Nucleic acids and proteins and methods for making and using them

The invention provides polypeptides, including enzymes, structural proteins and binding proteins, polynucleotides encoding these polypeptides, and methods of making and using these polynucleotides and polypeptides. Polypeptides, including enzymes and antibodies, and nucleic acids of the invention can be used in industrial, experimental, food and feed processing, nutritional and pharmaceutical applications, e.g., for food and feed supplements, colorants, neutraceuticals, cosmetic and pharmaceutical needs.

Genetically engineered bacteria synthesizing styrene taking biomass hydrolysate as raw material and construction method and application thereof

METHOD FOR PRODUCING FRUCTOSE-6-PHOSPHATE FROM DIHYDROXYACETONE PHOSPHATE AND GLYCERALDEHYDE-3-PHOSPHATE

Described is a method for the production of fructose-6-phosphate (F6P) from dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (G3P) comprising the steps of: (a) enzymatically converting dihydroxyacetone phosphate (DHAP) into dihydroxyacetone (DHA); and (b) enzymatically converting the thus produced dihydroxyacetone (DHA) and glyceraldehyde-3-phosphate (G3P) into fructose-6-phosphate (F6P); or comprising the steps of: (a') enzymatically converting glyceraldehyde-3-phosphate (G3P) into glyceraldehyde; and (b') enzymatically converting the thus produced glyceraldehyde together with dihydroxyacetone phosphate (DHAP) into fructose-1-phosphate (F1P); and (c') enzymatically converting the thus produced fructose-1-phosphate (F1P) into fructose-6-phosphate (F6P).

METABOLIC ENGINEERING OF THE SHIKIMATE PATHWAY

The present disclosure relates to engineered microorganisms that produce amino acids and amino acid intermediates. In particular, the disclosure relates to recombinant nucleic acids encoding operons that increase production of aromatic amino acids and the aromatic amino acid intermediate shikimate; microorganisms with increased production of aromatic amino acids and the aromatic amino acid intermediate shikimate; and methods related to the production of aromatic amino acids, the aromatic amino acid intermediate shikimate, and commodity chemicals derived therefrom.

Nucleic acids and proteins and methods for making and using them

Fusion proteins comprising an aldolase enzyme joined to a maltose binding protein

The present invention refers to an enzyme consisting of a fusion protein particularly useful as shown through-out the present invention for carrying out the carbon-carbon bond-forming reaction known as the aldol Reaction, preferably for carrying out an aldol reaction by using aldehydes as substrates and preferably pyruvate or a salt thereof, for producing hydroxyketoacids. Said enzyme is made by binding an aldolase to a maltose binding protein. The enzymes display full activity under “highly denaturing” substrate loadings (aldehydes, >1 M).

METABOLIC ENGINEERING OF THE SHIKIMATE PATHWAY

The present disclosure relates to engineered microorganisms that produce amino acids and amino acid intermediates. In particular, the disclosure relates to recombinant nucleic acids encoding operons that increase production of aromatic amino acids and the aromatic amino acid intermediate shikimate; microorganisms with increased production of aromatic amino acids and the aromatic amino acid intermediate shikimate; and methods related to the production of aromatic amino acids, the aromatic amino acid intermediate shikimate, and commodity chemicals derived therefrom.

Method for producing fructose-6-phosphate from dihydroxy acetone phosphate and glyceraldehyde-3-phosphate

Described is a method for the production of fructose-6-phosphate (F6P) from dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (G3P) comprising the steps of: (a) enzymatically converting dihydroxyacetone phosphate (DHAP) into dihydroxyacetone (DHA); and (b) enzymatically converting the thus produced dihydroxyacetone (DHA) and glyceraldehyde-3-phosphate (G3P) into fructose-6-phosphate (F6P); or comprising the steps of: (a′) enzymatically converting glyceraldehyde-3-phosphate (G3P) into glyceraldehyde; and (b′) enzymatically converting the thus produced glyceraldehyde together with dihydroxyacetone phosphate (DHAP) into fructose-1-phosphate (F1P); and (c′) enzymatically converting the thus produced fructose-1-phosphate (F1P) into fructose-6-phosphate (F6P).

COMPOSITIONS, SYSTEMS, AND METHODS FOR ARTIFICIAL CARBON FIXATION, CHEMICAL SYNTHESIS, AND/OR PRODUCTION OF USEFUL PRODUCTS

Construction of escherichia coli genetically engineered bacterium and application of escherichia coli genetically engineered bacterium to production of L-tryptophan

(R)-selective nitroaldol reaction catalysed by proteins of the cupin superfamily

The present invention relates to a method for producing chiral β-nitro alcohol compounds. The invention relates in particular to an (R)-selective cupin-nitroaldolase, which enantioselectively can catalyze the Henry reaction, wherein an aldehyde or ketone compound is converted to the corresponding β-nitro alcohol compound in the presence of a nitroalkane compound and a cupin-nitroaldolase.

Metabolic engineering of the shikimate pathway

The present disclosure relates to engineered microorganisms that produce amino acids and amino acid intermediates. In particular, the disclosure relates to recombinant nucleic acids encoding operons that increase production of aromatic amino acids and the aromatic amino acid intermediate shikimate; microorganisms with increased production of aromatic amino acids and the aromatic amino acid intermediate shikimate; and methods related to the production of aromatic amino acids, the aromatic amino acid intermediate shikimate, and commodity chemicals derived therefrom.

METHOD FOR PRODUCING FRUCTOSE-6-PHOSPHATE FROM DIHYDROXYACETONE PHOSPHATE AND GLYCERALDEHYDE-3-PHOSPHATE

Described is a method for the production of fructose-6-phosphate (F6P) from dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (G3P) comprising the steps of: (a) enzymatically converting dihydroxyacetone phosphate (DHAP) into dihydroxyacetone (DHA); and (b) enzymatically converting the thus produced dihydroxyacetone (DHA) and glyceraldehyde-3-phosphate (G3P) into fructose-6-phosphate (F6P); or comprising the steps of: (a′) enzymatically converting glyceraldehyde-3-phosphate (G3P) into glyceraldehyde; and (b′) enzymatically converting the thus produced glyceraldehyde together with dihydroxyacetone phosphate (DHAP) into fructose-1-phosphate (F1P); and (c′) enzymatically converting the thus produced fructose-1-phosphate (F1P) into fructose-6-phosphate (F6P).

Process for preparing acetoin using methanotrophs or transformant thereof

Method for mass producing human blood coagulation factor vii derivative

Recombinant Escherichia coli with high yield of L-threonine and construction method thereof

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 ...

(R)-SELECTIVE NITROALDOL REACTION CATALYSED BY PROTEINS OF THE CUPIN SUPERFAMILY

The present invention relates to a method for producing chiral β-nitro alcohol compounds. The invention relates in particular to an (R)-selective cupin-nitroaldolase, which enantioselectively can catalyze the Henry reaction, wherein an aldehyde or ketone compound is converted to the corresponding β-nitro alcohol compound in the presence of a nitroalkane compound and a cupin-nitroaldolase.

Recombinant bacterium for producing glycolic acid by using xylose as well as construction method and application thereof

NUCLEIC ACIDS AND PROTEINS AND METHODS FOR MAKING AND USING THEM

The invention provides polypeptides, including enzymes, structural proteins and binding proteins, polynucleotides encoding these polypeptides, and methods of making and using these polynucleotides and polypeptides. Polypeptides, including enzymes and antibodies, and nucleic acids of the invention can be used in industrial, experimental, food and feed processing, nutritional and pharmaceutical applications, e.g., for food and feed supplements, colorants, neutraceuticals, cosmetic and pharmaceutical needs. 1. An isolated or recombinant nucleic acid comprising(a) a nucleic acid sequence having at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63% 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, and all nucleic acids disclosed in the SEQ ID listing, which include all odd numbered SEQ ID NOs: from SEQ ID NO:1 through SEQ ID NO:26,897, over a region of at least about 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150 or more residues,wherein the nucleic acid encodes at least one polypeptide having an enzymatic activity, or encodes a polypeptide or peptide capable of generating an antibody that binds specifically to a polypeptide having a sequence comprising any of the even numbered SEQ ID NOs: in the sequence listing, including from SEQ ID NO:2 through SEQ ID NO:26,898;(b) a nucleic acid sequence that hybridizes under stringent conditions to a nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, and all ...

ORGANISMS PRODUCING LESS CROTONIC ACID

The present invention relates to a recombinant organism or microorganism having a decreased pool of crotonic acid compared to the organism or microorganism from which it is derived due to at least: (i) a decreased conversion of acetaldehyde into crotonaldehyde; and/or (ii) a decreased conversion of crotonyl-CoA into crotonaldehyde; and/or (iii) a decreased conversion of crotonaldehyde into crotonic acid. Moreover, the present invention relates to the use of such a recombinant organism or microorganism for the production of alkenes with the enzyme ferulic acid decarboxylase. Further, the present invention relates to a method for the production of isobutene or butadiene by culturing such a recombinant organism or microorganism in a suitable culture medium under suitable conditions.

Nucleic acids and proteins and methods for making and using them

The invention provides polypeptides, including enzymes, structural proteins and binding proteins, polynucleotides encoding these polypeptides, and methods of making and using these polynucleotides and polypeptides. Polypeptides, including enzymes and antibodies, and nucleic acids of the invention can be used in industrial, experimental, food and feed processing, nutritional and pharmaceutical applications, e.g., for food and feed supplements, colorants, neutraceuticals, cosmetic and pharmaceutical needs.

A HIGH YIELD ROUTE FOR THE PRODUCTION OF COMPOUNDS FROM RENEWABLE SOURCES

método para produzir frutose-6-fosfato, micro-organismo recombinante, combinação de enzimas, composição e uso do micro-organismo

Nucleic Acids and Proteins and Methods for Making and Using Them

The invention provides polypeptides, including enzymes, structural proteins and binding proteins, polynucleotides encoding these polypeptides, and methods of making and using these polynucleotides and polypeptides. Polypeptides, including enzymes and antibodies, and nucleic acids of the invention can be used in industrial, experimental, food and feed processing, nutritional and pharmaceutical applications, e.g., for food and feed supplements, colorants, neutraceuticals, cosmetic and pharmaceutical needs.

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 ...

produção de ácido 2-ceto-3-desoxi-d-glucônico em fungos filamentosos

FUSION PROTEINS COMPRISING AN ALDOLASE ENZYME JOINED TO A MALTOSE BINDING PROTEIN

The present invention refers to an enzyme consisting of a fusion protein particularly useful as shown through-out the present invention for carrying out the carbon-carbon bond-forming reaction known as the aldol Reaction, preferably for carrying out an aldol reaction by using aldehydes as substrates and preferably pyruvate or a salt thereof, for producing hydroxyketoacids. Said enzyme is made by binding an aldolase to a maltose binding protein. The enzymes display full activity under “highly denaturing” substrate loadings (aldehydes, >1 M). 1. A composition comprising a fusion protein which in turn comprises a 2-keto-3-deoxy--rhamnonate aldolase or a variant thereof , wherein the term “variant” is understood as a protein exhibiting 2-keto-3-deoxy--rhamnonate aldolase activity and at least 80% sequence identity with amino acid sequence SEQ ID NO 1 or with an amino acid sequence coded by SEQ ID NO 2 bound to , optionally through a peptide linker , a maltose binding protein (MBP).2. The composition of claim 1 , wherein said aldolase is bound to the MBP through a peptide linker having from 3 to 50 amino acids in length.3. The composition of or claim 1 , wherein the MBP forming part of the fusion protein is the MBP of SEQ ID NO 8 or a variant thereof claim 1 , wherein the term “variant” is understood as a protein exhibiting at least 80% sequence identity with the MBP having amino acid sequence SEQ ID NO 8.4. The composition of claim 3 , wherein the aldolase enzyme forming part of the fusion protein is the 2-keto-3-deoxy--rhamnonate aldolase consisting of SEQ ID NO 1 and the MBP forming part of the fusion protein is the MBP of SEQ ID NO 8.5. The composition of any of to claim 3 , wherein said composition further comprises any of the following components: protein divalent metals claim 3 , preferably Mg claim 3 , Co and/or Ni claim 3 , additional enzymes such as reductases claim 3 , decarboxylases or transaminases such as the transaminase Prozomix TA051 (preferably in the ...

Method for the incorporation of formaldehyde into biomass

The present disclosure relates to a method for the incorporation of formaldehyde into biomass comprising the following enzymatically catalyzed steps: (1) condensation of pyruvate with formaldehyde into 4-hydroxy-2-oxobutanoic acid (HOB); (2) amination of the thus produced 4-hydroxy-2-oxobutanoic acid (HOB) to produce homoserine; (3) conversion of thus produced homoserine to threonine; (4) conversion of the thus produced threonine into glycine and acetaldehyde or acetyl-CoA; (5) condensation of the thus produced glycine with formaldehyde to produce serine; and (6) conversion of the thus produced serine to produce pyruvate, wherein said pyruvate can then be used as a substrate in step (1). The disclosure also relates to enzymes for catalyzing the corresponding enzymatic reactions and recombinant microorganisms which express the enzymes for catalyzing the corresponding enzymatic reactions.

Nucleic Acids and Proteins and Methods for Making and Using Them

The invention provides polypeptides, including enzymes, structural proteins and binding proteins, polynucleotides encoding these polypeptides, and methods of making and using these polynucleotides and polypeptides. Polypeptides, including enzymes and antibodies, and nucleic acids of the invention can be used in industrial, experimental, food and feed processing, nutritional and pharmaceutical applications, e.g., for food and feed supplements, colorants, neutraceuticals, cosmetic and pharmaceutical needs.

(r)-selective nitroaldol reaction catalysed by proteins of the cupin superfamily

(R)-SELECTIVE NITROALDOL REACTION CATALYSED BY PROTEINS OF THE CUPIN SUPERFAMILY

The present invention relates to a method for producing chiral β-nitro alcohol compounds. The invention relates in particular to an (R)-selective cupin-nitroaldolase, which enantioselectively can catalyze the Henry reaction, wherein an aldehyde or ketone compound is converted to the corresponding β-nitro alcohol compound in the presence of a nitroalkane compound and a cupin-nitroaldolase. 121.-. (canceled)22. A cupin-nitroaldolase variant protein characterized in that it comprises the amino acid sequence of the general formula:{'br': None, '(X1)(X2)(X3)(X4)F(X5)PGAR(X6)(X7)WH(X8)HP(X9)G, wherein'}X1 is an A, V, L, F, Y, M, S, T, G, H, N, K, or R residue;X2 is any amino acid;X3 is a V, A, I, C, M, H, or T residue;X4 is any amino acid;X5 is any amino acid;X6 is a T, S or N residue;X7: is any amino acid;X8: is a T, S, or I residues;X9: is any amino acid;and wherein at least one of positions X1, or X3 is substituted by a H, K, R or T residue.23. The cupin-nitroaldolase variant protein according to claim 22 , wherein in the general characterized in that it comprises the amino acid sequence of the general formula{'br': None, '(X1)(X2)(X3)(X4)F(X5)PGAR(X6)(X7)WH(X8)HP(X9)G,'}X1 is an A or N residue;X2 is an S, H, A or T residue;X3 is a V residue;X4 is a T or R residue;X5 is an E residue;X6 is a T residue;X7: is an A residue;X8: is a T residue;X9: is an L residue;and wherein at least one of positions X1, or X3 is substituted by a H, K, R or T residue.24. The cupin-nitroaldolase variant protein according to claim 22 , wherein the cupin-nitroaldolase is at least 85% identical to the respective wild type enzyme.25. The cupin-nitroaldolase variant protein according to claim 23 , wherein the cupin-nitroaldolase is at least 85% identical to the respective wild type enzyme.26. The cupin-nitroaldolase variant protein according to having one or more of the following mutations: A40H claim 22 , A40R claim 22 , V42T and/or Q110H according to the amino acid numbering of SEQ ID NO: 1 or ...

ENGINEERED DEOXYRIBOSE-PHOSPHATE ALDOLASES

The present invention provides engineered deoxyribose-phosphate aldolase polypeptides useful under industrial process conditions for the production of pharmaceutical and fine chemical compounds. 1. An engineered deoxyribose-phosphate aldolase comprising a polypeptide sequence having at least 85% , 86% , 87% , 88% , 89% , 90% , 91% , 92% , 93% , 94% , 95% , 96% , 97% , 98% , 99% , or more sequence identity to SEQ ID NOS: 2 , 6 , and/or 466 , or a functional fragment thereof , wherein said engineered deoxyribose-phosphate aldolase comprises at least one substitution or substitution set in said polypeptide sequence , and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 2 , 6 , and/or 466.2. The engineered deoxyribose-phosphate aldolase of claim 1 , wherein said polypeptide sequence has at least 85% claim 1 , 86% claim 1 , 87% claim 1 , 88% claim 1 , 89% claim 1 , 90% claim 1 , 91% claim 1 , 92% claim 1 , 93% claim 1 , 94% claim 1 , 95% claim 1 , 96% claim 1 , 97% claim 1 , 98% claim 1 , 99% claim 1 , or more sequence identity to SEQ ID NO:2 claim 1 , wherein said engineered deoxyribose-phosphate aldolase comprises at least one substitution or substitution set in said polypeptide sequence at one or more positions selected from 10/47/66/141/145/156 claim 1 , 2 claim 1 , 6 claim 1 , 9 claim 1 , 10/47/88/156 claim 1 , 13 claim 1 , 31 claim 1 , 46 claim 1 , 47 claim 1 , 47/134/141/212 claim 1 , 66 claim 1 , 66/88/112/134/141/143/145/212 claim 1 , 71 claim 1 , 72 claim 1 , 88 claim 1 , 94 claim 1 , 102 claim 1 , 104 claim 1 , 112 claim 1 , 116 claim 1 , 133 claim 1 , 133/173/204/235/236 claim 1 , 134 claim 1 , 145 claim 1 , 145/173 claim 1 , 147 claim 1 , 173 claim 1 , 184 claim 1 , 189 claim 1 , 197 claim 1 , 203 claim 1 , 204 claim 1 , 207 claim 1 , 226 claim 1 , 235 claim 1 , 235/236 claim 1 , and 236 claim 1 , and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: ...

