变或替换，所述多肽将丙酮酸转化为乙醛、乙酰-磷酸或乙酰-CoA；和（ii）编码具有磷酸酮醇酶活性的多肽的异源性多核苷酸。本发明也涉及重组宿主细胞，其还包含（iii）编码具有磷酸转乙酰酶活性的多肽的异源性多核苷酸。

Light Inducible Promoter Enhanced Light Inducible Activity and Gene Expression System Comprising The Same

본 발명은 SORLIP 엘리먼트를 이용한 개량 프로모터 및 이를 포함하는 유전자 발현 시스템에 관한 것이다. 본 발명의 광유도 활성 증진용 폴리뉴클레오티드를 이용하는 경우, SORLIP 엘리먼트를 포함하는 광유도성 프로모터의 광유도 활성을 크게 증진시킬 수 있고, 본 발명의 형질전환체를 이용하는 경우, 목적 단백질의 발현량을 광 조사에 의해 조절하면서 목적 단백질을 효과적으로 발현 시킬 수 있다.

Methods and compositions for improving the production of fuels in microorganisms

The invention relates to compositions, systems, and methods for producing fuels, such as ethanol and hydrogen, and related compounds. More specifically, compositions and methods are provided for making recombinant microorganisms for the production of fuels using genes from the Clostridium phytofermentans ethanol and hydrogen pathways disclosed herein.

An artificial CO2 fixation cycle and its uses

The present invention relates to a novel carbon dioxide fixing circuit for fixing carbohydrates by fixing carbon dioxide. The present invention also relates to a carbon dioxide fixing performance unit or composition comprising the carbon dioxide fixing circuit. In addition, the present invention relates to a method for fixing carbon dioxide using the carbon dioxide fixing circuit or a method for producing glyoxylic acid. The use of the new carbon dioxide fixing circuit according to the present invention consumes only three ATPs for fixing one carbon dioxide, and thus energy conversion efficiency about 2.5 times higher than that of a typical carbon dioxide-fixing enzyme, Rubisco enzyme, can be realized. In addition, when the above-described new carbon dioxide fixing circuit is operated outside the living body, the chemical energy generated from the light reaction of photosynthesis can be entirely used only for carbon compound synthesis without energy loss occurring in cell maintenance and other metabolic processes, . In addition, the novel carbon dioxide fixing circuit according to the present invention can produce glyoxylic acid by continuously fixing carbon dioxide in addition to the carbohydrate that is initially added, without providing any additional carbohydrate.

Pasteurella GAPDH molecular antigen polypeptide for serum antibody detection and preparation method and application thereof

本发明属于生物医学技术领域，提供了一种用于血清抗体检测的巴氏杆菌GAPDH分子抗原多肽，所述GAPDH分子抗原多肽包含巴氏杆菌GAPDH表位，其氨基酸序列为如下三段序列之一，PMG‑2A：HATTATQKTVDGPSAKDWRGGRGA；PMG‑2B：TQKTVDGPSAKDWRGGRGAAQNIIP；PMG‑2D：KDWRGGRGAAQNIIPSSTGA。上述巴氏杆菌的GAPDH分子抗原表位多肽与养殖场常见细菌高免血清均不发生反应，具有高度的特异性，可以用于建立评估巴氏杆菌GAPDH免疫后血清抗体水平的检测方法。

Fermentation preparation of 2-phenylethanol from gaseous substrates

본원에 개시된 것은 일산화탄소 및/또는 이산화탄소를 포함하는 기질의 미생물 발효에 의한 2-페닐에탄올의 제조 방법이고, 추가로 개시된 것은 상기 방법에서 사용하기 위한 유전자 변형된 미생물이다. 부가적으로, 본원에 개시된 방법은 천연 및 석유화학 공정에 대한 의존성을 경감시키는 개선된 2-PE 제조 방법이다.

