Compositions and methods of producing methionine

Patent metadata

Field	Value
Publication number	US-9493801-B2
Application number	US-201514820453-A
Country	US
Kind code	B2
Filing date	Aug 6, 2015
Priority date	Apr 11, 2007
Publication date	Nov 15, 2016
Grant date	Nov 15, 2016

Abstract

Described herein are microorganisms that produce methionine and related products from endogenous genes in a transsulfuration pathway, as well as from exogenous genes providing a direct sulfhydrylation pathway. Novel genes that are useful for methionine and SAMe production are disclosed.

First claim

What is claimed is:

1. An isolated polynucleotide encoding an isolated polypeptide with homoserine O-suceinyltransferase activity, wherein the isolated polypeptide shows a reduced sensitivity to feedback inhibition by L-methionine as compared with a wild-type homoserine O-succinyltransferase polypeptide having the amino acid sequence of SEQ ID NO: 35, and comprises a mutation at one or more amino acid positions corresponding to amino acids 24, 29, 79, 114, 140, 163, 222, 275, 290, 291, 295, 297, 304 and 305 of SEQ ID NO: 35.

2. The isolated polynucleotide of claim 1 , wherein the isolated polypeptide comprises a mutation at one or more amino acid positions corresponding to amino acids 163, 222 and 295 of SEQ ID NO: 35.

3. The isolated polynucleotide of claim 1 , wherein the isolated polypeptide comprises a mutation at one or more amino acid positions corresponding to amino acids 24, 275, 297 and 305 of SEQ ID NO: 35.

4. The isolated polynucleotide of claim 1 , wherein the isolated polypeptide comprises a mutation at an amino acid position corresponding to an amino acid 290 of SEQ ID NO: 35.

5. The isolated polynucleotide of claim 1 , wherein the isolated polypeptide comprises a mutation at one or more amino acid positions corresponding to amino acids 29, 114 and 140 of SEQ ID NO: 35.

6. The isolated polynucleotide of claim 1 , wherein the isolated polypeptide comprises a mutation at one or more amino acid positions corresponding to amino acids 79, 140, 291 and 304 of SEQ ID NO: 35.

7. The isolated polynucleotide of claim 1 , wherein in the amino acid sequence of SEQ ID NO: 35, threonine at position 24 is replaced with serine; serine at position 29 is replaced with proline; asparagine at position 79 is replaced with serine; glutamic acid at 114 is replaced with glycine; phenylalanine at position 140 is replaced with serine or isoleucine; lysine at position 163 is replaced with glutamine; phenylalanine at position 222 is replaced with leucine; alanine at position 275 is replaced with glutamic acid; asparagine at position 290 is replaced with histidine; tyrosine at position 291 is replaced with asparagine; glutamine at position 295 is replaced With arginine; threonine at position 297 is replaced with alanine; methionine at position 304 is replaced with leucine; and asparagine at position 305 is replaced with tyrosine.

8. The isolated polynucleotide of claim 1 , wherein in the amino acid sequence of SEQ ID NO: 35, lysine at position 163 is replaced with glutamine; phenylalanine at position 222 is replaced with leucine; and glutamine at position 295 is replaced with arginine.

9. The isolated polynucleotide of claim 1 , wherein in the amino acid sequence of SEQ ID NO: 35, threonine at position 24 is replaced with serine; alanine at position 275 is replaced with glutamic acid; threonine at position 297 is replaced with alanine; and asparagine at position 305 is replaced with tyrosine.

10. The isolated polynucleotide of claim 1 , wherein in the amino acid sequence of SEQ ID NO: 35, asparagine at position 290 is replaced with histidine.

11. The isolated polynucleotide of claim 1 , wherein in the amino acid sequence of SEQ ID NO: 35, serine at position 29 is replaced with proline; glutamic acid at position 114 is replaced with glycine; and phenylalanine at position 140 is replaced with serine or isoleucine.

12. The isolated polynucleotide of claim 1 , wherein in the amino acid sequence of SEQ ID NO: 35, asparagine at position 79 is replaced with serine; phenylalanine at position 140 is replaced with serine or isoleucine; tyrosine at position 291 is replaced with asparagine; and methionine at position 304 is replaced with leucine.

13. The isolated polynucleotide of claim 1 , wherein the isolated polypeptide comprises the amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 4, 6, 8 and 10.

14. The isolated polynucleotide of claim 1 , wherein the isolated polynucleotide comprises the polynucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7 and 9.

15. A microorganism strain modified to comprise the isolated polynucleotide according to claim 1 .

16. A method of producing L-methinine comprising: culturing the microorganism according to claim 15 under conditions that permit production of L-methionine; and isolating thus produced L-methionine.

Assignees

• Cj Cheiljedang Corp

Inventors

• Brazeau Brian
• Chang Jin-Sook
• Cho Kwang Myung
• Cho Young Wook
• Desouza Mervyn
• Jessen Holly J
• Kim So-Young
• Niu Wei
• Sanchez-Riera Fernando A
• Shin Yong-Uk
• Um Hyewon

Classifications

• C12N15/63

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title

• C12P13/12 Primary

  Methionine; Cysteine; Cystine · CPC title

• C12Y203/01046

  Homoserine O-succinyltransferase (2.3.1.46) · CPC title

• C12N9/1029

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

• Y02P20/52

  using catalysts, e.g. selective catalysts · CPC title