NON-CO2 EVOLVING METABOLIC PATHWAY FOR CHEMICAL PRODUCTION

Provided are microorganisms that catalyze the synthesis of chemicals and biochemicals from a suitable carbon source. Also provided are methods of generating such organisms and methods of synthesizing chemicals and biochemicals using such organisms. 1. A recombinant microorganism comprising a non-COevolving metabolic pathway for the synthesis of acetyl phosphate with improved carbon yield beyond 1:2 molar ratio (fructose 6-phosphate:Acetyl phosphate) from a carbon substrate using a pathway comprising an enzyme having (i) fructose-6-phosphoketolase (Fpk) activity and/or xylulose-5-phosphoketolase (Xpk) activity and (ii) a fructose 1 ,6 bisphosphatase or a sedoheptuloase 1 ,6 bisphosphatase activity.2. The recombinant microorganism of claim 1 , wherein the microorganism can convert a sugar phosphate to acetyl phosphate with improved yield beyond those obtained by pathways that involve pyruvate decarboxylation.3. The recombinant microorganism of claim 2 , wherein the sugar phosphate is selected from the group consisting of: sugar phosphates of a triose claim 2 , an erythrose claim 2 , a pentose claim 2 , a hexose claim 2 , and a sedoheptulose.4. The recombinant microorganism of claim 3 , wherein the sugar phosphate of a triose is selected from the group consisting of G3P and DHAP; wherein the pentose is selected from the group consisting of RSP claim 3 , Ru5P claim 3 , RuBP claim 3 , and X5P; wherein the hexose is selected from the group consisting of F6P claim 3 , H6P claim 3 , FBP claim 3 , and G6P; and wherein the sedoheptulose is selected from the group consisting of S7P and SBP.5. The recombinant microorganism of claim 3 , wherein the sugar phosphates are derived from a carbon source selected from the group consisting of methanol claim 3 , methane claim 3 , CO claim 3 , CO claim 3 , formaldehyde claim 3 , formate claim 3 , glycerol claim 3 , a carbohydrate having the general formula CHO claim 3 , wherein n=3 to 7 claim 3 , and cellulose.6. The recombinant ...

ALDOLASE, ALDOLASE MUTANT, AND METHOD AND COMPOSITION FOR PRODUCING TAGATOSE BY USING SAME

There are provided aldolase, an aldolase mutant, a method for producing tagatose, and a composition for producing tagatose using the same. The technical feature of the present invention is environment-friendly due to the use of an enzyme acquired from microorganisms, requires only a simple process of enzyme-immobilization, uses a low-cost substrate compared with that of a conventional method for producing tagatose, and has a remarkably high yield, thereby greatly reducing production cost while maximizing production effect. 1. A method of producing tagatose from fructose , the method comprising:reacting fructose 6-phosphate with tagatose 6-phosphate epimerase or a mutant thereof to obtain tagatose 6-phosphate; andconverting the tagatose 6-phosphate to tagatose.2. The method of claim 1 , wherein the tagatose 6-phosphate epimerase catalyzes the conversion of fructose 6-phosphate to tagatose-6-phosphate.3. The method of claim 1 , wherein the tagatose 6-phosphate epimerase is SEQ ID NOS: 1.4. The method of claim 1 , wherein the tagatose 6-phosphate epimerase is SEQ ID NOS: 2.5. The method of claim 1 , wherein the tagatose 6-phosphate epimerase is SEQ ID NOS: 3.6. The method of claim 1 , wherein the tagatose 6-phosphate epimerase is SEQ ID NOS: 4.7. The method of claim 1 , wherein the mutant comprises at least one amino acid substitution of SEQ ID NO: 1 selected from the group consisting of:i) a substitution of arginine residue with glutamine at a position corresponding to position 332,ii) a substitution of glutamine residue with alanine at a position corresponding to position 314,iii) a substitution of histidine residue with alanine at a position corresponding to position 227, andiv) a substitution of serine residue with alanine at a position corresponding to position 62.8. The method of claim 1 , wherein the fructose 6-phosphate is obtained by treating fructose or a fructose-containing material with hexokinase.9. The method of claim 1 , wherein the tagatose is obtained ...

MICROORGANISMS AND METHODS FOR THE CO-PRODUCTION OF ETHYLENE GLYCOL AND THREE CARBON COMPOUNDS

The present application relates to recombinant microorganisms useful in the biosynthesis of monoethylene glycol (MEG) and one or more three-carbon compounds such as acetone, isopropanol or propene. The MEG and one or more three-carbon compounds described herein are useful as starting material for production of other compounds or as end products for industrial and household use. The application further relates to recombinant microorganisms co-expressing a C2 branch pathway and a C3 branch pathway for the production of MEG and one or more three-carbon compounds. Also provided are methods of producing MEG and one or more three-carbon compounds using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or optionally the products MEG and one or more three-carbon compounds. 2. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises at least one endogenous or exogenous nucleic acid molecule encoding a secondary alcohol dehydrogenase that catalyzes the conversion of acetone to isopropanol.3. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises: at least one endogenous or exogenous nucleic acid molecule encoding a secondary alcohol dehydrogenase that catalyzes the conversion of acetone to isopropanol; and at least one endogenous or exogenous nucleic acid molecule encoding a dehydratase that catalyzes the conversion of isopropanol to propene.4. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises one or more modifications selected from the group consisting of:(a) a deletion, insertion, or loss of function mutation in a gene encoding a D-xylose isomerase that catalyzes the conversion of D-xylose to D-xylulose;(b) a deletion, insertion, or loss of function mutation in a gene encoding a glycolaldehyde dehydrogenase that catalyzes the conversion of glycolaldehyde to glycolic acid; and(c) a deletion, ...

IMPROVED GLYCEROL FREE ETHANOL PRODUCTION

The invention relates to a recombinant cell, preferably a yeast cell comprising one or more genes coding for an enzyme having glycerol dehydrogenase activity, one or more genes coding dihydroxyacetone kinase (E.C. 2.7.1.28 and/or E.C. 2.7.1.29); one or more genes coding for an enzyme in an acetyl-CoA-production pathway and one or more genes coding for an enzyme having at least NAD dependent acetylating acetaldehyde dehydrogenase activity (EC 1.2.1.10 or EC 1.1.1.2), and optionally one or more genes coding for a glycerol transporter. This cell can be used for the production of ethanol and advantageously produces little or no glycerol. 1. A recombinant cell , optionally a yeast cell , said recombinant cell comprising:one or more genes coding for an enzyme having glycerol dehydrogenase activity;one or more genes coding dihydroxyacetone kinase (E.C. 2.7.1.28 and/or E.C. 2.7.1.29);one or more genes coding for an enzyme in an acetyl-CoA-production pathway; and{'sup': '+', 'one or more genes coding for an enzyme having at least NAD dependent acetylating acetaldehyde dehydrogenase activity (EC 1.2.1.10 or EC 1.1.1.2); and optionally'}one or more genes coding for a glycerol transporter.2. The Cell according to wherein the enzyme having glycerol dehydrogenase activity is a NAD linked glycerol dehydrogenase (EC 1.1.1.6).3. The Cell according to wherein the enzyme having glycerol dehydrogenase activity is a NADP linked glycerol dehydrogenase (EC 1.1.1.72).4. The recombinant cell according to wherein the one or more genes coding for an enzyme in an acetyl-CoA-production pathway comprises:one or more genes coding for an enzyme having phosphoketolase (PKL) activity (EC 4.1.2.9 or EC 4.1.2.22) or an enzyme having an amino acid sequence according SEQ ID NO: 5, 6, 7, or 8, or functional homologues thereof having a sequence identity of at least 50%, and/orone or more genes coding for an enzyme having phosphotransacetylase (PTA) activity (EC 2.3.1.8) or an enzyme having an amino acid ...

Composition for producing tagatose and method of producing tagatose using the same

Provided are a composition for producing tagatose, comprising fructose-4-epimerase, and a method of producing tagatose using the same.

Polypeptide Assemblies and Methods for the Production Thereof

The application discloses multimeric assemblies including multiple oligomeric substructures, where each oligomeric substructure includes multiple proteins that self-interact around at least one axis of rotational symmetry, where each protein includes one or more polypeptide-polypeptide interface (“O interface”); and one or more polypeptide domain that is capable of effecting membrane scission and release of an enveloped multimeric assembly from a cell by recruiting the ESCRT machinery to the site of budding by binding to one or more proteins in the eukaryotic ESCRT complex (“L domain”); and where the multimeric assembly includes one or more subunits comprising one or more polypeptide domain that is capable of interacting with a lipid bilayer (“M domain”), as well as membrane-enveloped versions of the multimeric assemblies. 1. A multimeric assembly , comprising a plurality of oligomeric substructures , wherein each oligomeric substructure comprises a plurality of proteins that self-interact around at least one axis of rotational symmetry , wherein each protein comprises:(a) one or more polypeptide-polypeptide interface (“O interface”);(b) one or more polypeptide domain that is capable of effecting membrane scission and release of an enveloped multimeric assembly from a cell by recruiting the ESCRT machinery to the site of budding by binding directly or indirectly to one or more ESCRT or ESCRT-associated proteins (“L domain”);wherein the multimeric assembly comprises one or more polypeptide domain that is capable of interacting with a lipid bilayer (“M domain”);wherein the M domain, L domain, and O interface are not each present in a single naturally occurring protein, wherein the plurality of oligomeric substructures interact with each other at the one or more O interfaces.2. The multimeric assembly of claim 1 , wherein each oligomeric structure comprises one or more M domain claim 1 , or wherein each protein comprises one or more M domain.3. The multimeric assembly ...

Electrochemical Bioreactor Module and Engineered Metabolic Pathways for 1-Butanol Production with High Carbon Efficiency

A combination of an electrochemical device for delivering reducing equivalents to a cell, and engineered metabolic pathways within the cell capable of utilizing the electrochemically provided reducing equivalents is disclosed. Such a combination allows the production of commodity chemicals by fermentation to proceed with increased carbon efficiency. 1. A system for 1-butanol production , comprising:an electrochemical bioreactor module for providing reducing equivalents;a first engineered pathway for producing 1-butanol from acetyl-CoA; anda second engineered pathway for recovering carbon as formate from pyruvate, and converting the recovered formate to fructose-6-phosphate;wherein the reducing equivalents are provided to one or more redox enzymes in the first and/or second engineered pathways; and wherein optionally the first and second engineered pathways are present in an engineered cell.2. The system of wherein the first engineered pathway comprises acetyl-CoA acetyltransferase (AtoB claim 1 , EC 2.3.1.9) claim 1 , 3-hydroxybutyryl-CoA dehydrogenase (Hbd claim 1 , EC 1.1.1.157) claim 1 , 3-hydroxybutyryl-CoA dehydratase (Crt claim 1 , EC 4.2.1.5) claim 1 , trans-enoyl-CoA reductase (Ter claim 1 , EC 1.3.1.38) and aldehyde/alcohol dehydrogenase (AdhE2 claim 1 , EC 1.2.157/EC 1.1.1.1).3. The system of or wherein the second engineered pathway comprises pyruvate:formate lyase (Pfl claim 1 , EC 2.3.1.54) claim 1 , formaldehyde dehydrogenase (Fld claim 1 , EC 1.2.1.46) claim 1 , hexulose-6-phosphate synthase (HPS claim 1 , EC 4.1.2.43) claim 1 , and 6-phospho-3-hexuloisomerase (HPI claim 1 , EC 5.3.1.27).4. The system of or wherein in the engineered cell claim 1 , the endogenous pyruvate dehydrogenase (Pdh claim 1 , EC 1.2.4.1) has been disabled claim 1 , deleted or otherwise rendered non-functional.5. The system of or wherein in the engineered cell claim 1 , the endogenous fumarate reductase (FrdBC claim 1 , EC 1.3.1.6) claim 1 , lactate dehydrogenase (Ldh claim 1 , ...

ANAEROBIC FERMENTATIVE PRODUCTION OF FURANDICARBOXYLIC ACID

The present disclosure provides recombinant microorganisms and methods for the anaerobic production of 2,4-furandicarboxylic acid from one or more carbon sources. The microorganisms and methods provide redox-balanced and ATP positive pathways for co-producing 2,4-furandicarboxylic acid with ethanol and for co-producing 2,4-furandicarboxylic acid with ethanol and 1-propanol. The method provides recombinant microorganisms that express endogenous and/or exogenous nucleic acid molecules encoding polypeptides that catalyze the conversion of a carbon source into 2,4-furandicarboxylic acid and that coupled the 2,4-furandicarboxylic acid pathway with an additional metabolic pathway.

Biosynthetic methods and systems for producing monosaccharides

The present disclosure is related to biosynthetic methods of forming monosaccharides, and systems for generating the same. A benefit of the methods and systems disclosed herein can include the sustainable production of monosaccharides in an automated process. A benefit of the methods and systems herein can be the generation of monosaccharides from renewable source materials. An additional benefit of the methods and systems herein can include the use of abundant feedstocks, such as carbon dioxide, for the efficient generation of select monosaccharides for use as nutrients and for other useful applications. Another benefit of the methods and systems disclosed herein can include reduction of excess carbon dioxide from the environment.

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

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

CULTURE MODIFIED TO CONVERT METHANE OR METHANOL TO 3-HYDROXYPROPRIONATE

Provided are engineered organisms which can convert methane or methanol to 3-hydroxypropionate. 1. A synthetic culture comprising one or more microorganisms comprising one or more modifications that improve the production of a product from a substrate , wherein the substrate comprises methane and/or methanol.2. The synthetic culture according to claim 1 , wherein the substrate comprises methane.3. The synthetic culture according to claim 2 , wherein the product comprises 3-hydroxyproprionate.4. The synthetic culture according to claim 1 , wherein the product comprises 3-hydroxyproprionate.5. The synthetic culture according to claim 1 , wherein the product comprises a substance derived from acetyl-CoA and/or malonyl-CoA.6Escherichia coli.. The synthetic culture according to claim 1 , wherein at least one of the one or more microorganisms comprises7. The synthetic culture according to claim 1 , wherein the one or more microorganisms comprises a first at least one microorganism and a second at least one microorganism claim 1 , wherein the first at least one microorganism produces methanol from methane and the second at least one microorganism produces 3-hydroxypropionate from methanol.8. The synthetic culture according to claim 1 , wherein the one or more modifications comprise exogenous polynucleotides or deletion of one or more genes.9. The synthetic culture according to claim 8 , wherein the exogenous polynucleotides encode polypeptides selected from one or more polypeptides comprising methane monooxygenase (EC 1.14.13.25) claim 8 , malonyl-CoA reductase (EC 1.2.1.75) claim 8 , acetyl-CoA carboxylase (EC 6.4.1.2) claim 8 , methanol dehydrogenase (EC 1.1.1.244 or EC 1.1.2.7) claim 8 , 3-hexulose-6-phosphate synthase (EC 4.1.2.43) claim 8 , and/or 6-phospho-3-hexuloisomerase (EC 5.3.1.27).10Bacillus methanolicus, Bacillus stearothermophilusCorynebacterium glutamicum.. The synthetic culture according to claim 9 , wherein the methanol dehydrogenase comprises a methanol ...

OPTIMIZED HOST CELLS FOR THE PRODUCTION OF GLUTATHIONE

The disclosure concerns a genetically modified host cell for the production and accumulation of glutathione (GSH). The genetically modified host cell can allow the expression of a mutated Cys4p whose activity is increased. In addition or alternatively, the genetically modified host cell can express a mutated Yap1p whose translocation from the nucleus to the cytoplasm is reduced. Furthermore, in addition or alternatively, the genetically modified host cell can express an heterologous threonine aldolase (Gly1p). 1. A process of making glutathione , said process comprising fermenting a substrate with a genetically modified host cell to obtain a fermented mixture comprising glutathione , wherein the genetically modified host cell comprises at least one of:a first heterologous nucleic acid molecule coding for a mutated cystathionine beta-synthase protein (Cys4p) having an increased biological activity when compared to a wild-type Cys4p;a second heterologous nucleic acid molecule coding for a mutated Yap1p having a reduced ability of being translocated from nucleus to cytoplasm of the genetically modified host cell when compared to a wild-type Yap1p; anda third heterologous nucleic acid molecule coding for a threonine aldolase protein (Gly1p).2. The process of claim 1 , wherein the genetically modified host cell comprises the first heterologous nucleic acid molecule and at least one of the second nucleic acid molecule or the third heterologous nucleic acid molecule.3. The process of claim 1 , wherein the mutated Cys4p is a fragment of the wild-type Cys4p.4. The process of claim 3 , wherein the mutated Cys4p is obtained by deleting one or more C-terminal amino acid residue from the wild-type Cys4p.5. The process of claim 4 , wherein the mutated Cys4p is obtained by deleting a regulatory domain from the wild-type Cys4p.6. The process of claim 5 , wherein the mutated Cys4p consists of the amino acid sequence of SEQ ID NO: 2.7. The process of claim 1 , wherein claim 1 , in ...

ORGANIC ACID SYNTHESIS FROM C1 SUBSTRATES

Presented herein are biocatalysts and methods for converting C1-containing materials to organic acids such as muconic acid or adipic acid. 1. An engineered cell , comprising at least one exogenously added gene encoding a 3-dehydroshikimate dehydratase , a protocatechuic acid decarboxylase , a catechol 1 ,2-dioxygenase , aroG , trpE , a phosphoenolpyruvate synthase , or a transketolase; wherein the cell is able to convert a C1 substrate to an organic acid.2. The engineered cell of claim 1 , wherein the cell comprises genes encoding a 3-dehydroshikimate dehydratase claim 1 , a protocatechuic acid decarboxylase claim 1 , and a catechol 1 claim 1 ,2-dioxygenase.3Klebsiella variicola.. The engineered cell of claim 2 , wherein the 3-dehydroshikimate dehydratase is AroZ from4Enterobacter cloacae.. The engineered cell of claim 2 , wherein the protocatechuic acid decarboxylase is AroY from5Acinetobacter.. The engineered cell of claim 2 , wherein the catechol 1 claim 2 ,2-dioxygenase is CatA from6Klebsiella variicola,Enterobacter cloacae,Acinetobacter.. The engineered cell of claim 2 , wherein the 3-dehydroshikimate dehydratase is AroZ from the protocatechuic acid decarboxylase is AroY from and the catechol 1 claim 2 ,2-dioxygenase is CatA from7. The engineered cell of claim 1 , wherein the cell further comprises an exogenously added gene encoding a phosphoketolase.8. The engineered cell of claim 7 , wherein the phosphoketolase is PktA or PktB.9. The engineered cell of claim 1 , wherein the cell further comprises an exogenously added gene encoding a CbbL claim 1 , CbbS claim 1 , CbbQ or CbbP polypeptide.10. The engineered cell of claim 9 , wherein the cell further comprises exogenously added genes encoding CbbL claim 9 , CbbS claim 9 , CbbQ and CbbP polypeptides.11. The engineered cell of claim 1 , wherein the cell is a C1 metabolizing cell.12Methylomonas, Methylobacter, Methylococcus, Methylosinus, Methylocystis, Methylomicrobium, Methanomonas, Methylophilus, Methylobacillus ...

