Recombinant cell, and method for producing isoprene

Patent metadata

Field	Value
Publication number	US-9783828-B2
Application number	US-201314437034-A
Country	US
Kind code	B2
Filing date	Oct 22, 2013
Priority date	Oct 23, 2012
Publication date	Oct 10, 2017
Grant date	Oct 10, 2017

Abstract

To provide a series of techniques capable of producing isoprene from syngas or the like.
Provided is a recombinant cell prepared by introducing a nucleic acid encoding isoprene synthase into a host cell having an isopentenyl diphosphate synthesis ability by a non-mevalonate pathway, wherein the nucleic acid is expressed in the host cell, and the recombinant cell is capable of producing isoprene from at least one C1 compound selected from the group consisting of carbon monoxide, carbon dioxide, formic acid, and methanol. As the host cell, a Clostridium bacterium or a Moorella bacterium is exemplified. Also provided is a method for producing isoprene using the recombinant cell.

First claim

The invention claimed is:

1. A recombinant cell prepared by introducing a nucleic acid encoding isoprene synthase into a host cell wherein the host cell is a Clostridium bacterium or a Moorella bacterium, wherein the nucleic acid is expressed in the host cell, and the recombinant cell is capable of producing isoprene from at least one C1 compound selected from the group consisting of carbon monoxide and carbon dioxide, wherein a nucleic acid encoding a group of exogenous enzymes acting in a mevalonate pathway is further introduced so that an isopentenyl diphosphate synthesis ability by a mevalonate pathway is further imparted, and wherein the group of exogenous enzymes acting in a mevalonate pathway comprises mevalonate kinase, mevalonate diphosphate decarboxylase, phosphomevalonate kinase, isopentenyl diphosphate (IPP) isomerase, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, and HMG-CoA synthase.

2. The recombinant cell according to claim 1 , having carbon monoxide dehydrogenase.

3. The recombinant cell according to claim 1 , wherein the mevalonate pathway is that of yeast, prokaryote or actinomycete.

4. The recombinant cell according to claim 1 , wherein a nucleic acid encoding at least one enzyme acting in a non-mevalonate pathway is further introduced, and the nucleic acid is expressed in the host cell.

5. The recombinant cell according to claim 4 , wherein the non-mevalonate pathway is that of other organism than the host cell.

6. The recombinant cell according to claim 1 , wherein the isoprene synthase is derived from plant.

7. The recombinant cell according to claim 1 , wherein the nucleic acid encoding isoprene synthase encodes the following (a), (b) or (c):
(a) a protein consisting of an amino acid sequence of SEQ ID NO: 2,
(b) a protein consisting of an amino acid sequence in which 1 to 20 amino acids are deleted, substituted or added in the amino acid sequence of SEQ ID NO: 2, and having isoprene synthase activity, and
(c) a protein consisting of an amino acid sequence having a homology of 90% or more with the amino acid sequence of SEQ ID NO: 2, and having isoprene synthase activity.

8. The recombinant cell according to claim 1 , wherein the nucleic acid introduced into the host cell is codon-modified.

9. The recombinant cell according to claim 1 , wherein the nucleic acid introduced into the host cell is incorporated in a genome of the host cell.

10. The recombinant cell according to claim 1 , wherein the nucleic acid introduced into the host cell is incorporated in a plasmid.

11. The recombinant cell according to claim 1 , wherein the nucleic acid encoding isoprene synthase and the nucleic acid encoding a group of exogenous enzymes acting in a mevalonate pathway are regulated by a constitutive promoter.

12. A method for producing isoprene by culturing the recombinant cell according to claim 1 using at least one C1 compound selected from the group consisting of carbon monoxide and carbon dioxide as a carbon source, to allow the recombinant cell to produce isoprene.

13. A method for producing isoprene by bringing at least one C1 compound selected from the group consisting of carbon monoxide and carbon dioxide into contact with the recombinant cell according to claim 1 , to allow the recombinant cell to produce isoprene from the C1 compound.

14. The method according to claim 12 , wherein the recombinant cell is provided with a gas mainly containing carbon monoxide and hydrogen, or a gas mainly containing carbon dioxide and hydrogen.

15. The method according to claim 12 , wherein the recombinant cell is prepared from a Clostridium bacterium or a Moorella bacterium as a host cell, and isoprene released outside the recombinant cell is recovered.

16. The method according to claim 13 , wherein the recombinant cell is provided with a gas mainly containing carbon monoxide and hydrogen, or a gas mainly containing carbon dioxide and hydrogen.

17. The method according to claim 13 , wherein the recombinant cell is prepared from a Clostridium bacterium or a Moorella bacterium as a host cell, and isoprene released outside the recombinant cell is recovered.

Assignees

• Sekisui Chemical Co Ltd
• Fraunhofer-Gesellschaft Zur Forderung Der Angewandten Forschung E V

Inventors

• Furutani Masahiro
• Uenishi Akihiro
• Iwasa Koichiro
• Jennewein Stefan
• Fischer Rainer

Classifications

• C12N9/0008

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

• C12Y102/99002

  Carbon-monoxide dehydrogenase (acceptor) (1.2.99.2) · CPC title

• C12P5/007 Primary

  containing one or more isoprene units, i.e. terpenes (carotenes C12P23/00) · CPC title

• C12N9/88

  Lyases (4.) · CPC title

• C12Y302/01

  Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1) · CPC title