Microorganisms and methods for the biosynthesis of butadiene

Patent metadata

Field	Value
Publication number	US-2021147881-A1
Application number	US-202016912409-A
Country	US
Kind code	A1
Filing date	Jun 25, 2020
Priority date	Feb 2, 2011
Publication date	May 20, 2021
Grant date	—

Abstract

The invention provides non-naturally occurring microbial organisms having a butadiene pathway. The invention additionally provides methods of using such organisms to produce butadiene.

First claim

1 . A process for the production of butadiene comprising:
(a) culturing by fermentation in a sufficient amount of nutrients and media a non-naturally occurring microbial organism that produces crotyl alcohol; and (b) converting crotyl alcohol produced by culturing said non-naturally occurring microbial organism to butadiene.

2 . The process of claim 1 , wherein step (b) is performed by chemical dehydration in the presence of a catalyst.

3 . The process of claim 1 , wherein said non-naturally occurring microbial organism comprises a crotyl alcohol pathway comprising at least one exogenous nucleic acid encoding a crotyl alcohol pathway enzyme expressed in a sufficient amount to produce crotyl alcohol, said crotyl alcohol pathway comprising an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a crotonyl-CoA reductase (aldehyde forming), a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase, or transferase, a crotonate reductase, a crotonyl-CoA reductase (alcohol forming), a glutaconyl-CoA decarboxylase, a glutaryl-CoA dehydrogenase, a 3-aminobutyryl-CoA deaminase, or a 4-hydroxybutyryl-CoA dehydratase.

4 . The process of claim 3 , wherein said microbial organism comprises two, three or four exogenous nucleic acids each encoding a crotyl alcohol pathway enzyme.

5 - 6 . (canceled)

7 . The process of claim 3 , wherein said crotyl alcohol pathway comprises a pathway selected from the group consisting of:
an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming); an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, and a crotonyl-CoA reductase (alcohol forming); an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase or transferase and a crotonate reductase; a glutaconyl-CoA decarboxylase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming); a glutaconyl-CoA decarboxylase and a crotonyl-CoA reductase (alcohol forming); a glutaconyl-CoA decarboxylase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase, or transferase and a crotonate reductase; a glutaryl-CoA dehydrogenase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming); a glutaryl-CoA dehydrogenase and a crotonyl-CoA reductase (alcohol forming); a glutaryl-CoA dehydrogenase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase, or transferase and a crotonate reductase; a 3-aminobutyryl-CoA deaminase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming); a 3-aminobutyryl-CoA deaminase and a crotonyl-CoA reductase (alcohol forming); a 3-aminobutyryl-CoA deaminase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase or transferase and a crotonate reductase; a 4-hydroxybutyryl-CoA dehydratase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming); a 4-hydroxybutyryl-CoA dehydratase and a crotonyl-CoA reductase (alcohol forming); and a 4-hydroxybutyryl-CoA dehydratase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase or transferase and a crotonate reductase.

8 - 21 . (canceled)

22 . The process of claim 3 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.

23 . The process of claim 3 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.

24 . A non-naturally occurring microbial organism comprising a crotyl alcohol pathway, said crotyl alcohol pathway comprising at least one exogenous nucleic acid encoding a crotyl alcohol pathway enzyme expressed in a sufficient amount to produce crotyl alcohol, said crotyl alcohol pathway comprising an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a crotonyl-CoA reductase (aldehyde forming), a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase, or transferase, a crotonate reductase, a crotonyl-CoA reductase (alcohol forming), a glutaconyl-CoA decarboxylase, a glutaryl-CoA dehydrogenase, a 3-aminobutyryl-CoA deaminase, or a 4-hydroxybutyryl-CoA dehydratase.

25 . The microbial organism of claim 24 , wherein said microbial organism comprises two, three or four exogenous nucleic acids each encoding a crotyl alcohol pathway enzyme.

26 - 27 . (canceled)

28 . The microbial organism of claim 24 , wherein said crotyl alcohol pathway comprises a pathway selected from the group consisting of:
an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming); an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, and a crotonyl-CoA reductase (alcohol forming); an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase or transferase and a crotonate reductase; a glutaconyl-CoA decarboxylase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming); a glutaconyl-CoA decarboxylase and a crotonyl-CoA reductase (alcohol forming); a glutaconyl-CoA decarboxylase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase, or transferase and a crotonate reductase; a glutaryl-CoA dehydrogenase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming); a glutaryl-CoA dehydrogenase and a crotonyl-CoA reductase (alcohol forming); a glutaryl-CoA dehydrogenase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase, or transferase and a crotonate reductase; a 3-aminobutyryl-CoA deaminase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming); a 3-aminobutyryl-CoA deaminase and a crotonyl-CoA reductase (alcohol forming); a 3-aminobutyryl-CoA deaminase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase or transferase and a crotonate reductase; a 4-hydroxybutyryl-CoA dehydratase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming); a 4-hydroxybutyryl-CoA dehydratase and a crotonyl-CoA reductase (alcohol forming); and a 4-hydroxybutyryl-CoA dehydratase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase or transferase and a crotonate reductase.

29 - 42 . (canceled)

43 . The microbial organism of claim 24 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.

44 . The microbial organism of claim 24 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.

Assignees

• Genomatica Inc

Inventors

• Burk Mark J
• Burgard Anthony P
• Sun Jun
• Osterhout Robin E
• Pharkya Priti

Classifications

• C12N15/52 Primary

  Genes encoding for enzymes or proenzymes · CPC title

• C12P5/02 Primary

  acyclic {(C12P5/007 takes precedence)} · CPC title

• C07C1/24 Primary

  by elimination of water · CPC title

• C07C2523/31

  combined with bismuth · CPC title

• C07C2523/745

  Iron · CPC title