Corynebacterium glutamicum capable of producing L-proline in high yield and method for producing L-proline in high yield

本发明提供了一种生产L‑脯氨酸的谷氨酸棒杆菌，以及利用该菌株生产L‑脯氨酸的方法。本发明构建的生产L‑脯氨酸的谷氨酸棒杆菌，其中脯氨酸脱氢酶/吡咯‑5‑羧酸脱氢酶PutA失活，谷氨酸激酶ProB、谷氨酸‑5‑半醛脱氢酶ProA、吡咯‑5‑羧酸脱氢酶ProC、丙酮酸羧化酶Pyc、甘油醛‑3‑磷酸脱氢酶GapN、L‑脯氨酸外排蛋白ThrE或SerE活性增强，L‑谷氨酸外排蛋白MscCG失活，得到的菌株的L‑脯氨酸产量、转化率和生产强度较出发菌株显著提升，可降低L‑脯氨酸的生产成本。

high-yield route for the production of compounds from renewable sources

“rota de alto rendimento para a produção de compostos a partir de fontes renováveis” resumo são fornecidos aqui métodos, composições e organismos microbianos que não ocorrem naturalmente, para preparar compostos tais como 1-butanol, ácido butírico, ácido succínico, 1,4-butanodiol, 1-pentanol, ácido pentanóico, ácido glutárico, 1,5-pentanodol, 1-hexanol, ácido exanóico, ácido adípico, 1,6-hexanodiol, ácido 6-hidroxi hexanóico, ?-caprolactona, ácido 6-amino-hexanóico, ?-caprolactama, hexametile-nodiamina, ácidos graxos lineares e álcoois graxos lineares que tem entre 7-25 carbonos de comprimento, alcanos lineares e alquenos lineares que são entre 6-24 carbonos de comprimento, ácido sebácico e ácido dodecanodióico compreendendo: a) converter um aldeído cn e piruvato a um intermediário de hidroxicetona cn+3 através da adição de aldol, e b) converter o intermediário de hidroxicetona cn+3 a compostos através de etapas enzimáticas, ou uma combinação de etapas enzimáticas e químicas. 1/1 high-yield route for producing compounds from renewable sources abstract Non-naturally occurring methods, compositions, and microbial organisms are provided here to prepare compounds such as 1-butanol, butyric acid, succinic acid, 1,4- butanediol, 1-pentanol, pentanoic acid, glutaric acid, 1,5-pentanodol, 1-hexanol, exanoic acid, adipic acid, 1,6-hexanediol, 6-hydroxyhexanoic acid, ?-caprolactone, 6-aminohexanoic acid , ?-caprolactam, hexamethylenediamine, linear fatty acids and linear fatty alcohols that are between 7-25 carbons in length, linear alkanes and linear alkenes that are between 6-24 carbons in length, sebacic acid and dodecanedioic acid comprising: a) converting a cn aldehyde and pyruvate to a cn+3 hydroxyketone intermediate through aldol addition, and b) converting the cn+3 hydroxyketone intermediate to compounds through enzymatic steps, or a combination of enzymatic and chemical steps. 1/1

Methods, cells and reagents for production of isoprene, derivatives and intermediates thereof

This application describes methods, including non-naturally occurring methods, for biosynthesizing 3-hydroxy~3-methylgSutaryl-coA and intermediates thereof, as well as non-naturally occurring hosts for producing 3-hydroxy-3-methyigiutaryl-coA. This application also describes methods, including non-naturally occurring methods, for biosynthesizing isoprene and intermediates thereof, as well as non-naturally occurring hosts for producing isoprene.

Genetically engineered bacterium with altered carbon monoxide dehydrogenase (codh) activity

The invention provides genetically engineered microorganisms with altered carbon monoxide dehydrogenase (CODH) activity and methods related thereto. In particular, the invention provides a genetically engineered carboxydotrophic acetogenic bacterium having decreased or eliminated activity of CODH1 and/or CODH2. In certain embodiments, the bacterium may also have increased activity of CODH/ACS. The invention further provides a method for producing a product by culturing the bacterium in the presence of a gaseous substrate comprising one or more of carbon monoxide, carbon dioxide, and hydrogen.

