WO2012177983A4 - Microorganisms for producing ethylene glycol and methods related thereto - Google Patents
Microorganisms for producing ethylene glycol and methods related thereto Download PDFInfo
- Publication number
- WO2012177983A4 WO2012177983A4 PCT/US2012/043714 US2012043714W WO2012177983A4 WO 2012177983 A4 WO2012177983 A4 WO 2012177983A4 US 2012043714 W US2012043714 W US 2012043714W WO 2012177983 A4 WO2012177983 A4 WO 2012177983A4
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- reductase
- microbial organism
- ethylene glycol
- decarboxylase
- glycolaldehyde
- Prior art date
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/18—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic polyhydric
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/52—Genes encoding for enzymes or proenzymes
Abstract
The invention provides non-naturally occurring microbial organisms having an ethylene glycol pathway. The invention additionally provides methods of using such organisms to produce ethylene glycol. The invention provides non-naturally occurring microbial organisms containing ethylene glycol patways comprising at least one exogenous nucleic acid encoding an ethylene glycol pathway enzyme expressed in a sufficient amount to produce ethylene glycol. The invention additionally provides methods of using such microbial organisms to produce ethylene glycol, by culturing a non-naturally occurring microbial organism containing ethylene glycol pathways as described herein under conditions and for a sufficient period of time to produce ethylene glycol.
Claims
1. A non-natural ly occurring microbial organism, comprising a microbial organism having an ethylene glycol pathway comprising at least one exogenous nucleic acid encoding an ethylene glycol pathway enzyme expressed in a sufficient amount to produce ethylene glycol; said non-naturally occurring microbial organism further comprising;
(i) a reductive TCA pathway comprising at least one exogenous nucleic acid encoding a reductive TCA pathway enzyme, wherein said at least one exogenous nucleic acid is selected from an ATP-citrate lyase, a citrate lyase, a citryl-CoA synthetase, a citryl-CoA lyase, a fumarate reductase, and an alpha-ketoglutarate: ferredoxin oxidoreductase; (it) a reductive TCA pathway comprising at least one exogenous nucleic acid encoding a reductive TC A pathway enzyme, wherein said at least one exogenous nucleic acid is selected from a pyruvate:ferredoxin oxidoreductase, a phosphoenolpyruvate carboxylase, a phosphocnolpyruvatc carboxykinase. a CO dehydrogenase, and an ¾ hydrogenase; or
(iii) at least one exogenous nucleic acid encodes an enzyme selected from a CO dehydrogenase, an ¾ hydrogenase, and combinations thereof; wherein said ethylene glycol pathway comprises a pathway selected from:
(A) a serine aminotransferase or a serine oxidoreductase (deaminating); a hydroxypyruvate decarboxylase, and a glycolaldehyde reductase;
(B) a serine aminotransferase or a serine oxidoreductase (deaminating); a hydroxypyruvate reductase, and a glycerate decarboxylase;
(C) a serine decarboxylase; an cthanolamine aminotransferase or an ethanolamine oxidoreductase (deaminating), and a glycolaldehyde reductase;
(D) a hydroxypyruvate reductase; a hydroxypyruvate decarboxylase, and a glycolaldehyde reductase; (E) a glycerate decarboxylase;
(F) glycerate dehydrogenase, a hydroxypyruvate isomerasc. a hydroxypyruvate decarboxylase aiid a glycolaldehyde reductase;
148 (G) a glyoxylate carboligase, a hydroxypyruvate isomerasc, a hydroxypyruvate decarboxylase and a gl colaldehyde reductase;
(H) a glyoxylate reductase; a glycolyl-CoA transferase or a glycoiyl-CoA synthetase: a glycolyi-CoA reductase (aldehyde forming), and a glycolaldehyde reductase; (1) a glyoxylate reductase; a glycolate reductase, and a glycolaldehyde reductase;
(J) a glyoxylate reductase; a glycolyl-CoA transferase or a glycolyl-CoA synthetase, and a glycolyl-CoA reductase (alcohol forming);
(K) a glyoxylate reductase, a glycolate kinase, a phosphotransglycolylase, glycolyl- CoA reductase (aldehyde forming) and a glycolaldehyde reductase; (L) a glyoxylate reductase, a glycolate kinase, a phosphotransglycolylase and a gJycolyl-CoA reductase (alcohol forming);
(M) a glyoxylate reductase, glycolate kinase, a glycolylphosphate reductase and a glycolaldehyde reductase;
(N) a 3-phosphohydroxypyruvate decarboxylase, a 2-oxoethyl-phosphate reductase, and a 2-hydroxyethyl-phosphatc phosphatase or an ethylene glycol kinase; and
(0) a 3-phosphohydroxypyruvate decarboxylase, a 2-oxoethyl-phospbate
phosphatase or a glycolaldehyde kinase, and a glycolaldehyde reductase.