PHOSPHOKETOLASES FOR IMPROVED PRODUCTION OF ACETYL COENZYME A-DERIVED METABOLITES, ISOPRENE, ISOPRENOID PRECURSORS, AND ISOPRENOIDS

This present invention relates to cultured recombinant cells comprising a heterologous phosphoketolase (PKL) polypeptide that are capable of increased production of acetyl coenzyme A-derived metabolites, as well as methods for producing and using the same. In some embodiments, the recombinant cells further comprise one or more mevalonate (MVA) pathway polypeptides for the production of isoprenoid precursors, isoprene and isoprenoids. 1. A recombinant cell capable of increased carbon flux through the phosphoketolase pathway , wherein the recombinant cell comprises a heterologous nucleic acid sequence encoding a polypeptide having phosphoketolase activity , wherein the polypeptide comprises at least 65% sequence identity to SEQ ID NO:8.2. A recombinant cell capable of increased carbon flux through the phosphoketolase pathway , wherein the recombinant cell comprises: (i) a heterologous nucleic acid sequence encoding a polypeptide having phosphoketolase activity , wherein the polypeptide comprises at least 65% sequence identity to SEQ ID NO:8 and (ii) one or more nucleic acids encoding one or more polypeptides of the complete MVA pathway , wherein said recombinant cell comprising said polypeptide having phosphoketolase activity of (i) has a Performance Index value of greater than 1.0 in one or more of the following parameters: (a) cell growth on glucose , (b) cell growth on xylose , (c) production of intracellular acetyl-phosphate or (d) cell growth on glucose-6-phosphate.3. A recombinant cell capable of increased carbon flux through the phosphoketolase pathway , wherein the recombinant cell comprises: (i) a heterologous nucleic acid sequence encoding a polypeptide having phosphoketolase activity , wherein the polypeptide comprises at least 65% sequence identity to SEQ ID NO:8 and (ii) one or more nucleic acids encoding one or more polypeptides of the complete MVA pathway , wherein said polypeptide having phosphoketolase activity of (i) has a Performance Index value of ...

Engineering microbes and metabolic pathways for the production of ethylene glycol

The invention relates to recombinant cells and their use in the production of ethylene glycol.

GLUCONACETOBACTER HAVING ENHANCED CELLULOSE PRODUCTIVITY

A microorganism of the genus has enhanced cellulose productivity due to overexpression of fructose-bisphosphate aldolase, and optionally, phosphoglucomutase, UTP-glucose-1-phosphate uridylyltransferase, or cellulose synthase. A method of producing cellulose and a method of producing the microorganism are provided. 1Gluconacetobacter. A recombinant microorganism of the genus having enhanced cellulose productivity , the recombinant microorganism comprising a genetic modification that increases fructose-bisphosphate aldolase (FBA) activity.2. The microorganism of claim 1 , wherein the genetic modification is to increase the copy number of a gene encoding the fructose-bisphosphate aldolase.3. The microorganism of claim 2 , comprises an exogenous gene encoding the fructose-bisphosphate aldolase.4. The microorganism of claim 2 , wherein the fructose-bisphosphate aldolase belongs to EC 4.1.2.13.5. The microorganism of claim 1 , wherein the fructose-bisphosphate aldolase is a polypeptide having a sequence identity of 95% or more with respect to an amino acid sequence of SEQ ID NO: 1.6. The microorganism of claim 2 , wherein the gene has a nucleotide sequence of SEQ ID NO: 2.7Gluconacetobacter xylinus.. The microorganism of claim 1 , wherein the microorganism is8. The microorganism of claim 1 , further comprising one or more genetic modifications selected from the group consisting of a genetic modification that increases the activity of phosphoglucomutase (PGM) claim 1 , which catalyzes conversion of glucose-6-phosphate to glucose-1-phosphate; a genetic modification that increases the activity of UTP-glucose-1-phosphate uridylyltransferase (UPG) claim 1 , which catalyzes conversion of glucose-1-phosphate to UDP-glucose; and a genetic modification that increases the activity of cellulose synthase (CS) claim 1 , which catalyzes conversion of UDP-glucose to cellulose.9. The microorganism of claim 8 , wherein the microorganism has an increase in the copy number of one or more ...

Hydroxynitrile lyase

Provided are a method for obtaining an HNL gene and HNL derived from a millipede other than Chamberlinius hualienensis , and preparing a practically useable amount of HNL; and a method for producing optically active cyanohydrin using this HNL. A method for producing a millipede-derived HNL gene. A method that includes the selection of a gene having a base sequence that encodes a conserved amino acid sequence TAX1DIX2G (SEQ ID NO: 15) or VPNGDKIH (SEQ ID NO: 16) of millipede-derived HNL from genes present in an organism belonging to the Diplopoda. A protein having an amino acid sequence of any of (1)-(3) and having HNL activity. (1) An amino acid sequence listed in any of SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, 83, 85, 87, or 89; (2) an amino acid sequence having amino acids deleted, substituted, and/or added in an amino acid sequence of (1); or (3) an amino acid sequence having 90% or greater identity to an amino acid sequence of (1). A method for preparing optically active cyanohydrin by causing this millipede-derived HNL to act on a reaction solvent that contains an aldehyde or the like and a substance that generates hydrogen cyanide or the like.

METHODS FOR PRODUCING ISOPROPANOL AND ACETONE IN A MICROORGANISM

The present disclosure provides for novel metabolic pathways to increase acetone and isopropanol formation. More specifically, the present disclosure provides for a recombinant microorganism comprising a plurality of first native and/or heterologous enzymes that function in a first engineered metabolic pathway to convert fructose-6-phosphate to acetyl-CoA and acetate (e.g., phosphoketolase and acetate kinase), wherein the plurality of first native and/or heterologous enzymes is activated, upregulated, or overexpressed. The recombinant microorganism further comprises a plurality of second native and/or heterologous enzymes that function in a second engineered metabolic pathways to convert acetyl-CoA and acetate to isopropanol (e.g., thiolase, CoA transferase and acetoacetate decarboxylase), wherein the plurality of second native and/or heterologous enzymes is activated, upregulated, or overexpressed. Also provided are methods for making isopropanol or acetone using the recombinant microorganisms. 1. A recombinant microorganism comprising: a phosphoketolase; and', 'an acetate kinase; and, '(a) a plurality of first native and/or heterologous enzymes that function in a first engineered metabolic pathway to convert fructose-6-phosphate to acetyl-CoA and acetate, wherein the plurality of first native and/or heterologous enzymes is activated, upregulated, or overexpressed and comprises a thiolase;', 'a CoA transferase; and', 'an acetoacetate decarboxylase., '(b) a plurality of second native and/or heterologous enzymes that function in a second engineered metabolic pathways to convert acetyl-CoA and acetate to acetone and/or isopropanol, wherein the plurality of second native and/or heterologous enzymes is activated, upregulated, or overexpressed and comprises2. The recombinant microorganism of claim 1 , wherein the phosphoketolase:has the ability to convert D-xylulose 5-phosphate into D-glyceraldehyde 3-phosphate;has the ability to convert D-fructose 6-phosphate into D- ...

ORGANIC ACID SYNTHESIS FROM C1 SUBSTRATES

Presented herein are biocatalysts and methods for converting C1-containing materials to organic acids such as muconic acid or adipic acid. 1. An engineered cell , comprising an exogenously added gene encoding a 3-dehydroshikimate dehydratase (AroZ) , a protocatechuic acid decarboxylase (AroY) , a catechol 1 ,2-dioxygenase (CatA) , a phspho-2-dehydro-3-deoxyheptonate aldolase (AroG) , an anthranilate synthase (TrpE) , a phosphoenolpyruvate synthase , or a transketolase; wherein the engineered cell is able to convert a C1 substrate to an organic acid.2. The engineered cell of claim 1 , wherein the cell comprises genes encoding a 3-dehydroshikimate dehydratase (AroZ) claim 1 , a protocatechuic acid decarboxylase (AroY) claim 1 , and a catechol 1 claim 1 ,2-dioxygenase (CatA).3Klebsiella. The engineered cell of claim 2 , wherein the 3-dehydroshikimate dehydratase (AroZ) is from variicola.4Enterobacter cloacae.. The engineered cell of claim 2 , wherein the protocatechuic acid decarboxylase (AroY) is from5Acinetobacter.. The engineered cell of claim 2 , wherein the catechol 1 claim 2 ,2-dioxygenase (CatA) is from6KlebsiellaEnterobacter cloacaeAcinetobacter.. The engineered cell of claim 2 , wherein the 3-dehydroshikimate dehydratase (AroZ) is from variicola claim 2 , the protocatechuic acid decarboxylase (AroY) is from claim 2 , and the catechol 1 claim 2 ,2-dioxygenase (CatA) is from7. The engineered cell of claim 1 , wherein the cell further comprises an exogenously added gene encoding a phosphoketolase.8. The engineered cell of claim 7 , wherein the phosphoketolase is PktA or PktB.9. The engineered cell of claim 1 , wherein the cell further comprises an exogenously added gene encoding a CbbL claim 1 , CbbS claim 1 , CbbQ or CbbP polypeptide.10. The engineered cell of claim 9 , wherein the cell further comprises exogenously added genes encoding CbbL claim 9 , CbbS claim 9 , CbbQ and CbbP polypeptides.11. The engineered cell of claim 1 , wherein the engineered cell is a ...

Methanol dehydrogenase fusion proteins

Described herein are fusion proteins including methanol dehydrogenase (MeDH) and at least one other polypeptide such as 3-hexulose-6-phosphate dehydrogenase (HPS) or 6-phospho-3-hexuloisomerase (PHI), such as DHAS synthase or fructose-6-Phosphate aldolase or such as DHA synthase or DHA kinase. In a localized manner, the fusion protein can promote the conversion of methanol to formaldehyde and then to a ketose phosphate such as hexulose 6-phosphate or then to DHA and G3P. When expressed in cells, the fusion proteins can promote methanol uptake and rapid conversion to the ketose phosphate or to the DHA and D3P, which in turn can be used in a pathway for the production of a desired bioproduct. Beneficially, the rapid conversion to the ketose phosphate or to the DHA and G3P can avoid the undesirable accumulation of formaldehyde in the cell. Also described are engineered cells expressing the fusion protein, optionally include one or more additional metabolic pathway transgene(s), methanol metabolic pathway genes, target product pathway genes, cell culture compositions including the cells, methods for promoting production of the target product or intermediate thereof from the cells, compositions including the target product or intermediate, and products made from the target product or intermediate.

ALTERED HOST CELL PATHWAY FOR IMPROVED ETHANOL PRODUCTION

A recombinant yeast cell, fermentation compositions, and methods of use thereof are provided. The recombinant yeast cell includes at least one heterologous nucleic acid encoding one or more polypeptide having phosphoketolase activity; phosphotransacetylase activity; and/or acetylating acetaldehyde dehydrogenase activity, wherein the cell does not include a heterologous modified xylose reductase gene, and wherein the cell is capable of increased biochemical end product production in a fermentation process when compared to a parent yeast cell. 2. The recombinant yeast cell of claim 1 , wherein said cell has a reduced NAD-dependant glycerol phosphate dehydrogenase (GPD) activity when compared to a parent yeast cell.3. The recombinant yeast cell of claim 1 , wherein said cell comprises an altered pentose phosphate pathway resulting from one or more heterologously expressed nucleic acid affecting the pentose phosphate pathway.4. The recombinant yeast cell of claim 1 , wherein said biochemical end product is ethanol and it is produced at a level at least 0.5% higher to at least 15% higher than that produced in a parent yeast cell.5. (canceled)6. The recombinant cell of wherein:a) the phosphoketolase activity is encoded by a nucleic acid selected from at least one of the group consisting of a nucleic acid encoding SEQ ID NO: 56, SEQ ID NO: 54, SEQ ID NO: 48, SEQ ID NO: 3, SEQ ID NO: 44, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO:66, SEQ ID NO:72, and a nucleic acid having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 56, SEQ ID NO: 54, SEQ ID NO: 48, SEQ ID NO: 3, SEQ ID NO: 44, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO:66 or SEQ ID NO:72;b) the phophotransacetylase activity is encoded by a nucleic acid comprising SEQ ID NO: 4 or having at least 80%, 85%, 90%, 95%, 98% or 99% identity to SEQ ID NO: 4; and/orc) the acetylating acetaldehyde dehydrogenase activity is encoded by a nucleic acid selected from at least one of the group ...

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.

CATALYTICALLY ACTIVE PROTEIN AGGREGATES AND METHODS FOR PRODUCING THE SAME

Disclosed are catalytically active water-insoluble protein aggregates comprising fusion proteins which comprise a coiled-coil domain and a catalytic domain, methods of manufacturing such protein aggregates, and their use. 1. A water-insoluble protein aggregate comprising a fusion protein , wherein said fusion protein comprises a coiled-coil domain and a catalytic domain.2. The protein-aggregate according to claim 1 , wherein the fusion protein is a N-terminal fusion protein.3. The protein aggregate according to claim 1 , wherein the fusion protein is a C-terminal fusion protein.4. The protein aggregate according to claim 1 , wherein the coiled-coil domain is selected from the group consisting ofdomains comprising at least three heptad repeats, the amino acid sequence of said heptad repeats being represented by hxxhcxc, wherein h represents a hydrophobic amino acid residue, c represents a charged amino acid residue, and x represents any amino acid residue;domains wherein the PCOILS webserver predicts the heptad repeat pattern hxxhcxc, wherein h represents a hydrophobic amino acid residue, c represents a charged amino acid residue, and x represents any amino acid residue, in any of the 3 sequence windows (14, 21, 28 amino acids) with a score above 0.2 over 3 heptad repeats, using the MTIDK matrix;domains wherein the MARCOILS webserver predicts at least 1 coiled-coil domain with a threshold of 1.0 within the sequence, using the 9FAM matrix; anddomains comprising a repeat pattern comprising two or more hydrophobic amino acid residues and two or more charged amino acid residues within a stretch of seven amino acid residues, wherein the PCOILS webserver predicts the repeat pattern comprising two or more hydrophobic amino acid residues and two or more charged amino acid residues within a stretch of seven amino acid residues with a score above 0.2 over two repeats in any of the three sequence windows (14, 21, 28 amino acids).5. The protein aggregate according to claim 1 , ...

A recombinant strain having modified sugar metabolic pathway and method for screening sugar isomerase using same

The present invention relates to a recombinant strain having a modified sugar metabolic pathway resulting from the introduction of an enzyme derived from a different strain to a strain, and a high-speed screening method for various mutants and variants by which useful materials can be obtained or which can produce useful materials. The use of a recombinant vector and a strain according to the present invention not only can construct a new metabolic pathway in a strain to effectively obtain D-tagatose from D-galactose, but also can introduce randomly modified sugar isomerases and then allow D-galactose isomerase variants to be rapidly screened by conducting a cell growth-associated screening system.

MICROBES & METHODS FOR IMPROVED CONVERSION OF A FEEDSTOCK

Genetically engineered cells and methods are presented that enhance the consumption of xylose in a medium comprising a mix of five- and six-carbon sugars. Method of using these microbes to enhance xylose utilization and methods of making value products using these microbes are also disclosed herein. 1. A microbial cell comprising (1) a non-native xylose isomerase (xylA) gene; and (2) a non-native fructose-6-phosphate phosphoketolase (xfp) gene , and/or (3) a d-xylose ABC transporter (xylFGH).2. The microbial cell of claim 1 , wherein the microbial cell further comprises: (4) a non-native xylulose kinase (xylB) gene.3. The microbial cell of claim 1 , wherein the microbial cell is a eukaryote or a prokaryote.4Saccharomyces, Pichia, CandidaAspergillus.. The microbial cell of claim 3 , wherein the eukaryote belongs to a genus selected from the group consisting of claim 3 , and5Escherichia, Bacillus, Corynebacterium, Alcaligenus, Zymomonas, Clostridium, Lactobacillus, SynechococcusSynechocystis.. The microbial cell of claim 3 , wherein the prokaryote belongs to a genus selected from the group consisting of claim 3 , and6. The microbial cell of claim 1 , wherein the xylA gene comprises a coding sequence that is a native sequence claim 1 , located on the chromosome claim 1 , and the promoter driving the xylA is heterologous to the xylA coding sequence.7. The microbial cell of claim 1 , wherein the xfp gene comprises a nucleotide sequence that encodes a protein with at least 85% sequence identity to SEQ ID NO:1 or SEQ ID NO:2.8. The microbial cell of claim 7 , wherein the encoding nucleotide sequence is SEQ ID NO:3 or SEQ ID NO:4.9. The microbial cell of claim 1 , wherein the xylFGH gene comprises a nucleotide sequence that encodes a protein with at least 85% sequence identity to SEQ ID NO:5.10. A method of improving xylose utilization in a microbial cell expressing xylose isomerase (XylA) and xylulose kinase (XylB) genes claim 1 , the method comprising genetically ...

CYB5 AND CYP17 MUTATIONS FOR ALTERATION OF 16-ANDROSTENE STEROID SYNTHESIS AND REDUCED BOAR TAINT IN PIGS

Novel mutations in cytochrome P450C17 (CYP17) and cytochrome b5 (CYB5) affecting 16-androstene steroid synthesis are disclosed. The novel mutations result in alterations in production of critical intermediaries in the synthesis of 16-androstene steroids. Altering the activity of these enzymes may be useful in enhancing reducing androstenone synthesis and reducing boar taint. The identification of these novel mutations also allows for the development of transgenic pigs bearing mutations in these enzymes or for genetic screening to identify pigs on the basis of their CYP17 and/or CYB5 genotype. Pigs having these mutations may be selected and bred to produce pigs that have a lower incidence of boar taint. 1. An altered CYP17 protein that modifies 16-androstene steroid activity or production in pigs , said protein comprising:a modification of the amino acid present at one or more positions selected from the group consisting of: amino acids 102, 103, 104, 106, 108, 109, 112, 202, 344, 345, 348, 352 and 454 of the porcine CYP17 protein, compared to wild type (SEQ ID NO:4).2. The altered CYP17 protein of wherein said modification comprises one of more of:(a) a glutamine residue at position 102;(b) a serine residue position 103;(c) a leucine residue at position 104;(d) an alanine residue at position 106;(e) an aspartic acid residue at position 106;(f) a glutamine residue at position 108;(g) a glycine residue at position 109;(h) a valine residue at position 112;(i) a threonine residue at position 202;(j) a phenylalanine residue at position 344;(k) an asparagine residue at position 345;(l) a serine residue at position 348;(m) a methionine residue at position 352; and(n) a valine residue at position 454.3. A method of modifying 16-androstene steroid activity to reducing boar taint in a pig comprising:introducing to said pig a polynucleotide that encodes a CYP17 protein, wherein amino acid 102, 103, 104, 106, 108, 112, 202, 344, 345, 348, 352 and 454 of the porcine CYP17 ...

