Recombinant microorganism and methods of production thereof

Patent metadata

Field	Value
Publication number	US-9359611-B2
Application number	US-201414180423-A
Country	US
Kind code	B2
Filing date	Feb 14, 2014
Priority date	Oct 22, 2010
Publication date	Jun 7, 2016
Grant date	Jun 7, 2016

Abstract

The invention relates, inter alia, to novel genetically modified microorganisms capable of using CO to produce 1-butanol and/or a precursor thereof, novel methyltransferases and nucleic acids encoding same, methods for producing genetically modified microorganisms using said novel methyltransferases, and methods of producing 1-butanol and/or a precursor thereof by microbial fermentation.

First claim

We claim as our invention:

1. A bacterium comprising exogenous thiolase, exogenous 3-hydroxybutyryl-CoA dehydrogenase, exogenous crotonase/crotonyl-CoA hydratase, exogenous butyryl-CoA dehydrogenase, and exogenous electron transport flavoproteins A and B, wherein the bacterium is derived from a parental bacterium of Clostridium autoethanogenum and wherein the bacterium does not comprise an exogenous nucleic acid encoding a bifunctional butyraldehyde/butanol dehydrogenase.

2. The bacterium of claim 1 , wherein the bacterium produces butyryl-CoA.

3. The bacterium of claim 1 , wherein the bacterium further comprises exogenous phosphotransbutyrylase and exogenous butyrate kinase.

4. The bacterium of claim 3 , wherein the bacterium produces butyrate.

5. The bacterium of claim 3 , wherein the bacterium further comprises endogenous or exogenous ferredoxin dependent aldehyde oxidoreductase.

6. The bacterium of claim 5 , wherein the bacterium produces butyraldehyde.

7. The bacterium of claim 5 , wherein the bacterium further comprises exogenous butanol dehydrogenase.

8. The bacterium of claim 7 , wherein the bacterium produces 1-butanol.

9. The bacterium of claim 7 , wherein the bacterium produces at least 0.075 grams of 1-butanol per liter of fermentation broth.

10. The bacterium of claim 9 , wherein the bacterium produces about 0.075 to about 20 grams of 1-butanol per liter of fermentation broth.

11. The bacterium of claim 10 , wherein the bacterium produces about 0.15 to about 1.54 grams of 1-butanol per liter of fermentation broth.

12. The bacterium of claim 1 , wherein one or more of the thiolase, 3-hydroxybutyryl-CoA dehydrogenase, crotonase/crotonyl-CoA hydratase, and butyryl-CoA dehydrogenase are derived from Clostridium acetobutylicum.

13. The bacterium of claim 12 , wherein the thiolase is encoded by a nucleic acid sequence of SEQ ID NO: 1, the 3-hydroxybutyryl-CoA dehydrogenase is encoded by a nucleic acid sequence of SEQ ID NO: 2, the crotonase/crotonyl-CoA hydratase is encoded by a nucleic acid sequence of SEQ ID NO: 3, and/or the butyryl-CoA dehydrogenase is encoded by a nucleic acid sequence of SEQ ID NO: 4.

14. The bacterium of claim 1 , wherein the electron transport flavoproteins A and B are derived from Clostridium acetobutylicum.

15. The bacterium of claim 14 , wherein the electron transport flavoprotein A is encoded by a nucleic acid sequence of SEQ ID NO: 5 and the electron transport flavoprotein B is encoded by a nucleic acid sequence of SEQ ID NO: 6.

Assignees

• Lanzatech New Zealand Ltd

Inventors

• Koepke Michael
• Simpson Sean Dennis
• Liew Fungmin

Classifications

• C12N9/0006

  acting on CH-OH groups as donors (1.1) · CPC title

• C12N9/0008

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

• C12P7/16

  Butanols · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12N15/74 Primary

  Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora · CPC title