2. The non-naturally occurring microbial organism of claim 1. wherein the ethylene glycol pathway of (D), (E) and/or (F) further comprises a 3-phosphoglycerate phosphatase, a gtycerate kinase, a 2-phosphoglycerate phosphatase, a gtyccratc-2-kinase, or a
glyceraldehyde dehydrogenase.
3. The non-naturally occurring microbial organism of claim 1 or claim 2, wherein said microbial organism comprising (i) further comprises an exogenous nucleic acid encoding an enzyme selected from a pyruvate: erredoxin oxidoreductase, an aconitase, an isocitrate dehydrogenase, a succinyl-CoA synthetase, a succinyl-CoA transferase, a fumarase, a malate dehydrogenase, an acetate kinase, a phosphotransacetvlase, an acetyl-CoA synthetase, an \TAD(P)H:ferrcdoxin oxidoreductase, ferredoxin, and combinations thereof.
4. The non-natural ly occurring microbial organism of claim I or claim 2. wherein said microbial organism comprising (ii) further comprises an exogenous nucleic acid encoding an enzyme selected from an aconitase, an isocitrate dehydrogenase, a succinyl-CoA synthetase, a succinyl-CoA transferase, a fumarase, a malate dehydrogenase, and combinations thereof.
5. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises two, three, four, five, six or seven exogenous nucleic acids each encoding a ethylene glycol pathway enzyme.
6. The non-naturally occurring microbial organism of claim 5, wherein said microbial organism comprises exogenous nucleic acids encoding each of the enzymes selected from:
(A) a serine aminotransferase or a serine oxidoreductase (deaminating); a hydToxypyruvate decarboxylase, and a glycolaldehyde reductase;
(B) a serine aminotransferase or a serine oxidoreductase (deaminating); a hydro pyruvate reductase, and a glycerate decarboxylase;
(C) a serine decarboxylase; an ethanolamine aminotransferase or an ethanolamine oxidoreductase (deaminating), and a glycolaldehyde reductase;
(D) a hydroxypyruvate reductase; a hydroxypyruvate decarboxylase, and a glycolaldehyde reductase; (E) a glycerate decarboxylase;
(F) glycerate dehydrogenase, a hydroxypyruvate isomerase, a hydroxypyruvate decarboxylase and a glycolaldehyde reductase;
(G) a glyoxylate carboligase, a hydroxypyruvate isomerase, a hydroxypyruvate decarboxylase and a glycolaldehyde reductase; (H) a glyoxylate reductase; a glycolyl-CoA transferase or a glycolyl-CoA synthetase; a glycolyl-CoA reductase (aldehyde forming), and a glycolaldehyde reductase;
(0 a glyoxylate reductase; a glycolate reductase, and a glycolaldehyde reductase; (J) a glyoxvlate reductase; a glycolyl-CoA transferase or a glycolyl-CoA synthetase, and a glycolyl-CoA reductase (alcohol forming);
( ) a glyoxylatc reductase, a glycolate kinase, a phosphotransglycolylase, glycolyl- CoA reductase (aldehyde forming) and a glycolaldehyde reductase; (L) a glyoxylatc reductase, a glycolate kinase, a phosphotransglycolylase and a glycolyl-CoA reductase (alcohol forming);
(M) a glyoxylale reductase, glycolate kinase, a glyco!ylphosphate reductase and a glycolaldehyde reductase;
(N) a 3-phosphohydroxypyruvate decarboxylase, a.2-oxoethyl-phosphate reductase, and a 2-hydroxyethyl-phosphate phosphatase or an ethylene glycol kinase; and
(0) a 3-phosphohydroxypyruvate decarboxylase, a 2-oxoethyl-phosphatc phosphatase or a glycolaldehyde kinase, and a glycolaldehyde reductase.
7. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises two, three, four or live exogenous nucleic acids each encoding enzymes of (i). (ii) or (iii).
8. The non-naturally occurring microbial organism of claim 7, wherein said microbial organism comprising (i) comprises three exogenous nucleic acids encoding an ATP-citrate lyase or a citrate lyase; a rumarate reductase; and an alpha-ketoglutarate:ferredoxin oxidorcductase; wherein said microbial organism comprising (ii) comprises four exogenous nucleic acids encoding a pyruvate:fen-edoxin oxidoreductase; a phosphoenolpyruvate carboxylase or a phosphoenolpyruvate carboxykinase; a CO dehydrogenase; and an ¾ hydrogenase; or wherein said microbial organism comprising (iii) comprises two exogenous nucleic acids encoding CO dehydrogenase and \ hydrogenase.