ENZYMATIC PRODUCTION OF ACETYL PHOSPHATE FROM FORMALDEHYDE

Described is a method for the enzymatic production of acetyl phosphate from formaldehyde using a phosphoketolase or a sulfoacetaldehyde acetyltransferase. 1. A method for the enzymatic production of acetyl phosphate from formaldehyde and phosphate in which the conversion of acetyl phosphate from formaldehyde and phosphate is achieved by the use of a phosphoketolase or of a sulfoacetaldehyde acetyltransferase (EC 2.3.3.15) according to the following reaction scheme:{'br': None, 'sub': 2', '2, '2CHO+phosphate→acetyl phosphate+HO.'}2. The method of claim 1 , wherein the phosphoketolase is(a) a phosphoketolase (EC 4.1.2.9), or(b) a fructose-6-phosphate phosphoketolase (EC 4.1.2.22).3. The method of which further comprises the step of converting the produced acetyl phosphate into acetate.4. The method of claim 3 , wherein the conversion of acetyl phosphate into acetate is achieved by making use of an acetate kinase (EC 2.7.2.1) or of a butyrate kinase (EC 2.7.2.7) or of an acetate kinase (diphosphate) (EC 2.7.2.12) or of a propionate kinase (EC 2.7.2.15) or of an acylphosphatase (EC 3.6.1.7).5. The method of which further comprises the step of enzymatically converting the produced acetyl phosphate into acetyl-coenzyme A.6. The method of claim 5 , wherein the conversion of acetyl phosphate into acetyl-coenzyme A is achieved by making use of a phosphate acetyltransferase (EC 2.3.1.8).7. The method of further comprising the step of providing the formaldehyde to be converted into acetyl phosphate by enzymatically converting methanol into formaldehyde.8. The method of wherein the enzymatic conversion of methanol into formaldehyde is achieved by making use of a methanol dehydrogenase (EC 1.1.1.244) or a methanol dehydrogenase (cytochrome c) (EC 1.1.2.7) or an alcohol oxidase (EC 1.1.3.13).9. A composition containing(a) formaldehyde and a phosphoketolase and/or a sulfoacetaldehyde acetyltransferase; or(b) formaldehyde and a microorganism expressing a phosphoketolase and/or a ...

Recombinant yeast and method for producing ethanol using the same

This invention is aimed at improving an ethanol fermentation ability of a recombinant yeast strain having an ability of assimilating pentose, such as xylose or arabinose. The recombinant yeast strain haying an ability of assimilating pentose is obtained by lowering activity of a gene involved in upstream of glyceraldehyde-3-phosphate in the Embden-Meyerhof pathway.

RECOMBINANT MICROORGANISM METABOLIZING 3,6-ANHYDRIDE- L-GALACTOSE AND A USE THEREOF

The present invention relates to a recombinant microorganism metabolizing 3,6-anhydro-L-galactose and a use thereof, and, more particularly, can produce ethanol from a recombinant microorganism expressing an enzyme group involved in a metabolic pathway of 3,6-AHG. 1. A recombinant vector for producing ethanol , comprising:a gene encoding 3,6-anhydro-L-galactose dehydrogenase;a gene encoding 3,6-anhydrogalactonic acid cycloisomerase;a gene encoding 2-keto-3-deoxy-galactonic acid kinase; anda gene encoding 2-keto-3-deoxy-phosphogalactonic acid aldolase.2. The recombinant vector of claim 1 , wherein the gene encoding 3 claim 1 ,6-anhydro-L-galactose dehydrogenase is represented by the base sequence set forth in SEQ ID NO: 1.37. The recombinant vector of claim 1 , wherein the gene encoding 3 claim 1 ,6-anhydrogalactonic acid cycloisomerase is represented by any one of the base sequences set forth in SEQ ID NOs: 3 claim 1 , 5 and .4. The recombinant vector of claim 1 , wherein the gene encoding 2-keto-3-deoxy-galactonic acid kinase is represented by the base sequence set forth in SEQ ID NO: 9.5. The recombinant vector of claim 1 , wherein the gene encoding 2-keto-3-deoxy-phosphogalactonic acid aldolase is represented by the base sequence set forth in SEQ ID NO: 11.6. A recombinant microorganism for producing ethanol claim 1 , which is transformed by:a gene encoding 3,6-anhydro-L-galactose dehydrogenase;a gene encoding 3,6-anhydrogalactonic acid cycloisomerase;a gene encoding 2-keto-3-deoxy-galactonic acid kinase; anda gene encoding 2-keto-3-deoxy-phosphogalactonic acid aldolase.7. The recombinant microorganism of claim 6 , wherein the recombinant microorganism is transformed with:a recombinant vector comprising a gene encoding 3,6-anhydro-L-galactose dehydrogenase;a recombinant vector comprising a gene encoding 3,6-anhydrogalactonic acid cycloisomerase;a recombinant vector comprising a gene encoding 2-keto-3-deoxy-galactonic acid kinase; anda recombinant vector ...

ENGINEERED MICROORGANISMS WITH G3P -> 3PG ENZYME AND/OR FRUCTOSE-1,6-BISPHOSPHATASE INCLUDING THOSE HAVING SYNTHETIC OR ENHANCED METHYLOTROPHY

Described herein are engineered cells including ones having synthetic methylotrophy which include an NADH-dependent enzyme capable of converting G3P to 3PG (e.g., gapN) and/or fructose-1,6-bisphosphatase, along with hexulose-6-phosphate synthase, 6-phospho-3-hexuloisomerase, a phosphoketolase, or a combination thereof. Engineered cells of the disclosure beneficially maintain adequate pool sizes of phosphorylated C3 and/or C4 compounds, and/or provide increased levels of NADPH. As such, the modifications allow for the generation of C6 compounds from C1 (e.g., a methanol feedstod) and C5 compounds, the regeneration of C5 compounds from C6 compounds by carbon rearrangement, and an improved balance between regeneration of C5 compounds and lower glycolysis. In turn, this allows the engineered microorganism to generate sufficient quantities of metabolic precursors (e.g., acetyl-CoA) which can be used in a bioproduct pathway, and the engineered cells can include further modifications to those pathway enzymes allowing for production of a desired bioproduct. 1. An engineered microorganism having synthetic or enhanced methylotrophy comprising:{'i': 'B. methanolicus', '(a) exogenous enzyme A that (ai) is capable of converting glyceraldehyde 3-phosphate (G3P) to 3-phosphoglycerate (3PG), (aii) has at least 50% sequence identity to SEQ ID NO:1 (gapN), wherein (ai) or (aii) is capable of reducing NADP to NADPH, or (aiii) that is a fructose-1,6-bisphosphatase; and'}(b) an exogenous enzyme B which is (bi) a phosphoketolase, (bii) a hexulose-6-phosphate synthase, (biii) 6-phospho-3-hexuloisomerase, or any combination of (bi), (bii) and (biii).2. The engineered microorganism of comprising the (a) exogenous enzyme A claim 1 , and the (bi) exogenous phosphoketolase claim 1 , and optionally the (bii) exogenous hexulose-6-phosphate synthase claim 1 , and the (biii) exogenous 6-phospho-3-hexuloisomerase.3. (canceled)4. (canceled)5B. methanolicus. An engineered microorganism comprising an ...

PHOSPHOKETOLASES FOR IMPROVED PRODUCTION OF ACETYL COENZYME A-DERIVED METABOLITES, ISOPRENE, ISOPRENOID PRECURSORS, AND ISOPRENOID

This present invention relates to cultured recombinant cells comprising heterologous phosphoketolase (PKL) polypeptides that are capable of increased production of acetyl coenzyme A-derived metabolites, as well as methods for producing and using the same. In some embodiments, the recombinant cells further comprise one or more mevalonate (MVA) pathway polypeptides for the production of isoprenoid precursors, isoprene and isoprenoids. 161.-. (canceled)62. A recombinant cell capable of increased carbon flux through the phosphoketolase pathway , wherein the recombinant cell comprises:(a) a heterologous nucleic acid sequence encoding a polypeptide having phosphoketolase activity, wherein the polypeptide comprises at least 65% sequence identity to SEQ ID NO: 8 or SEQ ID NO: 11;(b) attenuated activity of an endogenous acetate kinase enzyme; and(c) one or more nucleic acids encoding one or more polypeptides of the complete mevalonate (MVA) pathway,{'i': 'E. gallinarum', '(1) wherein said recombinant cell comprising said polypeptide having phosphoketolase activity of (i) has a Performance Index value of greater than 1.0 in one or more of the following parameters: (a) cell growth on glucose, (b) cell growth on xylose, (c) production of intracellular acetyl-phosphate, or (d) cell growth on glucose-6-phosphate, wherein the Performance Index value is calculated as activity per unit relative to a corresponding cell expressing a phosphoketolase from ; or'}{'i': 'E. gallinarum.', '(2) wherein said polypeptide having phosphoketolase activity of (i) has a Performance Index value of greater than 1.0 in one or more of the following parameters: (e) protein solubility, (f) protein expression, or (g) fructose-6-phosphate (F6P) Specific Activity, wherein the Performance Index value is calculated as activity per unit relative to a phosphoketolase from'}63. The recombinant cell of claim 62 , wherein the polypeptide comprises at least 90% sequence identity to a polypeptide selected from the ...

Modulation of sphingosine 1-phosphate metabolizing enzymes for the treatment of negative-strand rna virus infections

The present invention relates to compounds and methods for the prevention or treatment of infections by negative strand RNA viruses, such as influenza virus and measles virus, wherein said compounds delay or inhibit viral replication by modulating the level or activity of a polypeptide involved in the synthesis or degradation of sphingosine-1-phosphate (S1P) in a cell, tissue, or subject. The methods involve administration of one or more compounds which modulate the level of gene expression, where the gene encodes a polypeptide involved in regulating the metabolic level of S1P, or modulate the level or activity of a polypeptide involved in regulating the metabolic level of S1P, such as sphingosine kinase (SK) and S1P lyase (SPL). Exemplary methods are directed towards reducing the level of SW by reducing the level or activity of one or more SKs, increasing the level or activity of one or more SPLs, or a combination of both steps.

Microbial production of renewable glycolate

Some aspects provide engineered microbes for glycolate production. Methods for microbe engineering and culturing are also provided herein. Such engineered microbes exhibit greatly enhanced capabilities for glycolate production.

Byosynthetic Production of Acyl Amino Acids

The present invention relates to a cell for producing acyl glycinates wherein the cell is genetically modified to comprise 115-. (canceled)16. A cell for producing acyl glycinates , wherein said cell is genetically modified to comprise:a) at least a first genetic mutation that, relative to the wild type cell, increases the expression of an amino acid-N-acyl-transferase;b) at least a second genetic mutation that, relative to the wild type cell, increases the expression of an acyl-CoA synthetase; andc) at least a third genetic mutation that, relative to the wild type cell, decreases the expression of at least one enzyme selected from the group consisting of: an enzyme of the glycine cleavage system; glycine hydroxymethyltransferase (GlyA); threonine aldolase (LtaE); threonine dehydrogenase (Tdh); 2-Amino-3-Ketobutyrate CoA-Ligase (Kbl); and allothreonine dehydrogenase (YdfG).17. The cell of claim 16 , wherein said cell comprises a mutation in an enzyme from the glycine cleavage system selected from the group consisting of: glycine cleavage system T protein; glycine cleavage system H protein; and glycine cleavage system P protein.18. The cell of claim 16 , wherein the third genetic mutation decreases the expression relative to the wild type cell of the glycine hydroxymethyltransferase (GlyA) claim 16 , the threonine aldolase (LtaE) claim 16 , the glycine cleavage system T protein claim 16 , the glycine cleavage system H protein and the glycine cleavage system P protein.19. The cell of claim 18 , wherein the glycine cleavage system T protein has 85% sequence identity to SEQ ID NO:58 claim 18 , the glycine cleavage system H protein has 85% sequence identity to SEQ ID NO:59 claim 18 , and the glycine cleavage system P protein has 85% sequence identity to SEQ ID NO:60.20. The cell of claim 16 , wherein the glycine hydroxymethyltransferase (GlyA) has 85% sequence identity to SEQ ID NO:61 and the threonine aldolase (LtaE) has 85% sequence identity to SEQ ID NO:62.21. The ...

ALTERED HOST CELL PATHWAY FOR IMPROVED ETHANOL PRODUCTION

A recombinant yeast cell, fermentation compositions, and methods of use thereof are provided. The recombinant yeast cell includes at least one heterologous nucleic acid encoding one or more polypeptide having phosphoketolase activity; phosphotransacetylase activity; and/or acetylating acetaldehyde dehydrogenase activity, wherein the cell does not include a heterologous modified xylose reductase gene, and wherein the cell is capable of increased biochemical end product production in a fermentation process when compared to a parent yeast cell. 1. A recombinant yeast cell comprising at least one heterologous nucleic acid encoding one or more polypeptide having:i) phosphoketolase activity;ii) phosphotransacetylase activity; and wherein said cell does not comprise a heterologous modified xylose reductase gene,', 'wherein said cell is capable of increased ethanol production from glucose in a fermentation process when compared to the yeast cell without the at least one heterologous nucleic acid, and', {'i': 'Lactobacillus plantarum.', 'wherein the polypeptide having phosphoketolase activity has the amino acid of SEQ ID NO: 57, the polypeptide having acetylating acetaldehyde dehydrogenase activity has the amino acid of SEQ ID NO: 32, and the polypeptide having phophotransacetylase activity is the phophotransacetylase from'}], 'iii) acetylating acetaldehyde dehydrogenase activity,'}2. The recombinant yeast cell of claim 1 , wherein said cell has a reduced NAD-dependant glycerol phosphate dehydrogenase (GPD) activity when compared to a parent yeast cell.3. The recombinant yeast cell of claim 1 , wherein said cell comprises an altered pentose phosphate pathway resulting from one or more heterologously expressed nucleic acid affecting the pentose phosphate pathway.4Saccharomyces cerevisiae.. The recombinant yeast cell of wherein the species of the yeast cell is5. The recombinant yeast cell of claim 1 , wherein said fermentation process is selected from the group consisting of ...

ENGINEERED ALDOLASE POLYPEPTIDES AND USES THEREOF

The present invention provides engineered polypeptides that are useful for the asymmetric synthesis of β-hydroxy-α-amino acids under industrial-relevant conditions. The present disclosure also provides polynucleotides encoding engineered polypeptides, host cells capable of expressing engineered polypeptides, and methods of producing β-hydroxy-α-amino acids using engineered polypeptides. Compared to other processes of preparation, the use of the engineered polypeptides of the present invention for the preparation of β-hydroxy-α-amino acids results in high purity of the desired stereoisomers, mild reaction conditions, low pollution and low energy consumption. So, it has good industrial application prospects. 1. An engineered aldolase polypeptide comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2 or SEQ ID NO: 4 that is , under suitable reaction conditions , capable of condensing 4-(methylsulfonyl)benzaldehyde with glycine to produce (2S ,3R)-2-amino-3-hydroxy-3-[4-(methylsulfonyl)phenyl] propanoic acid in a diastereomeric excess of at least 60%.2. The aldolase polypeptide of claim 1 , wherein said suitable reaction conditions include about 30 g/L 4-(methylsulfonyl)benzaldehyde claim 1 , about 123 g/L glycine claim 1 , about 50 μM pyridoxal 5′-phosphate (PLP) claim 1 , and about 20% (v/v) DMSO claim 1 , at about 30° C.3. The aldolase polypeptide of claim 1 , wherein the amino acid sequence of the aldolase polypeptide comprises an amino acid sequence that differs from the sequence of SEQ ID NO: 4 at one or more amino acid residues selected from among: 16 claim 1 , 18 claim 1 , 19 claim 1 , 37 claim 1 , 38 claim 1 , 41 claim 1 , 42 claim 1 , 43 claim 1 , 57 claim 1 , 61 claim 1 , 86 claim 1 , 106 claim 1 , 110 claim 1 , 130 claim 1 , 151 claim 1 , 152 claim 1 , 159 claim 1 , 198 claim 1 , 201 claim 1 , 249 claim 1 , 256 claim 1 , 263 claim 1 , 266 claim 1 , 310 claim 1 , 327 claim 1 , 335 claim 1 , 337 claim 1 , 338 claim 1 , 339 ...

Microorganism of the genus escherichia producing l-tryptophan and method for producing l-tryptophan using the same

The present application relates to a microorganism of the genus Escherichia producing L-tryptophan and, more specifically, to a microorganism of the genus Escherichia with improved activity of producing L-tryptophan by weakening or inactivating the activity of endogenous 6-phosphogluconate dehydratase and 2-keto-3-deoxy-6-phosphogluconate aldolase. Additionally, the present application relates to a method for producing L-tryptophan using the microorganism of the genus Escherichia.

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

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

Epitopes

The present invention relates to epitopes containing homocitrulline (Hcit) that can be used as targets for cancer immunotherapy. The homocitrullinated T cell epitope has (i) a predicted binding score to MHC class II or class I of <30 using the online IEDB prediction program (http://www.iedb.org/) and (ii) at least 5 consecutive amino acids that form a spiral conformational structure. These modified peptides can be used as vaccines or as targets for T cell receptor (TCR) and adoptive T cell transfer therapies.

RECOMBINANT MICROORGANISM FOR THE PRODUCTION OF USEFUL METABOLITES

Described are recombinant microorganisms characterized by having phosphoketolase activity, having a diminished or inactivated Embden-Meyerhof-Parnas pathway (EMPP) by inactivation of the gene(s) encoding phosphofructokinase or by reducing phosphofructokinase activity as compared to a non-modified microorganism and having a diminished or inactivated oxidative branch of the pentose phosphate pathway (PPP) by inactivation of the gene(s) encoding glucose-6-phosphate dehydrogenase or by reducing glucose-6-phosphate dehydrogenase activity as compared to a non-modified microorganism. These microorganisms can be used for the production of useful metabolites such as acetone, isobutene or propene. 116-. (canceled)17. A genetically modified prokaryotic microorganism comprising the following characteristics:(A) increased phosphoketolase activity as compared to a non-genetically modified microorganism; and '(2) not possessing phosphofructokinase activity; and', '(B) (1) diminished or inactivated phosphofructokinase activity as compared to a non-genetically modified microorganism; or'} '(2) not possessing glucose-6-phosphate dehydrogenase activity.', '(C) (1) diminished or inactivated glucose-6-phosphate dehydrogenase activity as compared to a non-genetically modified microorganism; or'}18E. coli. The genetically modified prokaryotic microorganism of claim 17 , wherein the genetically modified microorganism is comprising the genotype Azwf_edd_eda::FRT ΔpfkA::FRT ΔpfkB::aad+.19E. coli.. The genetically modified prokaryotic microorganism of claim 17 , wherein the genetically modified microorganism is20. The genetically modified prokaryotic microorganism of claim 17 , wherein the non-genetically modified microorganism does not have phosphoketolase activity.21. The genetically modified prokaryotic microorganism of claim 20 , wherein the non-genetically modified microorganism is genetically modified so as to comprise a nucleotide sequence encoding a phosphoketolase.22. The genetically ...

