O-succinyl homoserine transferase mutant and method for producing O-succinyl homoserine using same

Patent metadata

Field	Value
Publication number	US-10982245-B2
Application number	US-201816627572-A
Country	US
Kind code	B2
Filing date	Jun 29, 2018
Priority date	Jun 30, 2017
Publication date	Apr 20, 2021
Grant date	Apr 20, 2021

Abstract

The present disclosure relates to an O-succinyl homoserine transferase mutant, a polynucleotide encoding the same, a microorganism including the mutant, and a method of producing O-succinyl homoserine using the microorganism.

First claim

The invention claimed is:

1. An isolated variant polypeptide having O-succinyl homoserine transferase activity, which comprises a substitution of an amino acid at position 313 in the amino acid sequence of SEQ ID NO: 1 with arginine and a substitution of an amino acid at position 176 in the amino acid sequence of SEQ ID NO: 1 with an amino acid other than glutamine and wherein the variant has at least 90% sequence identity to SEQ ID NO:1.

2. The isolated polypeptide of claim 1 , wherein the amino acid at position 176 is substituted with asparagine, tryptophan, histidine, or glycine.

3. The isolated polypeptide of claim 1 , wherein the polypeptide comprises at least one amino acid sequence selected from the group consisting of amino acid sequences of SEQ ID NOs: 63, 75, 95, and 97.

4. An isolated polynucleotide encoding the polypeptide having the O-succinyl homoserine transferase activity of claim 1 .

5. The isolated polynucleotide of claim 4 , wherein the polynucleotide comprises at least one nucleic acid sequence selected from the group consisting of nucleic acid sequences of SEQ ID NOs: 64, 76, 96, and 98.

6. A microorganism of the genus Corynebacterium producing O-succinyl homoserine, wherein the microorganism comprises the polypeptide having the O-succinyl homoserine transferase activity of claim 1 or a mutant polypeptide thereof or wherein, in the microorganism, the polypeptide having the O-succinyl homoserine transferase activity of claim 1 or a mutant polypeptide thereof is overexpressed.

7. The microorganism of claim 6 , wherein the microorganism of the genus Corynebacterium has further enhanced aspartokinase activity compared to non-mutated microorganisms.

8. The microorganism of claim 6 , wherein the microorganism of the genus Corynebacterium is Corynebacterium glutamicum.

9. The microorganism of claim 6 , wherein activity of at least one enzyme selected from the group consisting of cystathionine synthase, O-acetyl homoserine (thiol)-lyase, and homoserine kinase is inactivated in the microorganism of the genus Corynebacterium.

10. The microorganism of claim 6 , wherein the microorganism of the genus Corynebacterium has further enhanced aspartokinase activity compared to non-mutated microorganisms and activity of at least one enzyme selected from the group consisting of cystathionine synthase, O-acetyl homoserine (thiol)-lyase, and homoserine kinase is inactivated.

11. A method of producing O-succinyl homoserine, the method comprising:
culturing a microorganism of the genus Corynebacterium producing O-succinyl homoserine according to claim 6 in a culture medium; and
separating or recovering O-succinyl homoserine from the microorganism cultured in the culturing step or the culture medium.

12. A method of producing L-methionine, the method comprising:
(a) culturing a microorganism of the genus Corynebacterium producing O-succinyl homoserine according to claim 6 in a culture medium; and
(b) reacting the O-succinyl homoserine with a sulfide.

13. The method of claim 12 , further comprising separating or recovering O-succinyl homoserine from the cultured microorganism or the culture medium in step (a).

14. The method of claim 12 , further comprising separating or recovering L-methionine produced by the reaction between O-succinyl homoserine and the sulfide in step (b).

Assignees

• Cj Cheiljedang Corp

Inventors

• Kim Kyungrim
• Shim Jihyun
• Kim Hyun Ah
• Shin Yong Uk
• Lee Peter

Classifications

• C12Y203/01031

  Homoserine O-acetyltransferase (2.3.1.31) · CPC title

• C12R2001/15

  Corynebacterium · CPC title

• C12N15/77 Primary

  for Corynebacterium; for Brevibacterium · CPC title

• C12N15/1034

  Isolating an individual clone by screening libraries · CPC title

• C12N9/1029

  transferring groups other than amino-acyl groups (2.3.1) · CPC title