9. The non-naturally occurring microbial organism of claim 1, wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.
10. The non-natural !y occurring microbial organism of claim 1 , wherein said non- naturaliy occurring microbial organism is in a substantially anaerobic culture medium.
1 1. A non-naturally occurring microbial organism, said microbial organism having an ethylene glycol pathway and comprising at least one exogenous nucleic acid encoding an ethylene glycol pathway enzyme expressed in a sufficient amount to produce ethylene glycol, wherein said ethylene glycol pathway comprises a pathway selected from:
(A) a 3-phosphohydroxypyravate decarboxylase, a 2-oxoethyl-phosphate reductase, and a 2-hydroxycthyl-phosphate phosphatase or an eth lene glycol kinase; and
(B) a 3-phosphohydroxypymvate decarboxylase, a 2-oxoethyl-phosphate phosphatase or a glycolaldehydc kinase, and a glycolaldehyde reductase.
12. The non-naturally occurring microbial organism of claim 1 1. wherein said microbial organism comprises two or three exogenous nucleic acids each encoding an ethylene glycol pathway enzyme.
13. The non-naturally occurring microbial organism of claim 1 1. wherein said microbial organism comprises exogenous nucleic acids encoding each of the enzymes of at least one of the pathways selected from (A) and (B).
14. The non-naturally occurring microbial organism of claim 1 1 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.
15. The non-naturally occurring microbial organism of claim 1 1. wherein said non- naturally occurring microbial organism is in a substantially anaerobic culture medium.
16. A method for producing ethylene glycol, comprising culturing the non-naturally occurring microbial organism of any of claims 1 -15 under conditions and for a sufficient period of time to produce ethylene glycol.
152
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201161500106P | 2011-06-22 | 2011-06-22 | |
| US61/500,106 | 2011-06-22 | ||
| US201261620377P | 2012-04-04 | 2012-04-04 | |
| US61/620,377 | 2012-04-04 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| WO2012177983A2 WO2012177983A2 (en) | 2012-12-27 |
| WO2012177983A3 WO2012177983A3 (en) | 2013-05-16 |
| WO2012177983A4 true WO2012177983A4 (en) | 2013-06-20 |
Family
ID=47423227
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/US2012/043714 WO2012177983A2 (en) | 2011-06-22 | 2012-06-22 | Microorganisms for producing ethylene glycol and methods related thereto |
Country Status (1)
| Country | Link |
|---|---|
| WO (1) | WO2012177983A2 (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160040172A1 (en) | 2013-03-15 | 2016-02-11 | Genomatica, Inc. | Microorganisms and methods for producing butadiene and related compounds by formate assimilation |
| JP2016165225A (en) | 2013-07-09 | 2016-09-15 | 味の素株式会社 | Method for producing useful substance |
| SI3077501T1 (en) | 2013-12-03 | 2022-02-28 | Genomatica, Inc. | Microorganisms and methods for improving product yields on methanol using acetyl-coa synthesis |
| CA3192376A1 (en) | 2013-12-27 | 2015-07-02 | Genomatica, Inc. | Methods and organisms with increased carbon flux efficiencies |
| DK3194604T3 (en) | 2014-09-18 | 2020-05-25 | Genomatica Inc | NON-NATURAL MICROBIAL ORGANISMS WITH IMPROVED ENERGY EFFICIENCY |
| FR3028529B1 (en) | 2014-11-19 | 2016-12-30 | Inst Nat De La Rech Agronomique Inra | PROCESS FOR PRODUCING AT LEAST ONE METABOLITE OF INTEREST BY TRANSFORMING A PENTOSE IN A MICROORGANISM |
| US20210079334A1 (en) | 2018-01-30 | 2021-03-18 | Genomatica, Inc. | Fermentation systems and methods with substantially uniform volumetric uptake rate of a reactive gaseous component |
| US11384369B2 (en) | 2019-02-15 | 2022-07-12 | Braskem S.A. | Microorganisms and methods for the production of glycolic acid and glycine via reverse glyoxylate shunt |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI488964B (en) * | 2007-03-16 | 2015-06-21 | Genomatica Inc | Compositions and methods for the biosynthesis of 1,4-butanediol and its precursors |
| PT2379730E (en) * | 2008-12-31 | 2015-10-16 | Metabolic Explorer Sa | Method for the preparation of diols |
| EP2459711B1 (en) * | 2009-07-30 | 2016-04-06 | Metabolic Explorer | Mutant YqhD enzyme for the production of a biochemical by fermentation |
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2012
- 2012-06-22 WO PCT/US2012/043714 patent/WO2012177983A2/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| WO2012177983A3 (en) | 2013-05-16 |
| WO2012177983A2 (en) | 2012-12-27 |
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