Fe-s fusion protein acting as electron transfer chain, novel carbon monoxide formate redox enzyme mediated through fes fusion protein, novel strain bcf12 derived from thermococcus wherein enzyme is transformed, and use thereof

The present invention relates to a Fe-S fusion protein acting as an electron transfer chain, a novel carbon monoxide formate redox enzyme mediated through a FeS fusion protein, a novel strain BCF12 derived from Thermococcus wherein an enzyme is transformed, and use thereof. The Fe-S fusion protein of the present invention, by mediating itself, physically directly binds two different enzymes, so that electrons generated in either of the enzymes can be directly transferred to the other enzyme through a Fe-S cluster of the Fe-S fusion protein. Thus, by directly supplying the electrons necessary for the production of a target substance, a high-efficiency production reaction of the target substance without leakage of electrons generated by either of the enzymes is possible. In addition, the present invention has the advantages that the overall enzyme reaction rate and yield can be significantly improved by using a new electron transfer reaction that did not previously exist, and the stability of each enzyme can also be ensured by allowing the enzymes to be physically fixed within a cell.

Genetically engineered bacterium comprising energy-generating fermentation pathway

The invention relates to a genetically engineered bacterium comprising an energy-generating fermentation pathway and methods related thereto. In particular, the invention provides a bacterium comprising a phosphate butyryltransferase (Ptb) and a butyrate kinase (Buk) (Ptb-Buk) that act on non-native substrates to produce a wide variety of products and intermediates. In certain embodiments, the invention relates to the introduction of Ptb-Buk into a C1-fixing microoorgansim capable of producing products from a gaseous substrate.

Fermentative production of 2-phenylethanol from gaseous substrates

Disclosed herein are methods for production of 2-phenylethanol by microbial fermentation of substrates comprising carbon monoxide and/or carbon dioxide and further disclosed are genetically modified microorganisms for use in such methods. Additionally, the processes disclosed herein are improved methods of 2-PE production that alleviate dependence on natural and petrochemical processes.

Method for converting non-ethanol producing, acetogenic strain to ethanol-producing strain and method for producing ethanol from same ethanol-producing strain by using carbon monoxide

The present invention relates to a transformed strain having ethanol production potential, constructed by introducing a foreign gene for ethanol production into a non-ethanol producing acetogen Eubacterium limosum and a method for producing ethanol, using the strain. According to the present invention, Eubacterium limosum which is a conventional acetogen lacking ethanol production potential is used to produce ethanol, which is a high value-added product, as a single product from carbon monoxide contained in waste gas.

Genetically engineered bacterium comprising energy-generating fermentation pathway

Genetically engineered bacterium with altered carbon monoxide dehydrogenase (codh) activity

The invention provides genetically engineered microorganisms with altered carbon monoxide dehydrogenase (CODH) activity and methods related thereto. In particular, the invention provides a genetically engineered carboxydotrophic acetogenic bacterium having decreased or eliminated activity of CODH1 and/or CODH2. In certain embodiments, the bacterium may also have increased activity of CODH/ACS. The invention further provides a method for producing a product by culturing the bacterium in the presence of a gaseous substrate comprising one or more of carbon monoxide, carbon dioxide, and hydrogen. The most suitable drawing: Figs. 1A-1D.

Genetically engineered bacteria, including energy-generating fermentation pathways

Corynebacterium glutamicum capable of producing L-proline in high yield and method for producing L-proline in high yield

本发明提供了一种生产L‑脯氨酸的谷氨酸棒杆菌，以及利用该菌株生产L‑脯氨酸的方法。本发明构建的生产L‑脯氨酸的谷氨酸棒杆菌，其中脯氨酸脱氢酶/吡咯‑5‑羧酸脱氢酶PutA失活，谷氨酸激酶ProB、谷氨酸‑5‑半醛脱氢酶ProA、吡咯‑5‑羧酸脱氢酶ProC、丙酮酸羧化酶Pyc、甘油醛‑3‑磷酸脱氢酶GapN、L‑脯氨酸外排蛋白ThrE或SerE活性增强，L‑谷氨酸外排蛋白MscCG失活，得到的菌株的L‑脯氨酸产量、转化率和生产强度较出发菌株显著提升，可降低L‑脯氨酸的生产成本。

Fermentative production of 2-phenylethanol from gaseous substrates

Disclosed herein are methods for production of 2-phenylethanol by microbial fermentation of substrates comprising carbon monoxide and/or carbon dioxide and further disclosed are genetically modified microorganisms for use in such methods. Additionally, the processes disclosed herein are improved methods of 2-PE production that alleviate dependence on natural and petrochemical processes.

