Variant dihydrodipicolinate reductase polypeptide and method of producing L-threonine using the same

Patent metadata

Field	Value
Publication number	US-12497626-B2
Application number	US-202017623231-A
Country	US
Kind code	B2
Filing date	Aug 6, 2020
Priority date	Sep 26, 2019
Publication date	Dec 16, 2025
Grant date	Dec 16, 2025

Abstract

The present disclosure relates to a variant polypeptide having attenuated dihydrodipicolinate reductase activity and a method of producing L-threonine using the same.

First claim

The invention claimed is:

1 . A variant polypeptide having dihydrodipicolinate reductase activity, comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 1 with the exception of a substitution at position 13 with respect to the amino acid positions of SEQ ID NO: 1, wherein said substitution at position 13 is an asparagine, threonine, cysteine, tyrosine, serine, lysine, or glutamine, and wherein the dihydropicolinate reductase activity of the variant polypeptide is attenuated, as compared to said activity of the polypeptide having the amino acid sequence of SEQ ID NO: 1 or activity of the same variant polypeptide without the substitution at position 13.

2 . The variant polypeptide of claim 1 , wherein the amino acid sequence having at least 90% sequence identity to said amino acid sequence of SEQ ID NO: 1 is SEQ ID NO: 51.

3 . The variant polypeptide of claim 1 , wherein the variant polypeptide consists of the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 53.

4 . A polynucleotide encoding the variant polypeptide of claim 1 .

5 . The polynucleotide of claim 4 , wherein the polynucleotide consists of the nucleotide sequence of SEQ ID NO: 4 or SEQ ID NO: 54.

6 . A microorganism of the genus Corynebacterium comprising the variant polypeptide according to claim 1 .

7 . The microorganism of the genus Corynebacterium of claim 6 , wherein the amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 1 is SEQ ID NO: 51.

8 . The microorganism of the genus Corynebacterium of claim 6 , further comprising one or more of the variant polypeptides selected from the following variant polypeptides of (1) to (3): (1) a variant polypeptide having attenuated dihydrodipicolinate synthase activity; (2) a variant polypeptide having attenuated diaminopimelate decarboxylase activity; and (3) a variant polypeptide having attenuated diaminopimelate dehydrogenase activity.

9 . The microorganism of the genus Corynebacterium of claim 8 , wherein the variant polypeptide includes one or more of the variant polypeptides selected from the following variant polypeptides of (1) to (3): (1) a variant dihydrodipicolinate synthase polypeptide wherein tyrosine at position 119 of the amino acid sequence of SEQ ID NO: 65 is substituted with phenylalanine; (2) a variant diaminopimelate decarboxylase polypeptide wherein arginine at position 302 of the amino acid sequence of SEQ ID NO: 70 is substituted with alanine; and (3) a variant diaminopimelate dehydrogenase polypeptide wherein threonine at position 169 of the amino acid sequence of SEQ ID NO: 75 is substituted with leucine.

10 . The microorganism of the genus Corynebacterium of claim 6 , wherein the microorganism has increased L-threonine-producing ability, as compared with a non-modified strain.

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

12 . A method of producing L-threonine, the method comprising the step of culturing, in a medium, a microorganism of the genus Corynebacterium comprising the variant polypeptide according to claim 1 .

13 . The method of claim 12 , wherein the polypeptide sequence having at least 90% identity to SEQ ID NO: 1 is SEQ ID NO: 51.

14 . The method of claim 12 , further comprising the step of recovering L-threonine from the medium and microorganism cultured in the step of culturing the microorganism.

15 . A microorganism of the genus Corynebacterium comprising the polynucleotide according to claim 4 or a vector comprising the polynucleotide.

16 . A method of producing L-threonine, the method comprising the step of culturing, in a medium, a microorganism of the genus Corynebacterium comprising the polynucleotide according to claim 4 or a vector comprising the polynucleotide.

Assignees

• Cj Cheiljedang Corp

Inventors

• Baek Mina
• Kwon Su Yon
• Lee Imsang
• Son Seung-Ju
• Lee Kwang Woo

Classifications

• C12Y401/0102

  Diaminopimelate decarboxylase (4.1.1.20) · CPC title

• C12Y117/01

  with NAD+ or NADP+ as acceptor (1.17.1) · CPC title

• C12Y104/01016

  Diaminopimelate dehydrogenase (1.4.1.16) · CPC title

• C12Y103/01026

  Dihydrodipicolinate reductase (1.3.1.26) · CPC title

• C12P13/08

  Lysine; Diaminopimelic acid; Threonine; Valine · CPC title