MICROORGANISM HAVING ENHANCED CELLULOSE PRODUCTIVITY, METHOD OF PRODUCING CELLULOSE BY USING THE SAME, AND METHOD OF PRODUCING THE MICROORGANISM

Provided are a microorganism having enhanced cellulose productivity, a method of producing cellulose by using the microorganism, and a method of producing the microorganism. 1. A recombinant microorganism comprising a genetic modification that enhances expression of at least one gene regulated by a glycerol operon selected from a gene encoding glycerol-3-phosphate dehydrogenase (glpD) , a gene encoding glycerol kinase (glpK) , a gene encoding fructose-1 ,6-bisphosphatase (glpX) , and a gene encoding fructose-bisphosphate aldolase (FBA) 3.2. The recombinant microorganism of claim 1 , wherein the genetic modification comprises at least one modification selected from (i) a disruptive mutation of a regulatory element of the glycerol operon and (ii) substitution of an operator binding site or native promoter with a constitutive promoter.3. The recombinant microorganism of claim 1 , wherein the genetic modification is attenuation or inactivation of a glycerol-3-phosphate repressor.4. The recombinant microorganism of claim 2 , wherein the genetic modification is substitution of a promoter of the glycerol operon with a constitutive promoter.5. The recombinant microorganism of claim 5 , wherein the constitutive promoter is a tac promoter or a gap promoter.6. The recombinant microorganism of claim 1 , wherein the genetic modification increases the expression of the gene encoding fructose-1 claim 1 ,6-bisphosphatase (glpX) and the gene encoding fructose-bisphosphate aldolase (FBA) 3.7. The recombinant microorganism of claim 1 , wherein the genetic modification increases a copy number of the at least one gene.8. The recombinant microorganism of claim 1 , wherein the glycerol-3-phosphate dehydrogenase (glpD) belongs to EC 1.1.5.3 claim 1 , EC 1.1.1.94 claim 1 , or EC 1.1.1.8 claim 1 , the glycerol kinase (glpK) belongs to EC 2.7.1.30 claim 1 , the fructose-1 claim 1 ,6-bisphosphatase (glpX) belongs to EC 3.1.3.11 claim 1 , and the fructose-bisphosphate aldolase (FBA) 3 belongs ...

CELL-FREE SYSTEM FOR CONVERTING METHANE INTO FUEL AND CHEMICAL COMPOUNDS

The present disclosure relates, in some aspects, to cell-free methods and systems for large-scale conversion of methane to isobutanol, comprising combining, in a bioreactor at elevated pressure, methane, oxygen, and cell lysates containing methane monooxygenase, methanol dehydrogenase, and enzymes that catalyze the conversion of formaldehyde to isobutanol, to form a cell-free reaction mixture, and incubating under suitable conditions the cell-free reaction to convert methane to isobutanol. 175-. (canceled)76. A composition comprising at least two cell lysates and the following enzymes: a methane monooxygenase , a methanol dehydrogenase , a hexulose-6-phosphate synthase , a 6-phospho-3-hexuloisomerase , a 6-phosphofructokinase , a fructose bisphosphate aldolase , a triose phosphate isomerase , a transketolase , a ribose-5-phosphate isomerase or a ribulose-5-phosphate 3-epimerase , a glyceraldehyde 3-phosphate dehydrogenase , a phosphoglycerate kinase , a phosphoglycerate mutase , an enolase , and a pyruvate kinase , wherein at least one of the cell lysates is obtained from recombinant cells that overexpress at least one of the foregoing enzymes.77. The composition of claim 76 , wherein(i) the methane monooxygenase has EC number 1.14.13.25 or 1.14.18.3,(ii) the methanol dehydrogenase has EC number 1.1.1.244, 1.1.2.7, or 1.1.99.37,(iii) the hexulose-6-phosphate synthase has EC number 4.1.2.43,(iv) the 6-phospho-3-hexuloisomerase has EC number 5.3.1.27,(v) the 6-phosphofructokinase has EC number 2.7.1.11,(vi) the fructose bisphosphate aldolase has EC number 4.1.2.13,(vii) the triose phosphate isomerase has EC number 5.3.1.1,(viii) the transketolase has EC number 2.2.1.1,(xi) the ribulose-5-phosphate 3-isomerase has EC number 5.3.1.6, or the ribulose-5-phosphate 3-epimerase has EC number 5.1.3.1,(xii) the glyceraldehyde 3-phosphate dehydrogenase has EC number 1.2.1.12,(xiii) the phosphoglycerate kinase has EC number 2.7.2.3,(xiv) the phosphoglycerate mutase has EC number ...

Metabolic Pathways with Increased Carbon Yield

The present invention relates to the conversion of a carbon source into acetyl phosphate with increased carbon yield. In particular, the invention provides metabolically engineered micro-organisms capable of producing acetyl phosphate from a carbon source with increased carbon yield, which micro-organisms have been transformed with at least one exogenous nucleic acid encoding a phosphoketolase having sedoheptulose-7-phosphate phosphoketolase activity and which are further genetically modified to have eliminated transketolase activity. The invention also provides methods for the production of chemicals using said micro-organisms. 115.-. (canceled)16. A metabolically engineered micro-organism capable of increased acetyl phosphate conversion from a carbon source compared to a corresponding non-metabolically engineered micro-organism , wherein the micro-organism comprises at least one exogenous nucleic acid encoding a phosphoketolase having sedoheptulose-7-phosphate phosphoketolase activity , and wherein the micro-organism is further genetically modified to have eliminated transketolase activity.17Bifidobacterium longum. The micro-organism according to claim 16 , wherein the phosphoketolase having sedoheptulose-7-phosphate phosphoketolase activity is a phosphoketolase or a functional variant thereof claim 16 , preferably the phosphoketolase comprising a sequence set forth in SEQ ID NO:2.18. The micro-organism according to claim 16 , wherein the micro-organism is further genetically modified to facilitate a metabolic pathway that converts a fructose-6-phosphate molecule and 2 phosphate molecules into 3 molecules acetyl-phosphate and 2 molecules water.19. The micro-organism according to claim 18 , wherein the micro-organism is genetically modified to have eliminated transaldolase activity.20. The micro-organism according to claim 19 , wherein the micro-organism is genetically modified to express or overexpress at least one enzyme selected from the group comprising an ...

Recombinant microorganism for the production of useful metabolites

Described are recombinant microorganisms characterized by having phosphoketolase activity, having a diminished or inactivated Embden-Meyerhof-Parnas pathway (EMPP) by inactivation of the gene(s) encoding phosphofructokinase or by reducing phosphofructokinase activity as compared to a non-modified microorganism and having a diminished or inactivated oxidative branch of the pentose phosphate pathway (PPP) by inactivation of the gene(s) encoding glucose-6-phosphate dehydrogenase or by reducing glucose-6-phosphate dehydrogenase activity as compared to a non-modified microorganism. These microorganisms can be used for the production of useful metabolites such as acetone, isobutene or propene.

NOVEL (R)-HYDROXYNITRILE LYASE

The objective of the present invention is to provide a (R)-hydroxynitrile lyase which is more stable than a hydroxynitrile lyase derived from a plant, a gene which encodes the hydroxynitrile lyase and by which heterologous expression is possible, and a method for producing the hydroxynitrile lyase. The (R)-hydroxynitrile lyase according to the present invention is characterized in having the specific amino acid sequence such as the amino acid sequence of SEQ ID NO: 3. 1. A (R)-hydroxynitrile lyase having any one of the following amino acid sequences (1) to (3):(1) an amino acid sequence of SEQ ID NO: 3;(2) an amino acid sequence specified in the (1) with deletion, substitution and/or addition of 1 or more amino acid residues, wherein a hydroxynitrile lyase activity of a (R)-hydroxynitrile lyase having the amino acid sequence (2) is similar to or superior to that of natural (R)-hydroxynitrile lyase having the amino acid sequence of SEQ ID NO: 3;(3) an amino acid sequence having at least 30% sequence homology to the amino acid sequence specified in the (1), wherein a hydroxynitrile lyase activity of a (R)-hydroxynitrile lyase having the amino acid sequence (3) is similar to or superior to that of natural (R)-hydroxynitrile lyase having the amino acid sequence of SEQ ID NO: 3.2. The (R)-hydroxynitrile lyase according to claim 1 , being a dimer of a subunit having any one of the amino acid sequences (1) to (3).3Chamberlinius.. The (R)-hydroxynitrile lyase according to claim 1 , derived from genus4Chamberlinius hualienensis.. The (R)-hydroxynitrile lyase according to claim 3 , derived from5. A (R)-hydroxynitrile lyase gene having any one of the following base sequences (4) to (6):(4) a base sequence of SEQ ID NO: 2;(5) a base sequence specified in the (4) with deletion, substitution and/or addition of 1 or more bases, wherein a hydroxynitrile lyase activity of a (R)-hydroxynitrile lyase encoded by the gene having the base sequence (5) is similar to or superior to that of ...

RECOMBINANT MICROORGANISMS HAVING A METHANOL ELONGATION CYCLE (MEC)

Provided are microorganisms that catalyze the synthesis of chemicals and biochemicals from a methanol, methane and/or formaldehyde. Also provided are methods of generating such organisms and methods of synthesizing chemicals and biochemicals using such organisms. 1. A recombinant prokaryote or yeast microorganism comprising a metabolic pathway for the synthesis of acetyl phosphate from methanol , methane or formaldehyde using a pathway comprising an enzyme having fructose-6-phosphoketolase (Fpk) activity and/or xylulose-5-phosphoketolase (Xpk) activity with an acetyl-phosphate yield better than a wild-type or parental organism lacking Fpk and/or Xpk.23-. (canceled)4E. coli. The recombinant microorganism of claim 1 , wherein the microorganism is derived from an microorganism.56-. (canceled)7. The recombinant microorganism of any of claim 1 , wherein the microorganism is engineered to heterologously expresses one or more of the following enzymes:(a) a phosphoketolase (F/Xpk);(b) a transaldolase (Tal);(c) a transketolase (Tkt);(d) a ribose-5-phosphate isomerase (Rpi);(e) a ribulose-5-phosphate epimerase (Rpe);(f) a methanol dehydrogenase (Mdh);(g) a hexulose-6-phosphate synthase (Hps);(h) a hexulose-6-phosphate isomerase (Phi);(i) a dihydroxyacetone synthase (Das); and(j) a fructose-6-phosphate aldolase (Fsa).8. The recombinant microorganism of claim 7 , wherein the microorganism is further engineered to express a phosphotransacetylase.9. (canceled)10. The recombinant microorganism of claim 1 , wherein the phosphoketolase comprises a sequence that is at least 49% identical to SEQ ID NO:2 and has phosphoketolase activity.1114-. (canceled)15. A recombinant prokaryote or yeast microorganism comprising a pathway that produces acetyl-phosphate through carbon rearrangement of E4P and/or G3P and metabolism of a carbon source selected from methane claim 1 , methanol claim 1 , or formaldehyde.1617-. (canceled)18E. coli. The recombinant microorganism of claim 15 , wherein the ...

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

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

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

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

NOVEL AMINO ACIDS BEARING A NORBORNENE MOIETY

The invention relates to a novel amino acid having a norbornene group and polypeptide comprising the novel amino acid compounds. The invention also relates to a method of producing polypeptides comprising a norbornene group and to the use of said polypeptides. 2. The compound of claim 1 , wherein the compound is selected from the group consisting of D- or L-3-norbornene serine claim 1 , D- or L-2-methyl alcohol-3-norbornene serine claim 1 , D- or L-2-methyl-3-norbornene serine claim 1 , D- or L-2-isopropyl alcohol-3-norbornene serine claim 1 , D- or L-2-methyl thiol-3-norbornene serine claim 1 , D- or L-2-butane amine-3-norbornene serine claim 1 , D- or L-2-isopentane-3-norbornene serine claim 1 , D- or L-2-butyric acid-3-norbornene serine claim 1 , D- or L-2-methylpyrrolidine-3-norbornene serine claim 1 , D- or L-2-methyl(propyl)sulfane-3-norbornene serine claim 1 , D- or L-2-1-butylguanidine-3-norbornene serine claim 1 , L-2-propionamide-3-norbornene serine claim 1 , and L-2-butyramide-3-norbornene serine.4. A method for producing compounds of formula I claim 1 , comprising the step of reacting norbornene-2-carboxaldehyde with an amino acid in the presence of threonine aldolase.5. The method according to claim 4 , wherein the amino acid is selected from the group consisting of glycine claim 4 , alanine claim 4 , serine claim 4 , isoleucine claim 4 , leucine claim 4 , threonine claim 4 , glutamic acid claim 4 , proline claim 4 , methionine claim 4 , arginine claim 4 , asparagine claim 4 , glutamine claim 4 , lysine and cysteine.6. The method according to claim 4 , wherein the threonine aldolase is of eukaryotic or prokaryotic origin.7Pseudomonas, Sphingomonas, Azorhizobium, Methylobacterium, Escherichia, Thermotoga, Silicibacter, Paracoccus, Bordetella, ColwelliaSaccharomyces.. The method according to claim 6 , wherein the threonine aldolase is from an organism in a genus selected from the group consisting of claim 6 , and8Pseudomonas putida.. The method according ...

Nanoparticle-attached enzyme cascades for accelerated multistep biocatalysis

A nanoparticle (for example, quantum dot) serves as a substrate for immobilizing enzymes involved in consecutive reactions as a cascade. This results in a significant increase in the rate of catalysis as well as final product yield compared to non-immobilized enzymes. 1. An enzymatic cascade cluster comprising:a plurality of nanoparticles associated together as a cluster, wherein each nanoparticle is bound to a plurality of enzymes configured as an enzymatic cascade wherein the product of a first enzyme is the substrate of a second enzyme and so forth;wherein the enzymatic cascade comprises at least two different enzymes; andwherein the nanoparticles in the cluster are closely associated with one another such that, on average, each nanoparticle is separated from the nearest neighboring nanoparticle by a distance of no more than about one nanoparticle diameter.2. The cluster of claim 1 , wherein the nanoparticle is a quantum dot and the enzymes are bound to the quantum dot via polyhistidine sequences in the enzymes.3. The cluster of claim 1 , wherein said plurality of enzymes comprises pyruvate kinase (PykA) and lactate dehydrogenase (LDH).4. The cluster of claim 1 , wherein said plurality of enzymes comprises glucokinase claim 1 , phosphoglucose isomerase claim 1 , phosphofructokinase claim 1 , fructose-bisphosphate aldolase claim 1 , triose phosphate isomerase claim 1 , glyceraldehyde-3-phosphate dehydrogenase claim 1 , and phosphoglycerate mutase.5. An enzymatic cascade comprising:a nanoparticle bound to a plurality of enzymes configured as an enzymatic cascade wherein the product of a first enzyme is the substrate of a second enzyme and so forth;wherein the enzymatic cascade comprises at least two different enzymes.6. The cascade of claim 5 , wherein the nanoparticle is a quantum dot and the enzymes are bound to the quantum dot via polyhistidine sequences in the enzymes.7. The cascade of claim 5 , wherein said plurality of enzymes comprises pyruvate kinase (PykA) ...

RECOMBINANT MICROORGANISM FOR THE PRODUCTION OF USEFUL METABOLITES

Described are recombinant microorganisms characterized by having phosphoketolase activity, having a diminished or inactivated Embden-Meyerhof-Parnas pathway (EMPP) by inactivation of the gene(s) encoding phosphofructokinase or by reducing phosphofructokinase activity as compared to a non-modified microorganism and having a diminished or inactivated oxidative branch of the pentose phosphate pathway (PPP) by inactivation of the gene(s) encoding glucose-6-phosphate dehydrogenase or by reducing glucose-6-phosphate dehydrogenase activity as compared to a non-modified microorganism. These microorganisms can be used for the production of useful metabolites such as acetone, isobutene or propene. 1. A recombinant microorganism characterized by:a) having phosphoketolase activity;b) (i) having a diminished or inactivated Embden-Meyerhof-Parnas pathway (EMPP) by inactivation of the gene(s) encoding phosphofructokinase or by being genetically modified so as to reduce phosphofructokinase activity as compared to a non-genetically modified microorganism; or(ii) not possessing phosphofructokinase activity; andc) (i) having a diminished or inactivated oxidative branch of the pentose phosphate pathway (PPP) by inactivation of the gene(s) encoding glucose-6-phosphate dehydrogenase or by being genetically modified so as to reduce glucose-6-phosphate dehydrogenase activity as compared to a non-genetically modified microorganism; or(ii) not possessing glucose-6-phosphate dehydrogenase activity, and formate C-acetyltransferase, classified as EC 2.3.1.54;', 'acetate kinase, classified as EC 2.7.2.1', 'L-lactate dehydrogenase, classified as EC 1.1.1.28', 'phosphoenolpyruvate carboxylase, classified as EC 4;1.1.31', 'phosphoenolpyruvate carboxykinase, classified as (ATP) EC 4.1,1.49', 'succinate dehydrogenase, classified as EC 1.3.99.1', 'pyruvate decarboxylase, classified as EC 4.1.1.1', 'acetaldehyde dehydrogenase (acetylating), classified as EC 1.2.1.10', 'glycerol-3-phosphate ...

FRUCTOSE-6-PHOSPHATE ALDOLASE VARIANTS FOR ALDOL CARBOLIGATIONS

The invention provides new and alternative fructose-6-phosphate aldolase (FSA) variants which enable the production of optically active building blocks with high chemoselectivity and stereoselectivity using aldehydes as starting material in aldol carboligation reactions, while avoiding the by-product formation and subsequent reactions. 1. A fructose-6-phosphate aldolase variant comprising:a. an amino acid sequence having at least 80%, preferably at least 90%, more preferably at least 99%, sequence identity with SEQ ID NO: 1, andb. an amino acid substitution at position 6, corresponding to positions 1 to 220 of SEQ ID NO: 1.2. The variant according to claim 1 , wherein the amino acid substitution is by an amino acid selected from the list consisting of: A claim 1 , L claim 1 , N claim 1 , Q claim 1 , S claim 1 , T claim 1 , E claim 1 , H claim 1 , V claim 1 , G claim 1 , I or P claim 1 , preferably by H claim 1 , N claim 1 , Q claim 1 , L claim 1 , T claim 1 , E or A.3. The variant according to any of or claim 1 , wherein said variant further comprises an amino acid substitution at position 26 claim 1 , corresponding to positions 1 to 220 of SEQ ID NO: 1.4. The variant according to claim 3 , wherein the amino acid substitution at position 26 is by an amino acid selected from the list consisting of: V claim 3 , A claim 3 , L claim 3 , I or P claim 3 , preferably by L or I.5. The variant according to any of or claim 3 , wherein said variant further comprises an amino acid substitution at position 165 claim 3 , corresponding to positions 1 to 220 of SEQ ID NO: 1 claim 3 , by the amino acid G.6. The variant according to any one of to claim 3 , wherein said variant comprises the amino acid substitution D6N or D6H.7. A nucleic acid sequence encoding the variant according to any one of to .8. A gene construct comprising the nucleic acid sequence according to .9. A host cell comprising the nucleic acid sequence according to or the gene construct according to .10. Use of the ...

