Microorganisms and methods for the biosynthesis of butadiene

Patent metadata

Field	Value
Publication number	US-9732361-B2
Application number	US-201314059131-A
Country	US
Kind code	B2
Filing date	Oct 21, 2013
Priority date	May 5, 2010
Publication date	Aug 15, 2017
Grant date	Aug 15, 2017

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 non-naturally occurring microbial organism, said microbial organism having a butadiene pathway and comprising at least one exogenous nucleic acid encoding a butadiene pathway enzyme expressed in a sufficient amount to produce butadiene, said butadiene pathway comprising converting crotonyl-CoA to crotyl-alcohol using one or more butadiene pathway enzymes, wherein said butadiene pathway enzymes are selected from the group consisting of a crotonyl-CoA reductase (aldehyde forming), a crotonaldehyde reductase (alcohol forming), a crotonyl-CoA hydrolase, synthetase, or transferase, a crotonate reductase, and a crotonyl-CoA reductase (alcohol forming).

2. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises two exogenous nucleic acids each encoding a butadiene pathway enzyme.

3. The non-naturally occurring microbial organism of claim 1 , wherein said butadiene pathway enzymes comprise a crotonyl-CoA reductase (aldehyde forming), and a crotonaldehyde reductase (alcohol forming).

4. The non-naturally occurring microbial organism of claim 1 , wherein said butadiene pathway enzymes comprise a crotonyl-CoA reductase (alcohol forming).

5. The non-naturally occurring microbial organism of claim 1 , wherein said butadiene pathway enzymes comprise a crotonyl-CoA hydrolase, synthetase or transferase and a crotonate reductase.

6. The non-naturally occurring microbial organism of claim 1 , wherein said butadiene pathway further comprises converting crotyl alcohol to butadiene using butadiene pathway enzymes, wherein said butadiene pathway enzymes are a crotyl alcohol kinase, a 2-butenyl-4-phosphate kinase and a butadiene synthase.

7. The non-naturally occurring microbial organism of claim 1 , wherein said butadiene pathway further comprises converting crotyl alcohol to butadiene using butadiene pathway enzymes, wherein said butadiene pathway enzymes are a butadiene synthase and a crotyl alcohol diphosphokinase.

8. The non-naturally occurring microbial organism of claim 3 , wherein said butadiene pathway further comprises converting acetyl-CoA to crotonyl-CoA using an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase and a 3-hydroxybutyryl-CoA dehydratase.

9. The non-naturally occurring microbial organism of claim 4 , wherein said butadiene pathway further comprises converting acetyl-CoA to crotonyl-CoA using an acetyl-CoA:acetyl-CoA acyltransferase, an acetoacetyl-CoA reductase and a 3-hydroxybutyryl-CoA dehydratase.

10. The non-naturally occurring microbial organism of claim 1 , wherein said butadiene pathway further comprises converting glutaconyl-CoA to crotonyl-CoA using a glutaconyl-CoA decarboxylase.

11. The non-naturally occurring microbial organism of claim 1 , wherein said butadiene pathway further comprises converting glutaryl-CoA to crotonyl-CoA using a glutaryl-CoA dehydrogenase.

12. The non-naturally occurring microbial organism of claim 1 , wherein said butadiene pathway further comprises converting 3-aminobutyryl-CoA to crotonyl-CoA using an 3-aminobutyryl-CoA deaminase.

13. The non-naturally occurring microbial organism of claim 1 , wherein said butadiene pathway further comprises converting 4-hydroxybutyryl-CoA to crotonyl-CoA using a 4-hydroxybutyryl-CoA dehydratase.

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

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

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

17. The method of claim 16 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.

18. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism is a bacteria, a yeast, or a fungus.

19. The non-naturally occurring microbial organism of claim 18 , wherein said a bacteria is Escherichia coli.

20. The non-naturally occurring microbial organism of claim 18 , wherein said a yeast is Saccharomyces cerevisiae.

Assignees

• Genomatica Inc

Inventors

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

Classifications

• C12P7/04

  acyclic · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12N1/18

  Baker's yeast; Brewer's yeast · CPC title

• C12P5/026 Primary

  Unsaturated compounds, i.e. alkenes, alkynes or allenes · CPC title

• C12N1/14

  Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor · CPC title