Method for converting non-ethanol producing, acetogenic strain to ethanol-producing strain and method for producing ethanol from same ethanol-producing strain by using carbon monoxide

The present invention relates to a transformed strain having ethanol production potential, constructed by introducing a foreign gene for ethanol production into a non-ethanol producing acetogen Eubacterium limosum and a method for producing ethanol, using the strain. According to the present invention, Eubacterium limosum which is a conventional acetogen lacking ethanol production potential is used to produce ethanol, which is a high value-added product, as a single product from carbon monoxide contained in waste gas.

Recombinant microbial organisms exhibiting increased flux through fermentation pathways

Genetically engineered bacterium comprising energy-generating fermentation pathway

The invention relates to a genetically engineered bacterium comprising an energy-generating fermentation pathway and methods related thereto. In particular, the invention provides a bacterium comprising a phosphate butyryltransferase (Ptb) and a butyrate kinase (Buk) (Ptb-Buk) that act on non-native substrates to produce a wide variety of products and intermediates. In certain embodiments, the invention relates to the introduction of Ptb-Buk into a C1-fixing microoorgansim capable of producing products from a gaseous substrate.

Microbial organisms for converting acetyl-coa into crotyl alcohol and methods for producing crotyl alcohol

The present invention provides microorganisms capable of converting acetyl-coA into crotyl alcohol as well as fermentation methods for producing crotyl alcohol, either alone, or in combination with acetone and/or isopropanol. The microorganisms may be genetically engineered to express and/or disrupt one or more of the following enzymes: acetaldehyde dehydrogenase, alcohol dehydrogenase, bifunctional acetaldehyde/alcohol dehydrogenase, aldehyde oxidoreductase, phosphotransacetylase, acetate kinase, CoA-transferase A, CoA-transferase B, acetoacetate decarboxylase, secondary alcohol dehydrogenase, butyryl-CoA dehydrogenase (BCD), and/or trans-2-enoyl-CoA reductase (TER).

Noble Fe-S Fusion Protein Acting as an Electron Transfer Chain, and Use thereof

본 발명은 전자전달 기능을 하는 두 개 이상의 Fe-S 단백질을 연결한 Fe-S 융합단백질 및 이의 용도에 관한 것이다. 본 발명의 Fe-S 융합단백질은 이를 매개로 하여 서로 다른 두 개의 효소를 물리적으로 직접 결합함으로써, 어느 하나에서 생성된 전자가 Fe-S 융합단백질의 Fe-S 클러스터를 통해 다른 하나의 효소에 직접적으로 전달될 수 있고, 이에 따라. 목적 물질의 생산에 필요한 전자를 직접 공급함으로써 어느 하나의 효소에서 생성된 전자의 누출이 없는 고효율의 목적물질의 생산반응이 가능하다. 또한, 기존에 없던 새로운 전자전달 반응을 이용해 전체적인 효소 반응속도 및 수율을 획기적으로 개선할 수 있는 장점이 있고, 세포 내에서 효소들이 물리적으로 고정된 상태로 존재할 수 있도록 함으로써, 각 효소들의 안정성 또한 확보할 수 있다. The present invention relates to a Fe-S fusion protein linking two or more Fe-S proteins having an electron transport function and a use thereof. The Fe-S fusion protein of the present invention physically directly binds two different enzymes through this, so that an electron generated from one is directly connected to the other enzyme through the Fe-S cluster of the Fe-S fusion protein. Can be delivered to and accordingly. By directly supplying electrons required for the production of the target material, it is possible to produce a highly efficient target material without leakage of electrons generated by any one enzyme. In addition, it has the advantage of dramatically improving the overall enzyme reaction rate and yield by using a new electron transfer reaction that did not exist, and by allowing the enzymes to exist in a physically fixed state in the cell, the stability of each enzyme is also secured. can do.