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

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

ALDOLASE, ALDOLASE MUTANT, AND METHOD AND COMPOSITION FOR PRODUCING TAGATOSE BY USING SAME

This disclosure relates to aldolase, an aldolase mutant, and a method and a composition for producing tagatose by using the same. The feature of the disclosure is environment-friendly due to the use of an enzyme acquired from microorganisms, requires only a simple process of enzyme immobilization, uses a low-cost substrate in a substrate compared with a conventional method for producing tagatose and has a remarkably high yield, thereby greatly reducing production costs and maximizing production effects. 12.-. (canceled)3. A method of producing tagatose from fructose , the method comprising:reacting fructose 6-phosphate with fructose 1,6-diphosphate aldolase or a mutant of fructose 1,6-bisphosphate aldolase.4. The method of claim 3 , wherein the fructose 1 claim 3 ,6-diphosphate aldolase is selected from the group consisting of SEQ ID NOS: 1 to 4.511.-. (canceled)12. The method of claim 3 , wherein the mutant includes a substitution at one or more residues at positions of 332 claim 3 , 314 claim 3 , 227 claim 3 , and 62 of fructose aldolased enzyme of SEQ ID NO: 1 claim 3 , wherein the residue at position 332 is substituted from arginine to glutamine claim 3 , the residue at position 314 is substituted from glutamine to alanine claim 3 , the residue at position 227 is substituted from histidine to alanine claim 3 , and the residue at position 62 is substituted from serine to alanine.1314.-. (canceled)15. A mutant enzyme of fructose 1 claim 3 ,6-bisphosphate aldolase claim 3 , which is an enzyme selected from the group consisting of SEQ ID NOS: 1 to 4.16. The mutant enzyme of claim 15 , wherein the mutant enzyme includes a substitution at one or more residues at positions of 332 claim 15 , 314 claim 15 , 227 claim 15 , and 62 of fructose aldolased enzyme of SEQ ID NO: 1 claim 15 , wherein the residue at position 332 is substituted from arginine to glutamine claim 15 , the residue at position 314 is substituted from glutamine to alanine claim 15 , the residue at ...

MICROORGANISMS AND METHODS FOR THE CO-PRODUCTION OF ETHYLENE GLYCOL AND THREE CARBON COMPOUNDS

The present application relates to recombinant microorganisms useful in the biosynthesis of monoethylene glycol (MEG) and one or more three-carbon compounds such as acetone, isopropanol or propene. The MEG and one or more three-carbon compounds described herein are useful as starting material for production of other compounds or as end products for industrial and household use. The application further relates to recombinant microorganisms co-expressing a C2 branch pathway and a C3 branch pathway for the production of MEG and one or more three-carbon compounds. Also provided are methods of producing MEG and one or more three-carbon compounds using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or optionally the products MEG and one or more three-carbon compounds. 2. The recombinant microorganism of claim 1 , wherein the feedstock comprises exogenous glucose.3. The recombinant microorganism of claim 1 , wherein an endogenous or exogenous D-xylose isomerase catalyzes the conversion of D-xylose to D-xylulose.4. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further expresses at least one exogenous nucleic acid molecule encoding a xylose reductase or aldose reductase that catalyzes the conversion of D-xylose to xylitol and at least one exogenous nucleic acid molecule encoding a xylitol dehydrogenase that catalyzes the conversion of xylitol to D-xylulose.5. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises at least one endogenous or exogenous nucleic acid molecule encoding a secondary alcohol dehydrogenase that catalyzes the conversion of acetone to isopropanol.6. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises: at least one endogenous or exogenous nucleic acid molecule encoding a secondary alcohol dehydrogenase that catalyzes the conversion of acetone to isopropanol; and at least one ...

METHODS AND MICROORGANISMS FOR PRODUCING FLAVORS AND FRAGRANCE CHEMICALS

The present disclosure relates to biosynthetic pathways for producing flavor and fragrance chemicals, such as green notes including trans-2-unsaturated aldehydes and lactones. The present disclosure provides methods for producing trans-2-unsaturated aldehydes, delta-lactones, and gamma-lactones. The present disclosure provides pathways for the preparation of trans-2-unsaturated aldehydes, delta-lactones, and gamma-lactones by reacting aldehydes in the presence of aldolases. 167.-. (canceled)69. The non-naturally occurring microorganism of claim 68 , wherein the non-naturally occurring microorganism comprises an increased enzymatic activity of at least one enzyme in the trans-2-unsaturated aldehyde pathway in comparison with the enzymatic activity of the same enzyme in a corresponding unmodified or wild-type microorganism.70. The non-naturally occurring microorganism of claim 68 , wherein the non-naturally occurring microorganism comprises an exogenous nucleic acid that encodes the aldolase.71. The non-naturally occurring microorganism of claim 70 , wherein the aldolase is a pyruvate-dependent aldolase.72. The non-naturally occurring microorganism of claim 68 , wherein the aldolase comprises an amino acid sequence of SEQ ID NO: 11 claim 68 , 12 claim 68 , 13 claim 68 , 14 claim 68 , 15 claim 68 , 16 claim 68 , 17 claim 68 , 18 claim 68 , 19 claim 68 , or 20 claim 68 , an active fragment thereof claim 68 , or a homologue thereof.73. The non-naturally occurring microorganism of claim 72 , wherein the aldolase comprises at least about 50% identity to any one of SEQ ID NOs: 11 claim 72 , 12 claim 72 , 13 claim 72 , 14 claim 72 , 15 claim 72 , 16 claim 72 , 17 claim 72 , 18 claim 72 , 19 claim 72 , or 20.74. The non-naturally occurring microorganism of claim 72 , wherein the aldolase comprises at least about 60% identity to any one of SEQ ID NOs: 11 claim 72 , 12 claim 72 , 13 claim 72 , 14 claim 72 , 15 claim 72 , 16 claim 72 , 17 claim 72 , 18 claim 72 , 19 claim 72 , ...

Production of 2-keto-3-deoxy-d-gluconic acid in filamentous fungi

A recombinant filamentous fungi that includes reduced 2-Keto-3-Deoxy-Gluconate (KDG) aldolase enzyme activity as compared to the filamentous fungi not transformed to have reduced KDG aldolase enzyme activity is provided. Also provided is a method of producing KDG

METHOD FOR THE ENZYMATIC PRODUCTION OF D-ERYTHROSE AND ACETYL PHOSPHATE

Described is a method for the production of D-erythrose and acetyl phosphate comprising the enzymatic conversion of D-fructose into D-erythrose and acetyl phosphate by making use of a phosphoketolase. The produced D-erythrose can further be converted into glycolaldehyde by a method for the production of glycolaldehyde comprising the enzymatic conversion of D-erythrose into glycolaldehyde by making use of an aldolase, wherein said aldolase is a 2-deoxyribose-5-phosphate aldolase (EC 4.1.2.4) or a fructose-bisphosphate aldolase (EC 4.1.2.13). The produced glycolaldehyde can finally be converted into acetyl phosphate by the enzymatic conversion of the thus produced glycolaldehyde into acetyl phosphate by making use of a phosphoketolase or a sulfoacetaldehyde acetyltransferase. 1. A method of producing D-erythrose and acetyl phosphate comprising enzymatically converting D-fructose into D-erythrose and acetyl phosphate by a phosphoketolase.2. The method of claim 1 , wherein the phosphoketolase is(a) a phosphoketolase (EC 4.1.2.9), or(b) a fructose-6-phosphate phosphoketolase (EC 4.1.2.22).3. A method of producing glycolaldehyde comprising enzymatically converting D-erythrose into glycolaldehyde by an aldolase claim 1 , wherein said aldolase is a 2-deoxyribose-5-phosphate aldolase (EC 4.1.2.4) or a fructose-bisphosphate aldolase (EC 4.1.2.13).4. The method of claim 3 , wherein the method further comprises:(a) enzymatically converting D-fructose into D-erythrose and acetyl phosphate by a phosphoketolase; and{'claim-ref': {'@idref': 'CLM-00003', 'claim 3'}, '(b) enzymatically converting D-erythrose into glycolaldehyde according to a method of .'}5. The method of claim 3 , wherein the method further comprises:enzymatically converting glycolaldehyde into acetyl phosphate by a phosphoketolase or a sulfoacetaldehyde acetyltransferase (EC 2.3.3.15).6. The method of claim 5 , wherein the phosphoketolase is(i) a phosphoketolase (EC 4.1.2.9), or(ii) a fructose-6-phosphate ...

Utilization of phosphoketolase in the production of mevalonate, isoprenoid precursors, and isoprene

The invention provides for methods for the production of mevalonate, isoprene, isoprenoid precursor molecules, and/or isoprenoids in cells via the heterologous expression of phosphoketolase enzymes.

GLUCOSE METABOLISM WITH MOLECULAR PURGE VALVE

Provided is an engineered pathway that can function in a cell-free system, cellular system or a combination thereof to convert a sugar to a chemical or biofuel. 1. A recombinant , artificial or engineered metabolic pathway comprising a plurality of enzymatic steps that converts a substrate to acetyl-phosphate , pyruvate , glyceraldehyde-3-phosphate , or acetyl-CoA , wherein the pathway includes an unbalanced production and utilization of a co-factor , the pathway comprising a non-naturally occurring purge valve pathway that recycles the co-factor , wherein the purge valve pathway comprises an enzyme that uses the co-factor to convert a metabolite to an intermediate or product in one or more of the plurality of enzymatic steps.2. The recombinant claim 1 , artificial or engineered pathway of claim 1 , wherein the co-factors are oxidizing/reducing co-factors.3. The recombinant claim 2 , artificial or engineered pathway of claim 2 , wherein the oxidizing/reducing co-factors are NAD/NADH claim 2 , NADP/NADPH or FAD/FADH.4. The recombinant claim 3 , artificial or engineered pathway of claim 3 , wherein a first cofactor comprises NAD/NADH and a second cofactor comprises NADP/NADPH.5. The recombinant claim 1 , artificial or engineered pathway of claim 1 , wherein the purge valve pathway comprises an NADH oxidase.6. The recombinant claim 5 , artificial or engineered pathway of claim 5 , wherein the NADH oxidase is a NoxE or homolog thereof.7. The recombinant claim 6 , artificial or engineered pathway of claim 6 , wherein the NADH oxidase comprises a sequence that is at least 50% identical to SEQ ID NO:18.8. The recombinant claim 1 , artificial or engineered pathway of claim 1 , wherein the pathway carries out the following:(i) converts glucose to glucose-6-phosphate;(ii) converts glucose-6-phosphate to 6-phospho-D-glucono-1,5-lactone;(iii) converts 6-phospho-D-glucono-1,5-lactone to 6-phospho-D-gluconate;(iv) converts 6-phospho-D-gluconate to ribulose-5-phosphate;(v) ...

CELL-FREE AND MINIMIZED METABOLIC REACTION CASCADES FOR THE PRODUCTION OF CHEMICALS

Provided are enzymatic processes for the production of chemicals like ethanol from carbon sources like glucose, in particular, a process for the production of a target chemical is disclosed using a cell-free enzyme system that converts carbohydrate sources to the intermediate pyruvate and subsequently the intermediate pyruvate to the target chemical wherein a minimized number of enzymes and only one cofactor is employed. 2. The process of claim 1 , wherein the temperature of the process is maintained in a range from 40-80° C.3. The process of claim 1 , wherein the process is maintained at the given temperature for at least 1 hour.4. The process of claim 1 , step (1) further comprising the use of a single dehydrogenase for the oxidation of glucose and/or galactose to gluconate and/or galactonate.5. The process of claim 1 , step (2) further comprising the conversion of gluconate to 2-keto-3-deoxygluconate and of 2-keto-3-deoxygluconate to glyceraldehyde and pyruvate and/or the conversion of galaconate to 2-keto-3-deoxygalactonate and of 2-keto-3-deoxygalactonate to glyceraldehyde and pyruvate.6. The process of claim 1 , step (2) further comprising the use of dehydroxy acid dehydratase and keto-3-deoxygluconate aldolase.7. The process of claim 1 , step (3) further comprising the use of a dehydrogenase for the oxidation of glyceraldehyde to glycerate.8. The process of claim 1 , wherein steps (1) and (3) are carried out with the use of a single dehydrogenase.9. The process of claim 1 , wherein no net production of ATP occurs.10. The process of claim 1 , wherein no ATPase or arsenate is added.11. The process of claim 1 , wherein said process occurs without ATP and/or ADP as cofactors.12. The process of claim 1 , wherein the single cofactor is selected from the group consisting of NAD/NADH claim 1 , NADP/NADPH claim 1 , and FAD/FADH2.13. The process of claim 11 , wherein the single cofactor is NAD/NADH.14. The process of claim 1 , wherein the enzyme activity of each ...

Utilization of phosphoketolase in the production of mevalonate, isoprenoid precursors, and isoprene

The invention provides for methods for the production of mevalonate, isoprene, isoprenoid precursor molecules, and/or isoprenoids in cells via the heterologous expression of phosphoketolase enzymes.

CARBON DIOXIDE FIXATION VIA BYPASSING FEEDBACK REGULATION

Genetically engineered cells and methods are presented that allow for the production of various value products from CO. Contemplated cells have a CBB cycle that is genetically modified such that two molecules of COfixed in the CBB cycle can be withdrawn from the modified CBB cycle as a single C2 compound. In contemplated aspects a CBB cycle includes an enzymatic activity that generates the single C2 compound from a compound of the CBB cycle, while further modifications to the CBB cycle will not introduce additional recombinant enzymatic activity/activities outside the already existing catalytic activities in the CBB cycle. 1. A metabolically engineered cell having a native CBB cycle , comprising: 'wherein the first enzyme is a phosphoketolase enzyme (EC 4.1.2.9), that utilizes an intermediate of the CBB pathway as a substrate, and generates a first acetyl phosphate product; and the second enzyme is a phosphoribulokinase enzyme (EC 2.7.1.1) that is overexpressed in the genetically modified organism in an amount to achieve a phosphoribulokinase activity level that is higher than the native phosphoribulokinase activity level of the organism, and the second enzyme utilizes ribulose-5-phosphate to produce ribulose-1,5-bisphosphate.', 'a recombinant nucleic acid comprising a nucleic acid sequence encoding a first enzyme and a second enzyme;'}2. The metabolically engineered cell of claim 1 , wherein the cell is a bacterial cell.3Ralstonia.. The metabolically engineered cell of claim 2 , wherein the bacterial cell belongs to the genus4. The metabolically engineered cell of claim 1 , wherein production of phosphoenolpyruvate in the genetically modified organism claim 1 , when grown under nitrogen depletion claim 1 , is below a feedback inhibitory concentration for the CBB cycle.5. The metabolically engineered cell of claim 3 , that comprises a plasmid comprising an alsS-ilvC-ilvD operon.6. The metabolically engineered cell of claim 3 , that comprises a plasmid comprising a ...

Method of Producing Lipid

A method of improving photosynthetic ability of an alga, containing enhancing expression of a transketolase and a fructose-1,6-bisphosphate aldolase. 1. A method of improving photosynthetic ability of an alga , comprising enhancing expression of a transketolase and a fructose-1 ,6-bisphosphate aldolase.2. A method of producing lipids , comprising the steps of:culturing an alga in which expression of a transketolase and expression of a fructose-1,6-bisphosphate aldolase are enhanced, andproducing fatty acids or lipids containing the same as components.3. A method of improving lipid productivity , comprisingenhancing expression of a transketolase and a fructose-1,6-bisphosphate aldolase to improve productivity of fatty acids or lipids containing the same as components produced in an algal cell.4. (canceled)5. The method according to claim 2 , wherein the fatty acids or lipids containing the same as components are palmitic acids or lipids containing the same as components.6. The method according to claim 1 , wherein the transketolase is the following protein (A) or (B) claim 1 , and the fructose-1 claim 1 ,6-bisphosphate aldolase is the following protein (C) or (D):(A) a protein consisting of the amino acid sequence set forth in SEQ ID NO: 1;(B) a protein consisting of an amino acid sequence having 60% or more identity with the amino acid sequence of the protein (A), and having transketolase activity;(C) a protein consisting of the amino acid sequence set forth in SEQ ID NO: 3; and(D) a protein consisting of an amino acid sequence having 60% or more identity with the amino acid sequence of the protein (C), and having fructose-1,6-bisphosphate aldolase activity.7. The method according to claim 1 ,wherein expression of the following protein (E) or (F) is enhanced in the alga:(E) a protein consisting of the amino acid sequence set forth in SEQ ID NO: 7; or(F) a protein consisting of an amino acid sequence having 60% or more identity with the amino acid sequence of the ...

Polypeptide Assemblies and Methods for the Production Thereof

The application discloses multimeric assemblies including multiple oligomeric substructures, where each oligomeric substructure includes multiple proteins that self-interact around at least one axis of rotational symmetry, where each protein includes one or more polypeptide-polypeptide interface (“O interface”); and one or more polypeptide domain that is capable of effecting membrane scission and release of an enveloped multimeric assembly from a cell by recruiting the ESCRT machinery to the site of budding by binding to one or more proteins in the eukaryotic ESCRT complex (“L domain”); and where the multimeric assembly includes one or more subunits comprising one or more polypeptide domain that is capable of interacting with a lipid bilayer (“M domain”), as well as membrane-enveloped versions of the multimeric assemblies. 1. A multimeric assembly , comprising a plurality of oligomeric substructures , wherein each oligomeric substructure comprises a plurality of proteins that self-interact around at least one axis of rotational symmetry , wherein each protein comprises:(a) one or more polypeptide-polypeptide interface (“O interface”); and(b) one or more polypeptide domain that is capable of effecting membrane scission and release of an enveloped multimeric assembly from a cell by recruiting the ESCRT machinery to the site of budding by binding directly or indirectly to one or more ESCRT or ESCRT-associated proteins (“L domain”);wherein one or more protein in the multimeric assembly comprises one or more polypeptide domain that is capable of interacting with a lipid bilayer (“M domain”);wherein the M domain, L domain, and O interface are not each present in a single naturally occurring protein, wherein the plurality of oligomeric substructures interact with each other at the one or more O interfaces.2. The multimeric assembly of claim 1 , wherein at least one protein in each oligomeric structure comprises one or more M domain claim 1 , or wherein each protein ...

Recombinant yeast and substance production method using the same

Substance productivity is improved by introducing a metabolic pathway for synthesis of acetyl-CoA or acetic acid from glucose-6-phosphate into yeast. Acetic acid productivity, acetyl-CoA productivity, and productivity of a substance made from acetyl-CoA-derived are improved by attenuating genes involved in the glycolytic system of yeast and introducing a phosphoketolase gene into the yeast.

INCREASED ALCOHOL PRODUCTION FROM YEAST PRODUCING AN INCREASED AMOUNT OF ACTIVE CRZ1 PROTEIN

Described are compositions and methods relate to modified yeast that, in addition to native endogenous CRZ1, produces a modified CRZ1 transcriptional activator involved in the calcineurin stress response pathway. Such yeast is well suited for use in fuel alcohol production to increase yield. 1. Modified yeast cells derived from parental yeast cells , the modified cells comprising a genetic alteration that causes the cells to produce an increased amount of active CRZ1 polypeptides compared to the parental cells , wherein the modified cells produce during fermentation an increased amount of alcohol compared to the amount of alcohol produced by the parental cells under identical fermentation conditions.2. The modified cells of claim 1 , wherein the active CRZ1 polypeptides exhibit reduced phosphorylation compared to native CRZ1 polypeptides under identical fermentation conditions.3. The modified cells of or claim 1 , wherein the active CRZ1 polypeptides include a reduced number of amino acid residues capable of phosphorylation compared to the amino acid residues in native CRZ1 polypeptides.4. The modified cells of claim 3 , wherein the active CRZ1 polypeptides include a reduced number of serine residues capable of phosphorylation compared to the amino acid residues in native CRZ1 polypeptides.5. The modified cells of any of - claim 3 , wherein the amount of increase in the expression of the modified CRZ1 mutant polypeptides is at least about 500% claim 3 , at least 1 claim 3 ,000% claim 3 , at least 1 claim 3 ,500% claim 3 , or even at least 2 claim 3 ,000% claim 3 , compared to the level expression of native CRZ1 polypeptides in the parental cells grown under equivalent conditions.6. The modified cells of any of - claim 3 , wherein the cells further comprising one or more genes of the phosphoketolase pathway.7. The modified cells of claim 6 , wherein the genes of the phosphoketolase pathway are selected from the group consisting of phosphoketolase claim 6 , ...

