Methods and organisms for utilizing synthesis gas or other gaseous carbon sources and methanol

Patent metadata

Field	Value
Publication number	US-9885064-B2
Application number	US-201414185709-A
Country	US
Kind code	B2
Filing date	Feb 20, 2014
Priority date	Jan 22, 2008
Publication date	Feb 6, 2018
Grant date	Feb 6, 2018

Abstract

The invention provides a non-naturally occurring microbial organism having an acetyl-CoA pathway and the capability of utilizing syngas or syngas and methanol. In one embodiment, the invention provides a non-naturally occurring microorganism, comprising one or more exogenous proteins conferring to the microorganism a pathway to convert CO, CO 2 and/or H 2 to acetyl-coenzyme A (acetyl-CoA), methyl tetrahydrofolate (methyl-THF) or other desired products, wherein the microorganism lacks the ability to convert CO or CO 2 and H 2 to acetyl-CoA or methyl-THF in the absence of the one or more exogenous proteins. For example, the microbial organism can contain at least one exogenous nucleic acid encoding an enzyme or protein in an acetyl-CoA pathway. The microbial organism is capable of utilizing synthesis gases comprising CO, CO 2 and/or H 2 , alone or in combination with methanol, to produce acetyl-CoA. The invention additionally provides a method for producing acetyl-CoA, for example, by culturing an acetyl-CoA producing microbial organism, where the microbial organism expresses at least one exogenous nucleic acid encoding an acetyl-CoA pathway enzyme or protein in a sufficient amount to produce acetyl-CoA, under conditions and for a sufficient period of time to produce acetyl-CoA.

First claim

What is claimed is:

1. A non-naturally occurring microbial organism, comprising a microbial organism having an acetyl-coenzyme A (acetyl-CoA) pathway able to convert CO 2 , CO or H 2 , or a combination thereof, and methanol to acetyl-CoA, and comprising methanol-methyltransferase that catalyzes transfer of a methyl group from methanol to form methylated corrinoid protein, and acetyl-CoA synthase/carbon monoxide dehydrogenase that catalyzes reduction of carbon dioxide to carbon monoxide, and synthesis of acetyl-CoA from carbon monoxide, Coenzyme A, and a methyl group from a methylated corrinoid-iron-sulfur protein, wherein the methanol-methyltransferase and acetyl-CoA synthase/carbon monoxide dehydrogenase are encoded by exogenous nucleic acids expressed in sufficient amounts to produce acetyl-CoA.

2. The non-naturally occurring microbial organism of claim 1 , wherein the acetyl-CoA synthase/carbon monoxide dehydrogenase comprises an enzyme or protein selected from the group consisting of methyltetrahydrofolate: corrinoid protein methyltransferase, corrinoid iron-sulfur protein, nickel-protein assembly protein, ferredoxin, acetyl-CoA synthase, carbon monoxide dehydrogenase and nickel-protein assembly protein.

3. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises three exogenous nucleic acids encoding an acetyl-CoA pathway enzyme or protein, one exogenous nucleic acid encoding the methanol-methyltransferase and another exogenous nucleic acid encoding the acetyl-CoA synthase/carbon monoxide dehydrogenase.

4. The non-naturally occurring microbial organism of claim 1 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.

5. The non-naturally occurring microbial organism of claim 1 , wherein said non-naturally occurring microbial organism further comprises pyruvate ferredoxin oxidoreductase.

6. The non-naturally occurring microbial organism of claim 1 , wherein said non-naturally occurring microbial organism further comprises hydrogenase.

7. The non-naturally occurring microbial organism of claim 1 , wherein the methanol-methyltransferase is selected from the group consisting of Methanosarcina barkeri MtaB1, Methanosarcina barkeri MtaB2, Methanosarcina barkeri MtaB3, Methanosarcina acetivorans MtaB1, Methanosarcina acetivorans MtaB1, Methanosarcina acetivorans MtaB2, Methanosarcina acetivorans MtaB3, and Moorella thermoacetica MtaB.

8. The non-naturally occurring microbial organism of claim 1 , wherein the non-naturally occurring microbial organism is Escherichia coli and the methanol-methyltransferase and acetyl-CoA synthase/carbon monoxide dehydrogenase are encoded by exogenous nucleic acids heterologous to Escherichia coli.

9. A method for producing acetyl-CoA, comprising culturing the non-naturally occurring microbial organism of claim 1 , under conditions and for a sufficient period of time to produce acetyl-CoA.

10. The method of claim 9 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.

Assignees

• Genomatica Inc

Inventors

• Burk Mark J
• Schilling Christophe H
• Burgard Anthony P
• Trawick John D

Classifications

• C12N9/93

  Ligases (6) · CPC title

• C12N15/70

  Vectors or expression systems specially adapted for E. coli · CPC title

• C12N9/0036

  acting on NADH or NADPH (1.6) · CPC title

• C12N9/1007

  Methyltransferases (general) (2.1.1.) · CPC title

• C12P7/18 Primary

  polyhydric · CPC title