RECOMBINANT MICROORGANISMS, MANIFESTING THE INCREASED FLOW DURING THE ENERGMENTAL WAY

Изобретение относится к рекомбинантной, карбоксидотрофной бактерии Clostridium, которая экспрессирует одну или более пируват:ферредоксин-оксидоредуктазу (ЕС 1.2.7.1), ацетолактатсинтазу (ЕС 2.2.1.6) и ацетолактат декарбоксилазу (ЕС 4.1.1.5). В изобретении дополнительно предложен способ получения продукта ферментации путем ферментации рекомбинантной бактерии в присутствии газообразного субстрата, содержащего СО, для получения одного или более из этанола, бутанола, изопропанола, изобутанола, высших спиртов, бутандиола, 2,3-бутандиола, сукцината, изопреноидов, жирных кислот, биополимеров и их смесей. The invention relates to a recombinant, carboxydotrophic bacterium Clostridium, which expresses one or more pyruvate: ferredoxin oxidoreductase (EC 1.2.7.1), acetolactate synthase (EC 2.2.1.6) and acetolactate decarboxylase (EC 4.1.1.5). The invention further provides a method for producing a fermentation product by fermenting a recombinant bacterium in the presence of a gaseous substrate containing CO to produce one or more of ethanol, butanol, isopropanol, isobutanol, higher alcohols, butanediol, 2,3-butanediol, succinate, isoprenoids, fatty acids, biopolymers and their mixtures.

Genetically engineered bacterium comprising energy-generating fermentation pathway

The invention relates to a genetically engineered bacterium comprising an energy-generating fermentation pathway and methods related thereto. In particular, the invention provides a bacterium comprising a phosphate butyryltransferase (Ptb) and a butyrate kinase (Buk) (Ptb-Buk) that act on non-native substrates to produce a wide variety of products and intermediates. In certain embodiments, the invention relates to the introduction of Ptb-Buk into a C1-fixing microoorgansim capable of producing products from a gaseous substrate.

Improvements in biological conversion and product recovery processes

The invention provides a process for reducing bio-catalytic oxidation of a product in a post-production stream. More particularly the invention provides a process for reducing bio-catalytic oxidation of an alcohol in a product stream, the product stream comprising an alcohol product, dissolved carbon dioxide, and at least one enzyme capable of oxidizing the alcohol. The invention finds applicability in fermentation processes, wherein a C1-fixing microorganism utilizes a C1-containing substrate to produce a fermentation product.

Recombinant microorganisms exhibiting increased flux through a fermentation pathway

Improvements in biological conversion and product recovery processes

本發明提供用於降低後期生產流中產物之生物催化性氧化的方法。更具體而言，本發明提供用於降低產物流中醇之生物催化性氧化的方法，所述產物流包括醇產物、溶解二氧化碳及至少一種能夠使所述醇氧化之酶。本發明適用於其中固定C1之微生物使用含有C1之受質以產生醱酵產物的醱酵製程。

Method for producing aldehyde

A method is described for producing an objective substance, for example, an aldehyde such as vanillin. The objective substance is produced from a carbon source or a precursor of the objective substance by using a microorganism having an ability to produce the objective substance, wherein the microorganism has been modified to have a specific carboxylic acid reductase (CAR) gene, such as a Gordonia CAR gene, Novosphingobium CAR gene, or Coccomyxa CAR gene.

Method for Converting Non-Ethanol Producing Acetogen to Ethanol Producing Acetogen and Method for Preparing Ethanol Using Carbon Monoxide from the Ethanol Producing Acetogen

에탄올 비생성 아세토젠인 Eubacterium limosum 에 에탄올 생산을 위한 외래 유전자를 도입하여 제작된 에탄올 생산능을 가지는 형질전환 균주 및 상기 균주를 이용한 에탄올의 제조방법에 관한 것으로, 본 발명에 따르면, 기존에 에탄올 생성능이 없는 아세토젠인 Eubacterium limosum 를 이용하여, 폐가스에 포함된 일산화탄소로부터 고부가가치의 에탄올을 단일 생산물로써 생산할 수 있다. Eubacterium , a non-ethanol-producing acetogen relates to a method of producing ethanol using the transformant strain, and the strain has a foreign production cost of ethanol produced by introducing a gene function for ethanol production in limosum, according to the present invention, the non-existing ethanol producing ability acetonitrile Zen Eubacterium limosum Using, it is possible to produce high value ethanol as a single product from carbon monoxide contained in the waste gas.

Genetically engineered bacterium comprising energy-generating fermentation pathway

The invention relates to a genetically engineered bacterium comprising an energy-generating fermentation pathway and methods related thereto. In particular, the invention provides a bacterium comprising a phosphate butyryltransferase (Ptb) and a butyrate kinase (Buk) (Ptb-Buk) that act on non-native substrates to produce a wide variety of products and intermediates. In certain embodiments, the invention relates to the introduction of Ptb-Buk into a Cl-fixing micr000rgansim capable of producing products from a gaseous substrate.

