Microorganisms and methods for the biosynthesis of butadiene

Patent metadata

Field	Value
Publication number	US-9321701-B2
Application number	US-201213365132-A
Country	US
Kind code	B2
Filing date	Feb 2, 2012
Priority date	Feb 2, 2011
Publication date	Apr 26, 2016
Grant date	Apr 26, 2016

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

What is claimed is:

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 thereby biosynthetically producing crotyl alcohol, wherein said microbial organism comprises a crotyl alcohol pathway comprising at least two exogenous nucleic acids encoding crotyl alcohol pathway enzymes expressed in a sufficient amount to produce crotyl alcohol, wherein said crotyl alcohol pathway comprises converting crotonyl-CoA to crotyl alcohol using at least one exogenous nucleic acid of said two exogenous nucleic acids, wherein said crotyl alcohol pathway enzymes are selected from the group consisting of:
(i) a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming);
(ii) a crotonyl-CoA reductase (alcohol forming); and
(iii) a crotonyl-CoA hydrolase, synthetase, or transferase, a crotonate reductase, and a crotonaldehyde reductase (alcohol forming), and
wherein said crotyl alcohol pathway further comprises at least one exogenous nucleic acid of said two exogenous nucleic acids that encodes a crotyl alcohol pathway enzyme for converting:
(i) acetyl-CoA to crotonyl-CoA using an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase and a 3-hydroxybutyryl-CoA dehydratase;
(ii) glutaconyl-CoA to crotonyl-CoA using a glutaconyl-CoA decarboxylase;
(iii) glutaryl-CoA to crotonyl-CoA using a glutaryl-CoA dehydrogenase;
(iv) 3-aminobutyryl-CoA to crotonyl-CoA using an 3-aminobutyryl-CoA deaminase; or
(v) 4-hydroxybutyryl-CoA to crotonyl-CoA using a 4-hydroxybutyryl-CoA dehydratase; and
(b) converting the crotyl alcohol produced by culturing said non-naturally occurring microbial organism in step (a) to butadiene using a chemical procedure.

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 microbial organism comprises two, three, four, five, or six exogenous nucleic acids each encoding a crotyl alcohol pathway enzyme.

4. The process of claim 1 , wherein said crotyl alcohol pathway comprises 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).

5. The process of claim 1 , wherein said at least one exogenous nucleic acid is a heterologous nucleic acid.

6. The process of claim 1 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.

7. The process of claim 1 , wherein said crotyl alcohol pathway comprises an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase, a 3-hydroxybutyryl-CoA dehydratase, and a crotonyl-CoA reductase (alcohol forming).

8. The process of claim 1 , wherein said crotyl alcohol pathway comprises 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.

9. The process of claim 1 , wherein said crotyl alcohol pathway comprises a glutaconyl-CoA decarboxylase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming).

10. The process of claim 1 , wherein said crotyl alcohol pathway comprises a glutaconyl-CoA decarboxylase and a crotonyl-CoA reductase (alcohol forming).

11. The process of claim 1 , wherein said crotyl alcohol pathway comprises a glutaconyl-CoA decarboxylase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase, or transferase, and a crotonate reductase.

12. The process of claim 1 , wherein said crotyl alcohol pathway comprises a glutaryl-CoA dehydrogenase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming).

13. The process of claim 1 , wherein said crotyl alcohol pathway comprises a glutaryl-CoA dehydrogenase and a crotonyl-CoA reductase (alcohol forming).

14. The process of claim 1 , wherein said crotyl alcohol pathway comprises a glutaryl-CoA dehydrogenase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase, or transferase, and a crotonate reductase.

15. The process of claim 1 , wherein said crotyl alcohol pathway comprises a 3-aminobutyryl-CoA deaminase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming).

16. The process of claim 1 , wherein said crotyl alcohol pathway comprises a 3-aminobutyryl-CoA deaminase and a crotonyl-CoA reductase (alcohol forming).

17. The process of claim 1 , wherein said crotyl alcohol pathway comprises a 3-aminobutyryl-CoA deaminase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase or transferase and a crotonate reductase.

18. The process of claim 1 , wherein said crotyl alcohol pathway comprises a 4-hydroxybutyryl-CoA dehydratase, a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming).

19. The process of claim 1 , wherein said crotyl alcohol pathway comprises a 4-hydroxybutyryl-CoA dehydratase and a crotonyl-CoA reductase (alcohol forming).

20. The process of claim 1 , wherein said crotyl alcohol pathway comprises a 4-hydroxybutyryl-CoA dehydratase, a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase or transferase, and a crotonate reductase.

Assignees

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

Inventors

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

Classifications

• C07C2521/04

  Alumina · CPC title

• C07C2523/28

  Molybdenum · CPC title

• C07C2527/167

  Phosphates or other compounds comprising the anion (PnO3n+1)(n+2)- · CPC title

• C07C2523/31

  combined with bismuth · CPC title

• C12P7/04

  acyclic · CPC title