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

Patent metadata

Field	Value
Publication number	US-10513716-B2
Application number	US-201314053381-A
Country	US
Kind code	B2
Filing date	Oct 14, 2013
Priority date	Oct 15, 2012
Publication date	Dec 24, 2019
Grant date	Dec 24, 2019

Abstract

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.

First claim

What is claimed is:

1. A non-naturally occurring microbial organism for production of a compound of Formula (VI), (X) or (XIV):

wherein at least 80% of the compound produced by the microbial organism is C 7 linear alkyl at R 1 ,
wherein the microbial organism comprises a malonyl-CoA independent fatty acyl-CoA elongation (MI-FAE) cycle and a termination pathway,
wherein said MI-FAE cycle comprises one or more thiolase, one or more 3-oxoacyl-CoA reductase, one or more 3-hydroxyacyl-CoA dehydratase, and one or more enoyl-CoA reductase,
wherein said termination pathway comprises a pathway selected from:
(1) 1H;
(2) 1K and 1L;
(3) 1E and 1N;
(4) 1K, 1J, and 1N;
(5) 1E;
(6) 1K and 1J;
(7) 1H and 1N;
(8) 1K, 1L, and 1N;
(9) 1E and 1F;
(10) 1K, 1J, and 1F;
(11) 1H, 1N, and 1F;
(12) 1K, 1L, 1N, and 1F; and
(13) 1G,

wherein 1E is an acyl-CoA reductase (aldehyde forming), wherein 1F is an alcohol dehydrogenase, wherein 1G is an acyl-CoA reductase (alcohol forming), wherein 1H is an acyl-CoA hydrolase, acyl-CoA transferase or acyl-CoA synthase, wherein 1J is an acyl-ACP reductase, wherein 1K is an acyl-CoA:ACP acyltransferase, wherein 1L is a thioesterase, wherein 1N is an aldehyde dehydrogenase (acid forming) or a carboxylic acid reductase,
wherein an enzyme of the MI-FAE cycle or termination pathway is encoded by at least one exogenous nucleic acid and is expressed in a sufficient amount to produce the compound of Formula (VI), (X) or (XIV),
wherein one or more enzymes of the MI-FAE cycle are each independently selective for a compound of Formula (II), propionyl-CoA or acetyl-CoA as a substrate:

wherein R 1a is no greater than C 7 linear alkyl R 3 is H, OH, or oxo (═O); R 4 is S-CoA; and represents a single or double bond with the proviso that the valency of the carbon atom to which R 3 is attached is four; and
wherein one or more enzymes of the termination pathway are each independently selective for the compound of Formula (II) as a substrate wherein: R 1a is no less than C 7 linear alkyl; R 3 is H; and R 4 is S-CoA, ACP, OH or H.

2. A non-naturally occurring microbial organism for production of a compound of Formula (VI), (X) or (XIV):

wherein at least 80% of the compound produced by the microbial organism is C 9 linear alkyl at R 1 ,
wherein the microbial organism comprises a malonyl-CoA independent fatty acyl-CoA elongation (MI-FAE) cycle and a termination pathway,
wherein said MI-FAE cycle comprises one or more thiolase, one or more 3-oxoacyl-CoA reductase, one or more 3-hydroxyacyl-CoA dehydratase, and one or more enoyl-CoA reductase,
wherein said termination pathway comprises a pathway selected from:
(1) 1H;
(2) 1K and 1L;
(3) 1E and 1N;
(4) 1K, 1J, and 1N;
(5) 1E;
(6) 1K and 1J;
(7) 1H and 1N;
(8) 1K, 1L, and 1N;
(9) 1E and 1F;
(10) 1K, 1J, and 1F;
(11) 1H, 1N, and 1F;
(12) 1K, 1L, 1N, and 1F; and
(13) 1G,
wherein 1E is an acyl-CoA reductase (aldehyde forming), wherein 1F is an alcohol dehydrogenase, wherein 1G is an acyl-CoA reductase (alcohol forming), wherein 1H is an acyl-CoA hydrolase, acyl-CoA transferase or acyl-CoA synthase, wherein 1J is an acyl-ACP reductase, wherein 1K is an acyl-CoA:ACP acyltransferase, wherein 1L is a thioesterase, wherein 1N is an aldehyde dehydrogenase (acid forming) or a carboxylic acid reductase,
wherein an enzyme of the MI-FAE cycle or termination pathway is encoded by at least one exogenous nucleic acid and is expressed in a sufficient amount to produce the compound of Formula (VI), (X) or (XIV),
wherein one or more enzymes of the MI-FAE cycle are each independently selective for a compound of Formula (II), propionyl-CoA or acetyl-CoA as a substrate:

wherein R 1a is no greater than C 9 linear alkyl; R 3 is H, OH, or oxo (═O); R 4 is S-CoA; and represents a single or double bond with the proviso that the valency of the carbon atom to which R 3 is attached is four; and
wherein one or more enzymes of the termination pathway are each independently selective for the compound of Formula (II) as a substrate wherein: R 1a is no less than C 9 linear alkyl; R 3 is H; and R 4 is S-CoA, ACP, OH or H.

3. A non-naturally occurring microbial organism for production of a compound of Formula (VI), (X) or (XIV):

wherein at least 80% of the compound produced by the microbial organism is C 11 linear alkyl at R 1 ,
wherein the microbial organism comprises a malonyl-CoA independent fatty acyl-CoA elongation (MI-FAE) cycle and a termination pathway,
wherein said MI-FAE cycle comprises one or more thiolase, one or more 3-oxoacyl-CoA reductase, one or more 3-hydroxyacyl-CoA dehydratase, and one or more enoyl-CoA reductase,
wherein said termination pathway comprises a pathway selected from:
(1) 1H;
(2) 1K and 1L;
(3) 1E and 1N;
(4) 1K, 1J, and 1N;
(5) 1E;
(6) 1K and 1J;
(7) 1H and 1N;
(8) 1K, 1L, and 1N;
(9) 1E and 1F;
(10) 1K, 1J, and 1F;
(11) 1H, 1N, and 1F;
(12) 1K, 1L, 1N, and 1F; and
(13) 1G,
wherein 1E is an acyl-CoA reductase (aldehyde forming), wherein 1F is an alcohol dehydrogenase, wherein 1G is an acyl-CoA reductase (alcohol forming), wherein 1H is an acyl-CoA hydrolase, acyl-CoA transferase or acyl-CoA synthase, wherein 1J is an acyl-ACP reductase, wherein 1K is an acyl-CoA:ACP acyltransferase, wherein 1L is a thioesterase,
wherein 1N is an aldehyde dehydrogenase (acid forming) or a carboxylic acid reductase, wherein an enzyme of the MI-FAE cycle or termination pathway is encoded by at least one exogenous nucleic acid and is expressed in a sufficient amount to produce the compound of Formula (VI), (X) or (XIV),
wherein one or more enzymes of the MI-FAE cycle are each independently selective for a compound of Formula (II), propionyl-CoA or acetyl-CoA as a substrate:

wherein R 1a is no greater than C 11 linear alkyl; R 3 is H, OH, or oxo (═O); R 4 is S-CoA; and represents a single or double bond with the proviso that the valency of the carbon atom to which R 3 is attached is four; and
wherein one or more enzymes of the termination pathway are each independently selective for the compound of Formula (II) as a substrate wherein: R 1a is no less than C 11 linear alkyl; R 3 is H; and R 4 is S-CoA, ACP, OH or H.

4. A non-naturally occurring microbial organism for production of a compound of Formula (VI), (X) or (XIV):

wherein at least 80% of the compound produced by the microbial organism is C 13 linear alkyl at R 1 ,
wherein the microbial organism comprises a malonyl-CoA independent fatty acyl-CoA elongation (MI-FAE) cycle and a termination pathway,
wherein s

Assignees

• Genomatica Inc

Inventors

• Osterhout Robin E
• Burgard Anthony P

Classifications

• C07C29/00

  Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring · CPC title

• C12N9/0008

  acting on the aldehyde or oxo group of donors (1.2) · CPC title

• C12Y102/01076

  Succinate-semialdehyde dehydrogenase (acetylating) (1.2.1.76) · CPC title

• C12P7/6409 Primary

  Fatty acids · CPC title

• C12P7/04 Primary

  acyclic · CPC title