Improvements in biological conversion and product recovery processes

The invention provides a process for reducing bio-catalytic oxidation of a product in a post-production stream. More particularly the invention provides a process for reducing bio-catalytic oxidation of an alcohol in a product stream, the product stream comprising an alcohol product, dissolved carbon dioxide, and at least one enzyme capable of oxidizing the alcohol. The invention finds applicability in fermentation processes, wherein a C1-fixing microorganism utilizes a C1-containing substrate to produce a fermentation product.

Recombinant cell and production method for isoprene

本发明的目的在于提供能够由合成气等生产异戊二烯的一系列技术。本发明提供一种能够由C1化合物生产异戊二烯的重组细胞，其中，编码异戊二烯合成酶的核酸被导入具有基于非甲羟戊酸途径的异戊烯基二磷酸合成能力的宿主细胞中，该核酸在所述宿主细胞内进行表达，该C1化合物为选自由一氧化碳、二氧化碳、甲酸和甲醇组成的组中的至少一种。作为宿主细胞，可例示出梭菌(Clostridium)属细菌或穆尔氏菌(Moorella)属细菌。还提供使用了该重组细胞的异戊二烯的生产方法。

Method for converting non-ethanol producing, acetogenic strain to ethanol-producing strain and method for producing ethanol from same ethanol-producing strain by using carbon monoxide

The present invention relates to a transformed strain having ethanol production potential, constructed by introducing a foreign gene for ethanol production into a non-ethanol producing acetogen Eubacterium limosum and a method for producing ethanol, using the strain. According to the present invention, Eubacterium limosum which is a conventional acetogen lacking ethanol production potential is used to produce ethanol, which is a high value-added product, as a single product from carbon monoxide contained in waste gas.

Recombinant cell and production method for isoprene

본 발명은 합성 가스 등으로부터 이소프렌을 생산할 수 있는 일련의 기술을 제공하는 것을 목적으로 한다. 비메발론산 경로에 의한 이소펜테닐이인산 합성능을 갖는 숙주 세포에, 이소프렌 합성 효소를 코딩하는 핵산이 도입되어 이루어지고, 당해 핵산이 상기 숙주 세포 내에서 발현하며, 일산화탄소, 이산화탄소, 포름산 및 메탄올로 이루어지는 군에서 선택된 적어도 하나의 C1 화합물로부터 이소프렌을 생산 가능한 재조합 세포가 제공된다. 숙주 세포로서 클로스트리디움속 세균 또는 무렐라속 세균이 예시된다. 당해 재조합 세포를 사용한 이소프렌의 생산 방법도 제공된다.

Metabolic engineering for microbial production of terpenoid products

The invention relates to methods and bacterial strains for making terpene and terpenoid products, the bacterial strains having improved carbon pull through the MEP pathway and to a downstream recombinant synthesis pathway.

Genetically engineered bacterium comprising energy-generating fermentation pathway.

La presente invención se refiere a una bacteria diseñada por ingeniería genética que comprende una trayectoria de fermentación que genera energía y métodos relacionados a esta. En particular, la invención proporciona una bacteria que comprende una fosfato butiriltransferasa (Ptb) y una butirato cinasa (Buk) (Ptb-Buk) que actúa en sustratos no nativos para producir una amplia variedad de productos e intermediarios. En ciertas modalidades, la invención se refiere a la introducción de Ptb-Buk en un microorganismo que se fija a C1 capaz de producir productos a partir de un sustrato gaseoso.

Genetically engineered bacterium comprising energy-generating fermentation pathway.

La presente invención se refiere a una bacteria diseñada por ingeniería genética que comprende una trayectoria de fermentación que genera energía y métodos relacionados a esta. En particular, la invención proporciona una bacteria que comprende una fosfato butiriltransferasa (Ptb) y una butirato cinasa (Buk) (Ptb-Buk) que actúa en sustratos no nativos para producir una amplia variedad de productos e intermediarios. En ciertas modalidades, la invención se refiere a la introducción de Ptb-Buk en un microorganismo que se fija a C1 capaz de producir productos a partir de un sustrato gaseoso.

