MICROORGANISM FOR PRODUCING PUTRESCINE OR ORNITHINE AND METHOD FOR PRODUCING PUTRESCINE OR ORNITHINE BY USING SAME

• US 20180208909A1
• Filed: 07/19/2016
• Published: 07/26/2018
• Est. Priority Date: 07/20/2015
• Status: Active Grant

• Current Assignee: CJ Cheiljedang Corporation (CJ Group)
• Original Assignee: CJ Cheiljedang Corporation (CJ Group)
• Inventors: PARK, Su Jin, YANG, Young Lyeol, UM, Hye Won, LI, Hong Xian, LEE, Kyoung Min, LEE, Baek Seok, LEE, Hyo Hyoung, JUNG, Hee Kyoung
• Granted Patent: US 10,640,753 B2
• Time in Patent Office: Days
• CPC Class Codes: C12N 1/00 Microorganisms, e.g. protozoa; Compositions thereof medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
• CPC Class Codes: C12N 15/00 Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor mutants or genetically engineered microorganisms, per se C12N1/00, C12N5/00, C12N7/00; new plants per se A01H; plant reproduction by tissue culture techniques A01H4/00; new animals per se A01K67/00; use of medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases, gene therapy A61K48/00
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 9/1018 Carboxy- and carbamoyl transferases (2.1.3)
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12N 9/1217 Phosphotransferases with a carboxyl group as acceptor (2.7.2)
• CPC Class Codes: C12N 9/80 acting on amide bonds in linear amides (3.5.1)
• CPC Class Codes: C12N 9/88 Lyases (4.)
• CPC Class Codes: C12N 9/93 Ligases (6)
• CPC Class Codes: C12P 13/001 Amines; Imines
• CPC Class Codes: C12P 13/10 Citrulline; Arginine; Ornithine
• CPC Class Codes: C12Y 201/03003 Ornithine carbamoyltransferase (2.1.3.3)
• CPC Class Codes: C12Y 203/01001 Amino-acid N-acetyltransferase (2.3.1.1)
• CPC Class Codes: C12Y 203/01008 Phosphate acetyltransferase (2.3.1.8)
• CPC Class Codes: C12Y 207/02001 Acetate kinase (2.7.2.1)
• CPC Class Codes: C12Y 305/01016 Acetylornithine deacetylase (3.5.1.16)
• CPC Class Codes: C12Y 401/01017 Ornithine decarboxylase (4.1.1.17)
• CPC Class Codes: C12Y 602/01001 Acetate-CoA ligase (6.2.1.1)

Abstract

Disclosed is a modified microorganism producing putrescine or ornithine, and a method for producing putrescine or ornithine using the same.

Claims

1. A modified microorganism of the genus Corynebacterium producing putrescine or ornithine, wherein activities of N-acetylglutamate synthase from E. coli and acetylornithine deacetylase from E. coli are introduced.

2. The modified microorganism according to claim 1, wherein the N-acetylglutamate synthase from E. coli consists of an amino acid sequence of SEQ ID NO:

1.

3. The modified microorganism according to claim 1, wherein the acetylornithine deacetylase from E. coli consists of an amino acid sequence of SEQ ID NO:

3.

4. The modified microorganism according to claim 1, wherein the microorganism of the genus Corynebacterium is Corynebacterium glutamicum.

5. The modified microorganism according to claim 1, wherein an activity of phosphotransacetylase and acetate kinase operon (pta-ackA operon) is further enhanced compared to its endogenous activity

6. The modified microorganism according to claim 5, wherein the phosphotransacetylase and acetate kinase operon consists of an amino acid sequence of SEQ ID NO:

5 or SEQ ID NO;

7.

7. The modified microorganism according to claim 1, wherein an activity of acetyl-CoA synthetase (acs) from E. coli is further introduced.

8. The modified microorganism according to claim 7, wherein the acetyl-CoA synthetase consists of an amino acid sequence of SEQ ID NO:

9.

9. The modified microorganism according to claim 1, wherein an activity of ornithine decarboxylase (ODC) is further introduced.

10. The modified microorganism according to claim 1, wherein an activity of i) ornithine carbamoyltransferase (ArgF), ii) glutamate exporter, or iii) ornithine carbamoyltransferase and glutamate exporter is further weakened compared to its endogenous activity.

11. The modified microorganism according to claim 1, wherein an activity of at least one selected from the group consisting of acetyl gamma glutamyl phosphate reductase (ArgC), acetylglutamate synthase or ornithine acetyltransferase (ArgJ), acetylglutamate kinase (ArgB), and acetyl ornithine aminotransferase (ArgD) is further enhanced compared to its endogenous activity.

12. The modified microorganism according to claim 1, wherein an activity of acetyltransferase is further weakened compared to its endogenous activity.

13. The modified microorganism according to claim 12, wherein the acetyltransferase consists of an amino acid sequence of SEQ ID NO:

30 or SEQ ID NO;

31.

14. The modified microorganism according to claim 1, wherein an activity of putrescine exporter is further enhanced compared to its endogenous activity.

15. The modified microorganism according to claim 14, wherein the putrescine exporter consists of an amino acid sequence of SEQ ID NO:

26 or SEQ ID NO;

28.

16. A method for producing putrescine or ornithine, comprising:

(i) culturing the modified microorganism of the genus Corynebacterium of claim 1 in a medium; and

(ii) recovering putrescine or ornithine from the cultured microorganism or the medium.

17. The method according to claim 16, wherein the microorganism of the genus Corynebacterium is Corynebacterium glutamicum.

18. The method according to claim 16, wherein the N-acetylglutamate synthase from E. coli consists of an amino acid sequence of SEQ ID NO:

1, and/or the acetylornithine deacetylase from E. coli consists of an amino acid sequence of SEQ ID NO;

3.

19. The method according to claim 16, wherein (a) an activity of phosphotransacetylase and acetate kinase operon (pta-ackA operon);

(b) an activity of at least one selected from the group consisting of acetyl gamma glutamyl phosphate reductase (ArgC), acetylglutamate synthase or ornithine acetyltransferase (ArgJ), acetylglutamate kinase (ArgB), and acetyl ornithine aminotransferase (ArgD); and

/or (c) an activity of putrescine exporter is further enhanced compared to its endogenous activity.

20. The method according to claim 16, wherein an activity of acetyl-CoA synthetase (acs) from E. coli, and/or an activity of ornithine decarboxylase (ODC) is further introduced.

21. The method according to claim 16, wherein (a) an activity of i) ornithine carbamoyltransferase (ArgF), ii) glutamate exporter, or iii) ornithine carbamoyltransferase and glutamate exporter and/or (b) an activity of acetyltransferase is further weakened compared to its endogenous activity.