Vanillin Synthase

The invention relates to methods for producing vanillin and related compounds. The methods involve use of a vanillin synthase capable of catalyzing side chain cleavage of ferulic acid to form vanillin. The invention also relates to host organisms expressing such vanillin synthases useful in the methods.

MICROORGANISMS AND METHODS FOR THE CO-PRODUCTION OF ETHYLENE GLYCOL AND THREE CARBON COMPOUNDS

The present application relates to recombinant microorganisms useful in the biosynthesis of monoethylene glycol (MEG) and one or more three-carbon compounds such as acetone, isopropanol or propene. The MEG and one or more three-carbon compounds described herein are useful as starting material for production of other compounds or as end products for industrial and household use. The application further relates to recombinant microorganisms co-expressing a C2 branch pathway and a C3 branch pathway for the production of MEG and one or more three-carbon compounds. Also provided are methods of producing MEG and one or more three-carbon compounds using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or optionally the products MEG and one or more three-carbon compounds. 14.-. (canceled)5. A recombinant microorganism capable of co-producing monoethylene glycol (MEG) and acetone from a feedstock comprising exogenous D-xylose , wherein the recombinant microorganism expresses one or more of the following:(a) at least one endogenous or exogenous nucleic acid molecule encoding a D-tagatose 3-epimerase that catalyzes the conversion of D-xylulose to D-ribulose;(b) at least one endogenous or exogenous nucleic acid molecule encoding a D-ribulokinase that catalyzes the conversion of D-ribulose from (a) to D-ribulose-1-phosphate;(c) at least one endogenous or exogenous nucleic acid molecule encoding a D-ribulose-1-phosphate aldolase that catalyzes the conversion of D-ribulose-1-phosphate from (b) to glycolaldehyde and dihydroxyacetonephosphate (DHAP);(d) at least one endogenous or exogenous nucleic acid molecule encoding a glycolaldehyde reductase that catalyzes the conversion of glycolaldehyde from (c) to MEG;(e) at least one exogenous nucleic acid molecule encoding a thiolase that catalyzes the conversion of acetyl-CoA to acetoacetyl-CoA;(f) at least one endogenous or exogenous nucleic acid molecule encoding an acetate: ...

Enzymatic production of an acyl phosphate from a 2-hydroxyaldehyde

Described is a method for the enzymatic production of an acyl phosphate from a 2-hydroxyaldehyde using a phosphoketolase or a sulfoacetaldehyde acetyltransferase.

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

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

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

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

GENOMIC ENGINEERING OF BIOSYNTHETIC PATHWAYS LEADING TO INCREASED NADPH

The disclosure relates to host cells having altered NADPH availability, allowing for increased production of compounds produced using NADPH, and methods of use thereof. NADPH availability is altered by one or more of: expressing an altered GAPDH, expressing a variant glutamate dehydrogenase (gdh), aspartate semialdehyde dehydrogenase (asd), dihydropicolinate reductase (dapB), and meso-diaminopimelate dehydrogenase (ddh), expressing a novel nicotinamide nucleotide transhydrogenase, expressing a novel threonine aldolase, and expressing or modulating the expression of a pyruvate carboxylase in the host cells. 1. A method of improving a microbial cell's ability to produce a compound produced using NADPH , the method comprising altering the cell's available NADPH.2. The method of claim 1 , wherein the available NADPH is altered by expressing a modified Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in the cell claim 1 , wherein the modified GAPDH is modified such that its coenzyme specificity is broadened.3. The method of claim 1 , wherein the cell's available NADPH is altered by expressing claim 1 , in the microbial cell claim 1 , a variant enzyme of one or more of the enzymes glutamate dehydrogenase (gdh) claim 1 , aspartate semialdehyde dehydrogenase (asd) claim 1 , dihydropicolinate reductase (dapB) claim 1 , and meso-diaminopimelate dehydrogenase (ddh) claim 1 , wherein the variant enzyme exhibits dual specificity for coenzymes NADH and NADPH.4. (canceled)5. The method of claim 1 , wherein the microbial cell is a bacterial cell.6CorynebacteriumEscherichiaBacillusGeobacillus. The method of claim 5 , wherein the bacterial cell is from a bacteria selected from the group consisting of sp. claim 5 , sp. claim 5 , sp. or sp.7Corynebacterium glutamicumEscherichia coli.. The method of claim 5 , wherein the bacteria is or823-. (canceled)24. The method of claim 1 , wherein the compound is selected from Table 2.25. The method of claim 24 , wherein the compound is L-lysine or ...

A FRUCTOSE-C4-EPIMERASE AND PREPARATION METHOD FOR PRODUCING TAGATOSE USING THE SAME

The present disclosure relates to a composition for producing tagatose comprising a protein having fructose-4-epimerase activity and a method for producing tagatose using the same. 1Caldilinea. A composition for producing tagatose , comprising at least one of tagatose-bisphosphate aldolase derived from sp.; a microorganism expressing the tagatose-bisphosphate aldolase; and a culture of the microorganism.2. The composition of claim 1 , wherein the composition further comprises fructose.3. The composition of claim 2 , wherein the fructose is D-fructose.4. The composition of claim 1 , wherein the tagatose-bisphosphate aldolase has an amino acid sequence of SEQ ID NO: 1 or an amino acid sequence having an identity of 85% or more thereto.5. The composition of claim 1 , wherein the composition further comprises a metal ion or a metal salt.6. The composition of claim 5 , wherein the metal is at least one selected from the group consisting of manganese and nickel.7Caldilinea. A method for producing tagatose claim 5 , comprising converting fructose to tagatose by contacting fructose with at least one of tagatose-bisphosphate aldolase derived from sp.; a microorganism expressing the tagatose-bisphosphate aldolase; and a culture of the microorganism.8. The method of claim 7 , wherein the contact is performed at a pH of 7.0 to 9.0 and at a temperature of 45° C. to 80° C. for 0.5 hours to 24 hours.9. The method of claim 7 , wherein the conversion of fructose is performed in the presence of a metal ion or a metal salt.10. The method of claim 8 , wherein the metal is at least one selected from the group consisting of manganese and nickel.11. The method of claim 7 , wherein the tagatose-bisphosphate aldolase has an amino acid sequence of SEQ ID NO: 1 or an amino acid sequence having an identity of 85% or more thereto. The present disclosure relates to a composition for producing tagatose including a protein having fructose-4-epimerase activity and a method for producing tagatose ...

NOVEL FRUCTOSE-4-EPIMERASE AND TAGATOSE PRODUCTION METHOD USING SAME

Provided are a tagatose-bisphosphate aldolase variant having tagatose conversion activity, and a method of preparing tagatose using the same. 1. A fructose-4-epimerase , wherein one or more amino acid residues are substituted in fructose-4-epimerase including an amino acid sequence of SEQ ID NO:1.2. The fructose-4-epimerase variant of claim 1 , wherein the fructose-4-epimerase variant has substitution of another amino acid for any one or more selected from the group consisting of amino acids at positions of 8 claim 1 , 20 claim 1 , 23 claim 1 , 25 claim 1 , 26 claim 1 , 29 claim 1 , 45 claim 1 , 51 claim 1 , 53 claim 1 , 63 claim 1 , 86 claim 1 , 91 claim 1 , 97 claim 1 , 110 claim 1 , 133 claim 1 , 144 claim 1 , 146 claim 1 , 151 claim 1 , 155 claim 1 , 167 claim 1 , 172 claim 1 , 173 claim 1 , 174 claim 1 , 181 claim 1 , 191 claim 1 , 239 claim 1 , 263 claim 1 , 266 claim 1 , 285 claim 1 , 294 claim 1 , 298 claim 1 , 308 claim 1 , 315 claim 1 , 316 claim 1 , 317 claim 1 , 323 claim 1 , 336 claim 1 , 347 claim 1 , 359 claim 1 , 367 claim 1 , 385 claim 1 , 386 claim 1 , 388 claim 1 , 389 claim 1 , 410 claim 1 , 414 claim 1 , and 417 in the amino acid sequence of SEQ ID NO: 1.3. The fructose-4-epimerase variant of claim 2 , wherein the another amino acid is selected from the group consisting of glycine claim 2 , alanine claim 2 , arginine claim 2 , valine claim 2 , leucine claim 2 , methionine claim 2 , isoleucine claim 2 , threonine claim 2 , asparagine claim 2 , glutamine claim 2 , proline claim 2 , serine claim 2 , tryptophan claim 2 , phenylalanine claim 2 , histidine claim 2 , cysteine claim 2 , tyrosine claim 2 , lysine claim 2 , aspartic acid claim 2 , and glutamic acid.4. A polynucleotide encoding the fructose-4-epimerase variant of .5. A vector comprising the polynucleotide of .6. A microorganism comprising the fructose-4-epimerase variant of ; a polynucleotide encoding the fructose-4-epimerase variant; or a vector including the polynucleotide.7. A ...

ACETIC ACID CONSUMING STRAIN

The invention describes a process for the production of ethanol from a composition comprising glucose and between 50 μM and 100 mM acetic acid, said process comprising fermenting said composition in the presence of a recombinant yeast which is capable to convert acetic acid anaerobically; maintaining the amount of undissociated acetic acid at a value of at least 50 μM; and recovering the ethanol. Said process is useful for both starch and cellulosic based, acetic acid containing hydrolysates and advantageously results in a greater consumption of acetic acid and thus higher ethanol yields. 1. Process for production of ethanol from a composition comprising glucose and between 50 μM and 100 mM acetic acid , said process comprising:fermenting said composition in the presence of a recombinant yeast which is capable to convert acetic acid anaerobically;maintaining the amount of undissociated acetic acid at a value of at least 50 μM; andrecovering the ethanol.2. Process according to wherein said maintaining the amount of dissociated acetic acid to a value of at least 50 μM comprises:monitoring the amount of undissociated acetic acid in the composition, and if the amount of undissociated acetic acid drops below 50 μM;adding acid to the composition until the amount of undissociated acetic acid reaches a value of at least 50 μM, optionally by adding an acid.3. Process according to wherein said maintaining the amount of dissociated acetic acid to a value of at least 50 μM comprises:monitoring the amount of undissociated acetic acid in the composition, and if the amount of undissociated acetic acid approaches 50 μM, but before the amount drops below 50 μM:adding acid to the composition until the amount of undissociated acetic acid reaches a value of above 50 μM, optionally by adding an acid.4. Process according to wherein the recombinant yeast comprises:a nucleic acid sequence encoding an enzyme having acetylating acetaldehyde dehydrogenase activity (EC 1.2.1.10 or EC 1.1.1.2); ...

HIGH-LEVEL PRODUCTION OF DIACETYL IN A METABOLICALLY ENGINEERED LACTIC ACID BACTERIUM

The present invention provides a genetically modified lactic acid bacterium capable of producing diacetyl under aerobic conditions. Additionally the invention provides a method for producing diacetyl using the genetically modified lactic acid bacterium under aerobic conditions in the presence of a source of iron-containing porphyrin and a metal ion selected from Fe, Fe and Cu2+. The lactic acid bacterium is genetically modified by deletion of those genes in its genome that encode polypeptides having lactate dehydrogenase (E.C 1.1.1.27/E.C.1.1.1.28); α-acetolactate decarboxylase (E.C 4.1.1.5); water-forming NADH oxidase (E.C. 1.6.3.4); phosphotransacetylase (E.C.2.3.1.8) activity; and optionally devoid of or deleted for genes encoding polypeptides having diacetyl reductase ((R)-acetoin forming; EC: 1.1.1.303); D-acetoin reductase; butanediol dehydrogenase ((R,R)-butane-2,3-diol forming; E.C. 1.1.1.4/1.1.1.-) and alcohol dehydrogenase (E.C. 1.2.1.10) activity. The invention provides for use of the genetically modified lactic acid bacterium for the production of diacetyl and a food product. 1. A genetically modified lactic acid bacterium for production of diacetyl , wherein the genome of said lactic acid bacterium is deleted for genes or lacks genes encoding polypeptides having an enzymatic activity of:a. lactate dehydrogenase (E.C.1.1.1.27 or E.C.1.1.1.28)b. α-acetolactate decarboxylase (E.C. 4.1.1.5)c. phosphotransacetylase (E.C.2.3.1.8) andd. NADH oxidase (E.C. 1.6.3.4); andwherein said microorganism is devoid of transgenes encoding polypeptides having an enzymatic activity of:e. a diacetyl reductase (E.C.1.1.1.304) andf. a L-butanediol dehydrogenase (E.C. 1.1.1.76)2. A genetically modified lactic acid bacterium according to claim 1 , wherein the genome of said lactic acid bacterium is additionally deleted for one or more genes or lacks one or more genes encoding polypeptides having an enzymatic activity selected from the group of:a. alcohol dehydrogenase (E.C. 1.2. ...

PICHIA CIFERRII CELLS AND USES THEREOF

The invention relates to genetically modified cells, to the use thereof and to a method of producing sphingoid bases and sphingolipids. 1Pichia ciferriiPichia ciferrii. A genetically modified cell , wherein said genetically modified cell has , compared to its wild type , a reduced activity of at least one of the enzymes encoded by the intron-free nucleic acid sequences selected from the two groups A) and B) consisting of:A) SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11; andB) a sequence which is at least 80% identical to any of the sequences SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11.2Pichia ciferrii. The genetically modified cell according to , characterized in that wherein said reduction of the enzymic enzymatic activity is achieved by modifying a gene comprising any of the nucleic acid sequences specified in , wherein the modification is selected from the group consisting of:insertion of foreign DNA into the gene,deletion of at least a portion of the gene,point mutations in the gene sequence,exposing the gene to the influence of RNA interference, andreplacement of a portion of the gene with foreign DNA.3Pichia ciferrii. The genetically modified cell according to claim 2 , wherein said foreign DNA is a selection marker gene that can be removed without leaving a trace and which leaves a deletion in the target gene.4Pichia ciferriiPichia ciferrii. The genetically modified cell according to claim 1 , wherein the cell is obtained from strains selected from the group consisting of{'i': 'Pichia ciferrii', 'NRRL Y-1031 F-60-10; and'}{'i': 'Pichia ciferrii', 'CS.PCAPro2.'}5Pichia ciferrii. The genetically modified cell according to claim 1 , characterized in that the cell further has claim 1 , compared to its wild type claim 1 , an increased enzymatic activity of at least one of the enzymes selected from the group consisting of;{'sub': 1', '1, 'enzyme E, wherein enzyme Ecatalyses the reaction of serine and palmitoyl-CoA ...

NON-NATURAL MICROBIAL ORGANISMS WITH IMPROVED ENERGETIC EFFICIENCY

The invention provides non-natural microbial organisms containing enzymatic pathways and/or metabolic modifications for enhancing carbon flux through acetyl-CoA, or oxaloacetate and acetyl-CoA. Embodiments of the invention include microbial organisms having a pathway to acetyl-CoA and oxaloacetate that includes phosphoketolase (a PK pathway). The organisms also have either (i) a genetic modification that enhances the activity of the non-phosphotransferase system (non-PTS) for sugar uptake, and/or (ii) a genetic modification(s) to the organism's electron transport chain (ETC) that enhances efficiency of ATP production, that enhances availability of reducing equivalents or both. The microbial organisms can optionally include (iii) a genetic modification that maintains, attenuates, or eliminates the activity of a phosphotransferase system (PTS) for sugar uptake. The enhanced carbon flux through acetyl-CoA and oxaloacetate can be used for production of a bioderived compound, and the microbial organisms can further include a pathway capable of producing the bioderived compound. 1. A non-natural microbial organism capable of producing acetyl-CoA , or acetyl-CoA and oxaloacetate , the organism comprising:(a) a pathway comprising phosphoketolase for producing acetyl-CoA (PK pathway);(b) a non-phosphotransferase system (non-PTS) for sugar uptake comprising a modification to increase non-PTS activity; and optionally(c) a modification that attenuates or eliminates a PTS activity.2. The non-natural organism of further comprising one or more modification(s) to the organism's electron transport chain to enhance efficiency of ATP production claim 1 , to enhance availability of reducing equivalents claim 1 , or both.3. A non-natural microbial organism capable of producing acetyl-CoA claim 1 , or acetyl-CoA and oxaloacetate claim 1 , the organism comprising:(a) a pathway comprising phosphoketolase for producing acetyl-CoA (PK pathway); and(b) one or more modification(s) to the ...

DEGRADATION PATHWAY FOR PENTOSE AND HEXOSE SUGARS

The present application relates to recombinant microorganisms useful in the biosynthesis of monoethylene glycol (MEG) or glycolic acid (GA), or MEG and one or more co-product, from one or more pentose and/or hexose sugars. Also provided are methods of producing MEG (or GA), or MEG (or GA) and one or more co-product, from one or more pentose and/or hexose sugars using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or the products MEG (or GA), or MEG and one or more co-product. 1. A recombinant microorganism expressing at least one enzyme having pentose-phosphate aldolase activity wherein said microorganism produces one or more products derived from glyceraldehyde-3-phosphate (G3P) and glycolaldehyde from one or more pentose and/or hexose sugars via a pentose-phosphate intermediate; wherein the one or more product is selected from monoethylene glycol (MEG) and glycolic acid (GA) wherein the pentose-phosphate intermediate is D-ribose-5-phosphate , D-ribulose-5-phosphate or D-xylulose-5-phosphate and wherein the enzyme have D-ribose-5-phosphate , D-ribulose-5-phosphate or D-xylulose-5-phosphate aldolase activity.23.-. (canceled)4. The recombinant microorganism of claim 1 , wherein the recombinant microorganism co-produces monoethylene glycol (MEG) or glycolic acid (GA) and one or more co-products claim 1 , wherein the one or more co-products are selected from acetone claim 1 , isopropanol claim 1 , propene claim 1 , L-serine claim 1 , glycine claim 1 , monoethanolamine (MEA) claim 1 , ethylenediamine claim 1 , or a combination thereof.5. (canceled)6E. coli;. The recombinant microorganism of claim 1 , wherein the microorganism comprises expression of at least one enzyme having transketolase activity claim 1 , wherein the at least one enzyme having transketolase activity is encoded by an amino acid sequence having at least 70% sequence identity claim 1 , at least 80% sequence identity claim 1 , or at least 90% ...

