Microorganisms and methods for the coproduction 1,4-butanediol and gamma-butyrolactone

Patent metadata

Field	Value
Publication number	US-9988656-B2
Application number	US-201514954487-A
Country	US
Kind code	B2
Filing date	Nov 30, 2015
Priority date	Nov 25, 2009
Publication date	Jun 5, 2018
Grant date	Jun 5, 2018

Abstract

The invention provides non-naturally occurring microbial organisms comprising 1,4-butanediol (14-BDO) and gamma-butyrolactone (GBL) pathways comprising at least one exogenous nucleic acid encoding a 14-BDO and GBL pathway enzyme expressed in a sufficient amount to produce 14-BDO and GBL. The invention additionally provides methods of using such microbial organisms to produce 14-BDO and GBL.

First claim

What is claimed is:

1. A non-naturally occurring microbial organism comprising exogenous nucleic acids encoding a combination of enzymes of:
a 4-hydroxybutyrate dehydrogenase; a 4-hydroxybutyryl-CoA transferase; a 4-hydroxybutyryl-CoA reductase (aldehyde forming); a 1,4-butanediol dehydrogenase; and a 4-hydroxybutyrate lactonase;
wherein the exogenous nucleic acids are expressed in the microbial organism.

2. The non-naturally occurring microbial organism of claim 1 , wherein the microbial organism further comprises exogenous nucleic acids encoding a second combination of enzymes selective from the group consisting of:
(a) a succinyl-CoA transferase, a succinyl-CoA hydrolase, or a succinyl-CoA synthetase; and a succinyl-CoA reductase (aldehyde forming);
(b) an alpha-ketoglutarate decarboxylase; and
(c) a glutamate dehydrogenase or glutamate transaminase; a glutamate decarboxylase; and a 4-aminobutyrate dehydrogenase or a 4-aminobutyrate transaminase,
wherein the exogenous nucleic acids encoding the second combination of enzymes are expressed in the microbial organism.

3. The non-naturally occurring microbial organism of claim 1 , wherein at least one, two, three, four or five of the exogenous nucleic acids is a heterologous encoding nucleic acid.

4. The non-naturally occurring microbial organism of claim 1 , wherein the exogenous nucleic acids are heterologous encoding nucleic acids.

5. The non-naturally occurring microbial organism of claim 1 , wherein the microbial organism is a bacterium, yeast or fungus.

6. The non-naturally occurring microbial organism of claim 5 , wherein the bacterium is selected from the group consisting of Escherichia coli, Klebsiella oxytoca, Anaerobiospirillum succiniciproducens, Actinobacillus succinogenes, Mannheimia succiniciproducens, Rhizobium etli, Bacillus subtilis, Corynebacterium glutamicum, Gluconobacter oxydans, Zymomonas mobilis, Lactococcus lactis, Lactobacillus plantarum, Streptomyces coelicolor, Clostridium acetobutylicum, Pseudomonas fluorescens , and Pseudomonas putida.

7. The non-naturally occurring microbial organism of claim 5 , wherein the yeast or fungus is selected from the group consisting of Schizosaccharomyces pombe, Kluyveromyces lactis, Kluyveromyces marxianus, Aspergillus terreus, Aspergillus niger, Pichia pastoris, Rhizopus arrhizus , and Rhizobus oryzae.

8. A composition comprising the non-naturally occurring microbial organism of claim 1 in a substantially anaerobic culture medium.

Assignees

• Genomatica Inc

Inventors

• Sun Jun
• Burk Mark J
• Burgard Anthony P
• Osterhout Robin E
• Niu Wei
• Trawick John D
• Haselbeck Robert

Classifications

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12P7/18

  polyhydric · CPC title

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• C12P17/04 Primary

  containing a five-membered hetero ring, e.g. griseofulvin {, vitamin C} · CPC title