Synthetic growth on one-carbon substrates

多くのバイオテクノロジー関連生物は、安価で豊富な一炭素原料、例えば、ＣＯ２、ＣＯ、ホルムアルデヒド、メタノール、及びメタンを増殖に利用できず、代わりに糖などの複雑な原料を優先する。ホルミル－ＣｏＡ伸長経路を介して、生物が増殖及び維持のために１つの炭素分子を消費可能とする系を開示する。一炭素原料の利用は、バイオテクノロジー用途において生物を培養する主な手段としての糖の利用と置換できる。これにより、費用対効果が高くなり、論争を引き起こす原料としての食品の使用を回避し得る。ホルミル－ＣｏＡ伸長経路の中間体もまた、所望の化学産物に変換され得る。

Improved microbial production of fats

This invention describes a method of using microbial to produce fats, such as fatty acids and their derivatives, or products derived from the fatty acid synthesis cycle, such as hydroxyfatty acids, methyl ketones, and the like.

Microbial organisms for converting acetyl-coa into crotyl alcohol and methods for producing crotyl alcohol

The present invention provides microorganisms capable of converting acetyl-coA into crotyl alcohol as well as fermentation methods for producing crotyl alcohol, either alone, or in combination with acetone and/or isopropanol. The microorganisms may be genetically engineered to express and/or disrupt one or more of the following enzymes: acetaldehyde dehydrogenase, alcohol dehydrogenase, bifunctional acetaldehyde/alcohol dehydrogenase, aldehyde oxidoreductase, phosphotransacetylase, acetate kinase, CoA-transferase A, CoA-transferase B, acetoacetate decarboxylase, secondary alcohol dehydrogenase, butyryl-CoA dehydrogenase (BCD), and/or trans-2-enoyl-CoA reductase (TER).

Methods and compositions for the augmentation of pyruvate and acetyl-coa formation

The present disclosure identifies methods and compositions for modifying photoautotrophic organisms as hosts, such that the organisms efficiently convert carbon dioxide and light into pyruvate or acetyl-CoA, and in particular the use of such organisms for the commercial production of molecules derived from these precursors, e.g., ethanol.

Novel Strain from Thermococcus BCF12 and Method for Producing Formic Acid Using the Same

본 발명은 일산화탄소:포름산 산화·환원효소(carbon monoxide:formate oxidoreductase, CFOR) 유전자가 도입되어 포름산 생성능이 향상된 서모코커스 유래 신균주 BCF12 및 이를 이용한 포름산의 생산방법에 관한 것이다. 본 발명의 서모코커스 유래 신균주 BCF12는 Fe-S 단백질 유래 Fe-S 융합단백질을 매개로 하여 CO 탈수소효소(CO dehydrogenase, CODH)와 포름산 탈수소효소(formate dehydrogenase)를 물리적으로 직접 결합하여 CO 탈수소효소에서 생성된 전자가 Fe-S 융합단백질의 Fe-S 클러스터를 통해 포름산 탈수소효소에 직접적으로 전달되는 일산화탄소:포름산 산화·환원효소(carbon monoxide:formate oxidoreductase, CFOR)가 형질전환된 균주로, 기존에 없던 새로운 일산화탄소:포름산 산화·환원효소(carbon monoxide:formate oxidoreductase, CFOR) 반응을 이용해 포름산 생성의 반응속도 및 수율을 획기적으로 개선할 수 있는 장점이 있다. 또한, 세포 내에서 효소들이 물리적으로 고정된 상태로 존재할 수 있도록 함으로써, 각 효소들의 안정성 또한 확보할 수 있다. The present invention relates to a new strain BCF12 derived from Thermococcus having improved formic acid production ability by introducing a carbon monoxide:formate oxidoreductase (CFOR) gene and a method for producing formic acid using the same. The thermococcus-derived strain BCF12 of the present invention physically directly binds CO dehydrogenase (CO dehydrogenase, CODH) and formic acid dehydrogenase via a Fe-S fusion protein derived from Fe-S protein to form a CO dehydrogenase A carbon monoxide:formate oxidoreductase (CFOR) transformed by electrons generated directly from the Fe-S fusion protein to the formic acid dehydrogenase through a Fe-S cluster. The new carbon monoxide:formic acid oxidoreductase (CFOR) reaction has the advantage of dramatically improving the reaction rate and yield of formic acid production. In addition, by allowing enzymes to exist in a physically fixed state in a cell, stability of each enzyme can also be secured.