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

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

RECOMBINANT CELL AND METHOD FOR PRODUCING ISOPRENE

An object of the present invention is to provide a series of techniques for producing isoprene from methanol or the like. Provided is a recombinant cell prepared by introducing a gene encoding isoprene synthase, into a host cell which is a methylotroph, wherein the gene 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 methane, methanol, methylamine, formic acid, formaldehyde, and formamide. Preferably, it has at least one C1 carbon assimilating pathway selected from the group consisting of a serine pathway, a ribulose monophosphate pathway, and a xylulose monophosphate pathway as a fixing pathway of formaldehyde. Also provided is a method for producing isoprene using the recombinant cell. 139-. (canceled)40. A recombinant cell prepared by introducing a gene encoding isoprene synthase , into a host cell which is methylotroph , whereinthe gene is expressed in the host cell, andthe recombinant cell is capable of producing isoprene from at least one C 1 compound selected from the group consisting of methane, methanol, methylamine, formic acid, formaldehyde, and formamide.41. The recombinant cell according to claim 40 , having at least one C 1 carbon assimilating pathway selected from the group consisting of a serine pathway claim 40 , a ribulose monophosphate pathway claim 40 , and a xylulose monophosphate pathway as a fixing pathway of formaldehyde.42. The recombinant cell according to claim 40 , wherein a gene encoding 3-hexulose-6-phosphate synthase and a gene encoding 6-phospho-3-hexuloisomerase are further introduced claim 40 , and the genes are expressed in the host cell.43. The recombinant cell according to claim 40 ,wherein the host cell has isopentenyl diphosphate synthesis ability by a mevalonate pathway, andwherein a gene encoding at least one enzyme acting in a mevalonate pathway and/or a gene encoding a group of enzymes acting in a non-mevalonate ...

NOVEL BACTERIAL STRAINS FOR THE PRODUCTION OF VANILLIN

The invention relates to an strain derived from the Zyl 926 strain having at least one additional copy of the FCS and ECH genes encoding feruloyl-CoA-synthetase and enoyl-CoA-hydratase/aldolase integrated at the integration site of the φC31 phage. 1AmycolatopsisAmycolatopsis. An strain derived from the Zyl 926 strain filed with the CABI Bioscience under the number IMI 390106 on 2 Mar. 2003 , characterized in that it comprises at least one additional copy of the FCS gene encoding feruloyl-coA-synthetase having the protein sequence SEQ ID NO: 1 or any sequence at least 80% , preferably at least 90% , and more preferably at least 95% identical to SEQ ID NO sequence: 1 , and at least one additional copy of the ECH gene encoding Enoyl-coA Hydratase/Aldolase having the protein sequence SEQ ID NO: 2 or any sequence at least 80% , preferably at least 90% , and more preferably at least 95% identical to SEQ ID NO sequence: 2 and in that said additional copies of the FCS and ECH genes are specifically incorporated at the φC31 phage integration site.2Amycolatopsis. An strain according to claim 1 , characterized in that said additional copies of the FCS and ECH genes are placed under the control of strong ermE* promoter associated with a ribosome combining site.3AmycolatopsisAmycolatopsis. A process for obtaining an strain derived from the Zyl 926 strain filed with the CABI Bioscience under the number IMI 390106 on 2 Mar. 2003 claim 1 , comprising at least one additional copy of the FCS gene encoding feruloyl-coA-synthetase having the sequence SEQ ID NO: 1 or any sequence at least 80% claim 1 , preferably at least 90% claim 1 , and more preferably at least 95% identical to SEQ ID NO sequence: 1 claim 1 , and the ECH gene encoding the Enoyl-coA Hydratase/Aldolase having the sequence SEQ ID NO sequence: 2 or any sequence at least 80% claim 1 , preferably at least 90% claim 1 , and more preferably at least 95% identical to SEQ ID NO sequence: 2 and of which said additional copies ...

Methods and Systems for 1-Butanol Production

A combination of an electrochemical device for delivering reducing equivalents to a cell, and engineered metabolic pathways within the cell capable of utilizing the electrochemically provided reducing equivalents is disclosed. Such a combination allows the production of commodity chemicals by fermentation to proceed with increased carbon efficiency. 1. A system for 1-butanol production , comprising:an electrochemical bioreactor module for providing reducing equivalents;an engineered cell for receiving and using the reducing equivalents to produce 1-butanol, wherein the engineered cell comprises exogenously introduced enzymes selected from pyruvate:formate lyase (Pfl, EC 2.3.1.54), formaldehyde dehydrogenase (Fld, EC 1.2.1.46), hexulose-6-phosphate synthase (HPS, EC 4.1.2.43), and 6-phospho-3-hexuloisomerase (HPI, EC 5.3.1.27), and wherein in the engineered cell the endogenous pyruvate dehydrogenase (Pdh, EC 1.2.4.1) has been disabled, deleted or otherwise rendered non-functional.2. The system of wherein the engineered cell comprises exogenously introduced pyruvate:formate lyase (Pfl claim 1 , EC 2.3.1.54) claim 1 , formaldehyde dehydrogenase (Fld claim 1 , EC 1.2.1.46) claim 1 , hexulose-6-phosphate synthase (HPS claim 1 , EC 4.1.2.43) claim 1 , and 6-phospho-3-hexuloisomerase (HPI claim 1 , EC 5.3.1.27).3. The system of wherein the engineered cell further comprises exogenously introduced acetyl-CoA acetyltransferase (AtoB claim 1 , EC 2.3.1.9) claim 1 , 3-hydroxybutyryl-CoA dehydrogenase (Hbd claim 1 , EC 1.1.1.157) claim 1 , 3-hydroxybutyryl-CoA dehydratase (Crt claim 1 , EC 4.2.1.5) claim 1 , trans-enoyl-CoA reductase (Ter claim 1 , EC 1.3.1.38) and aldehyde/alcohol dehydrogenase (AdhE2 claim 1 , EC 1.2.157/EC 1.1.1.1).4. The system of wherein in the engineered cell claim 1 , the endogenous fumarate reductase (FrdBC claim 1 , EC 1.3.1.6) claim 1 , lactate dehydrogenase (Ldh claim 1 , EC 1.1.1.27) claim 1 , acetaldehyde dehydrogenase (AdhE claim 1 , EC 1.2.1.10) ...

Methods And Microorganisms For The Production Of 1,3-Butanediol

A non-naturally occurring microorganism having a 1,3-BDO pathway is provided. The microorganism expresses at least one of the following 1,3-BDO pathway enzymes: an aldolase that catalyzes condensation of two acetaldehydes to produce 3-hydroxybutanal; and an aldo-ketoreductase, oxidoreductase, aldehyde reductase or alcohol dehydrogenase that reduces 3-hydroxybutanal to 1,3-BDO. The organism may further express one or more enzymes for producing acetaldehyde. A biosynthetic process involves condensing two acetaldehyde molecules to 3-hydroxybutanal using an enzyme from class aldolases; and selectively reducing 3-hydroxybutanal to 1,3-BDO using an enzyme belonging to the class aldo-ketoreductase, oxidoreductase, aldehyde reductase or alcohol dehydrogenase. The process can further include producing acetaldehyde by a biosynthetic method. 125-. (canceled)26. A non-naturally occurring microorganism that produces 1 ,3-butanediol (1 ,3-BDO) from 3-hydroxybutanal , the microorganism being engineered to express an exogenous polynucleotide encoding an aldo-keto reductase (AKR) comprising an amino acid sequence at least 85% identical to SEQ ID NO: 25 , wherein the AKR catalyzes reduction of the 3-hydroxybutanal to 1 ,3-BDO.27. The non-naturally occurring microorganism of claim 26 , wherein the AKR comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 25.28. The non-naturally occurring microorganism of claim 26 , wherein the AKR comprises residues R214 claim 26 , R227 claim 26 , R281 claim 26 , Q285 claim 26 , G279 claim 26 , and R208 claim 26 , with respect to the amino acid numbering of SEQ ID NO: 25.29. The non-naturally occurring microorganism of claim 26 , which is further engineered to express at least one further exogenous polynucleotide encoding a decarboxylase that catalyzes the decarboxylation of pyruvate to yield acetaldehyde and carbon dioxide.30. The non-naturally occurring microorganism of claim 29 , wherein the decarboxylase ...

Improved Carbon Dioxide Fixation Via Bypassing Feedback Regulation

Genetically engineered cells and methods are presented that allow for the production of various value products from CO. Contemplated cells have a CBB cycle that is genetically modified such that two molecules of COfixed in the CBB cycle can be withdrawn from the modified CBB cycle as a single C2 compound. In contemplated aspects a CBB cycle includes an enzymatic activity that generates the single C2 compound from a compound of the CBB cycle, while further modifications to the CBB cycle will not introduce additional recombinant enzymatic activity/activities outside the already existing catalytic activities in the CBB cycle. 1. A method for improving the efficiency of carbon dioxide fixation in an organism having a CBB cycle , comprising:genetically modifying the organism with the CBB cycle to produce or overexpress a first enzyme with a phosphoketolase activity, and to produce or overexpress a second enzyme with a phosphoribulokinase activity;wherein the first enzyme utilizes an intermediate of the CBB pathway as a substrate and generates a first acetyl phosphate product;wherein the phosphoribulokinase activity is produced or overexpressed in an amount to achieve a phosphoribulokinase activity level that is higher than the native phosphoribulokinase activity level of the organism; andwherein the first acetyl phosphate product is converted in the organism to acetyl-CoA.2. The method of claim 1 , wherein the intermediate of the CBB pathway is fructose-6-phosphate.3. The method of claim 1 , wherein the genetically modified organism fixes COin a medium containing nitrogen in an amount of less than 3 mM.4. The method of claim 1 , wherein the genetically modified organism is further modified to produce from the acetyl-CoA a value added product selected from the group consisting of an alcohol claim 1 , a fuel claim 1 , a plastic polymer claim 1 , and a monomer suitable for plastic polymer synthesis.5. The method of claim 1 , wherein the genetically modified organism is ...

Self-Assembled Nanoplatelet-Enzyme Bioconjugates Providing for Increased Biocatalytic Efficiency

A nanoplatelet serves as a substrate for immobilizing enzymes involved in consecutive reactions as a cascade. This results in a significant increase in the rate of catalysis as well as final product yield compared to non-immobilized enzymes or enzymes immobilized to quantum dots.

Methods And Microorganisms For The Production Of 1,3-Butanediol

A non-naturally occurring microorganism having a 1,3-BDO pathway is provided. The microorganism expresses at least one of the following 1,3-BDO pathway enzymes: an aldolase that catalyzes condensation of two acetaldehydes to produce 3-hydroxybutanal; and an aldo-ketoreductase, oxidoreductase, aldehyde reductase or alcohol dehydrogenase that reduces 3-hydroxybutanal to 1,3-BDO. The organism may further express one or more enzymes for producing acetaldehyde. A biosynthetic process involves condensing two acetaldehyde molecules to 3-hydroxybutanal using an enzyme from class aldolases; and selectively reducing 3-hydroxybutanal to 1,3-BDO using an enzyme belonging to the class aldo-ketoreductase, oxidoreductase, aldehyde reductase or alcohol dehydrogenase. The process can further include producing acetaldehyde by a biosynthetic method. 1. A non-naturally occurring microorganism having a 1 ,3-BDO pathway , wherein the microorganism comprises at least one of the following 1 ,3-BDO pathway enzymes:an aldolase that catalyzes condensation of two acetaldehydes to produce 3-hydroxybutanal; andan aldo-ketoreductase, oxidoreductase, aldehyde reductase or alcohol dehydrogenase that reduces 3-hydroxybutanal to 1,3-BDO;wherein the microorganism comprises at least one exogenous nucleic acid encoding an enzyme from said 1,3-BDO pathway.2. The non-naturally occurring microorganism according to claim 1 , wherein the microorganism comprises at least one modification to an endogenous nucleic acid encoding an enzyme from said 1 claim 1 ,3-BDO pathway or affecting the expression of an enzyme from said 1 claim 1 ,3-BDO pathway.3. The non-naturally occurring microorganism according to claim 1 , wherein the microorganism comprises an exogenous nucleic acid that encodes the aldolase.4. The non-naturally occurring microorganism according to claim 1 , wherein the aldolase is deoxyribose-5-phosphate aldolase (DERA).5. The non-naturally occurring microorganism according to claim 4 , wherein the ...

REDOX BALANCING IN YEAST

Described are composition and methods relate to reducing the redox imbalance in anaerobically growing yeast with attenuated glycerol production by re-engineering the pathway for Ac-CoA biosynthesis. 1. Modified yeast cells , comprising: an attenuated native biosynthetic pathway for making Ac-CoA , which native pathway contributes to redox cofactor imbalance in the cells under anaerobic conditions; introduction of an artificial alternative pathway for making Ac-CoA , which artificial pathway does not contribute to a redox cofactor imbalance in the cells under anaerobic conditions compared to the native biosynthetic pathway; and attenuation of the glycerol biosynthesis pathway; wherein the modified yeast cells demonstrate increased ethanol production using a carbohydrate substrate compared to a comparable yeast cells lacking the modifications.2. The modified yeast cells of claim 1 , wherein attenuation of the native Ac-CoA pathway is achieved by reducing aldehyde dehydrogenase activity.3. The modified yeast of or claim 1 , wherein attenuation of the native Ac-CoA pathway is achieved by reducing the expression of one or more of the native genes encoding aldehyde dehydrogenase (ALD2 claim 1 , ALD3 claim 1 , ALD4 claim 1 , ALD5 or ALD6).4. The modified yeast cells of any of - claim 1 , wherein the artificial alternative pathway for making Ac-CoA is the result of introducing exogenous phosphoketolase activity and exogenous phosphotransacetylase activity.5. The modified yeast cells of any of - claim 1 , wherein the artificial alternative pathway for making Ac-CoA is the result of introducing a heterologous phosphoketolase gene and a heterologous phosphotransacetylase gene.6. The modified yeast cells of any of - claim 1 , wherein attenuation of the glycerol biosynthesis pathway is the disruption or modification of GDP1 claim 1 , GDP2 claim 1 , GPP1 and/or GPP2.7. The modified yeast cells of any of the preceding claims claim 1 , wherein the cells further comprise increased ...

RECOMBINANT CELL, AND METHOD FOR PRODUCING 1,4-BUTANEDIOL

An object of the present invention is to provide a series of techniques for producing 1,4-butanediol from methanol or the like. Provided is a recombinant cell prepared by introducing a gene encoding at least one enzyme selected from the group consisting of succinate semialdehyde dehydrogenase, succinyl-CoA synthase, CoA-dependent succinate semialdehyde dehydrogenase, 4-hydroxybutyrate dehydrogenase, 4-hydroxybutyryl-CoA transferase, 4-hydroxybutyryl-CoA reductase, 4-hydroxybutyraldehyde dehydrogenase, and alcohol dehydrogenase, into a host cell which is a methylotroph, wherein the gene is expressed in the host cell, and the recombinant cell is capable of producing 1,4-butanediol from at least one C1 compound selected from the group consisting of methane, methanol, methylamine, formic acid, formaldehyde, and formamide. 121-. (canceled)23. The recombinant cell according to claim 22 , having at least one C1 carbon assimilating pathway selected from the group consisting of a serine pathway claim 22 , a ribulose monophosphate pathway claim 22 , and a xylulose monophosphate pathway as a fixing pathway of formaldehyde.24. The recombinant cell according to claim 22 , wherein a gene encoding 3-hexulose-6-phosphate synthase and a gene encoding 6-phospho-3-hexuloisomerase are further introduced claim 22 , and the genes are expressed in the host cell.25. The recombinant cell according to claim 22 , wherein the recombinant cell is tolerant to at least 400 mM 1 claim 22 ,4-butanediol.26. The recombinant cell according to claim 22 , wherein the recombinant cell is tolerant to at least 2% (v/v) methanol.28. The recombinant cell according to claim 27 , wherein the gene imparting formaldehyde fixing ability is a gene encoding 3-hexulose 6 phosphate synthase and a gene encoding 6-phospho-3-hexuloisomerase.29. The recombinant cell according to claim 27 , having at least one C1 carbon assimilating pathway selected from the group consisting of a serine pathway claim 27 , a ribulose ...

MODIFIED MICROORGANISMS AND METHODS FOR PRODUCTION OF USEFUL PRODUCTS

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

METHODS FOR ENHANCING MICROBIAL PRODUCTION OF SPECIFIC LENGTH FATTY ALCOHOLS IN THE PRESENCE OF METHANOL

The invention provides non-naturally occurring microbial organisms having a formaldehyde fixation pathway, a formate assimilation pathway, and/or a methanol metabolic pathway in combination with a fatty alcohol, fatty aldehyde, fatty acid or isopropanol pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length or isopropanol. The microbial organisms provided advantageously enhance the production of substrates and/or pathway intermediates for the production of chain length specific fatty alcohols, fatty aldehydes, fatty acids or isopropanol. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde, a fatty acid or isopropanol. 2. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism has a formaldehyde fixation pathway and a MI-FAE cycle in combination with a termination pathway.3. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism has a formate assimilation pathway and a MI-FAE cycle in combination with a termination pathway.4. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism has a formaldehyde fixation pathway claim 1 , a formate assimilation pathway claim 1 , and a MI-FAE cycle in combination with a termination pathway.5. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism has a formaldehyde fixation pathway and a MD-FAE cycle in combination with a termination pathway.6. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism has a formate assimilation pathway and a MD-FAE cycle in combination with a termination pathway.7. The non-naturally ...

METHOD FOR PRODUCING CURED TOBACCO MATERIAL

Provided is a method of producing a cured tobacco material having an enhanced threonine content. The present invention provides a method of producing a cured tobacco material having an enhanced threonine content, wherein the cured tobacco material is produced from a modified tobacco plant which has been modified such that expression or activity of an endogenous threonine aldolase is reduced or from a part of the modified tobacco plant. 1. A method of producing a cured tobacco material having an enhanced threonine content ,comprising producing the cured tobacco material from (i) a modified tobacco plant which has been modified such that expression or activity of an endogenous threonine aldolase is reduced or (ii) a part of the modified tobacco plant,said endogenous threonine aldolase being at least one of the following threonine aldolases (a) and (b):(a) a threonine aldolase comprised of a polypeptide encoded by a gene containing a polynucleotide which has a sequence identity of not less than 90% to a polynucleotide which has a base sequence shown in SEQ ID NO:1 or a threonine aldolase comprised of a polypeptide which has a sequence identity of not less than 90% to a polypeptide which has an amino acid sequence shown in SEQ ID NO:2; and(b) a threonine aldolase comprised of a polypeptide encoded by a gene containing a polynucleotide which has a sequence identity of not less than 90% to a polynucleotide which has a base sequence shown in SEQ ID NO:3 or a threonine aldolase comprised of a polypeptide which has a sequence identity of not less than 90% to a polypeptide which has an amino acid sequence shown in SEQ ID NO:4.2. (canceled)3. The method according to claim 1 , wherein a reduction in expression or in activity of the endogenous threonine aldolase in the modified tobacco plant is caused by (i) a mutant allele of a gene coding for the endogenous threonine aldolase or (ii) an exogenous polynucleotide which has been introduced into the modified tobacco plant in order ...