Recombinant cell and production method for isoprene

To provide a series of techniques capable of producing isoprene from syngas or the like. Provided is a recombinant cell prepared by introducing a nucleic acid encoding isoprene synthase into a host cell having an isopentenyl diphosphate synthesis ability by a non-mevalonate pathway, wherein the nucleic acid is expressed in the host cell, and the recombinant cell is capable of producing isoprene from at least one C1 compound selected from the group consisting of carbon monoxide, carbon dioxide, formic acid, and methanol. As the host cell, a Clostridium bacterium or a Moorella bacterium is exemplified. Also provided is a method for producing isoprene using the recombinant cell.

Genetically engineered bacterium comprising energy-generating fermentation pathway

The invention relates to a genetically engineered bacterium comprising an energy-generating fermentation pathway and methods related thereto. In particular, the invention provides a bacterium comprising a phosphate butyryltransferase (Ptb) and a butyrate kinase (Buk) (Ptb-Buk) that act on non-native substrates to produce a wide variety of products and intermediates. In certain embodiments, the invention relates to the introduction of Ptb-Buk into a Cl- fixing micr000rgansim capable of producing products from a gaseous substrate.

일산화탄소 이용능이 향상된 미생물 및 이의 용도

본 발명은 단백질 변이체, 이를 포함하는 CO 이용능이 향상된 미생물 및 이의 용도에 관한 것이다.

Genetically engineered bacterium comprising energy-generating fermentation pathway

The invention relates to a genetically engineered bacterium comprising an energy-generating fermentation pathway and methods related thereto. In particular, the invention provides a bacterium comprising a phosphate butyryltransferase (Ptb) and a butyrate kinase (Buk) (Ptb-Buk) that act on non-native substrates to produce a wide variety of products and intermediates. In certain embodiments, the invention relates to the introduction of Ptb-Buk into a Cl- fixing micr000rgansim capable of producing products from a gaseous substrate.

Genetically engineered bacterium comprising energy-generating fermentation pathway

The invention relates to a genetically engineered bacterium comprising an energy-generating fermentation pathway and methods related thereto. In particular, the invention provides a bacterium comprising a phosphate butyryltransferase (Ptb) and a butyrate kinase (Buk) (Ptb-Buk) that act on non-native substrates to produce a wide variety of products and intermediates. In certain embodiments, the invention relates to the introduction of Ptb-Buk into a Cl-fixing microoorgansim capable of producing products from a gaseous substrate. 99

Bacteria diseñada por ingeniería genética que comprende una trayectoria de fermentación que genera energía.

La presente invención se refiere a una bacteria diseñada por ingeniería genética que tiene una enzima que convierte 3-hidroxiisovaleril-CoA en 3-hidroxiisovalerato y una enzima que convierte 3-hidroxiisovalerato en isobutilen. Por lo general, la bacteria es capaz de producir isobutilen a partir de un sustrato gaseoso que contiene CO, CO2 y/o H2, como el syngas o un gas de desperdicio industrial.

Genetically engineered bacterium comprising energy-generating fermentation pathway

The invention relates to a genetically engineered bacterium comprising an energy-generating fermentation pathway and methods related thereto. In particular, the invention provides a bacterium comprising a phosphate butyryltransferase (Ptb) and a butyrate kinase (Buk) (Ptb-Buk) that act on non-native substrates to produce a wide variety of products and intermediates. In certain embodiments, the invention relates to the introduction of Ptb-Buk into a Cl-fixing microoorgansim capable of producing products from a gaseous substrate. 99

Recombinant microorganisms and uses therefor

Microorganisms are genetically engineered to produce 3-hydroxypropionate (3-HP). The microorganisms are carboxydotrophic acetogens. The microorganisms produce acetyl-coA using the Wood-Ljungdahl pathway for fixing CO/CO 2 . A β-alanine pyruvate aminotransferase from a microorganism that contains such an enzyme is introduced. Additionally, an acetyl-coA carboxylase may also be introduced. The production of 3-HP can be improved. This can be effected by improved promoters or higher copy number or enzymes that are catalytically more efficient.

Recombinant microorganisms and uses therefor