Oxidase, polynucleotide encoding same, and use thereof

Patent metadata

Field	Value
Publication number	US-2017191097-A1
Application number	US-201515325190-A
Country	US
Kind code	A1
Filing date	Jul 2, 2015
Priority date	Jul 11, 2014
Publication date	Jul 6, 2017
Grant date	—

Abstract

Isolated oxidases, isolated polynucleotides encoding the oxidases, and methods of using the oxidases to produce α-oxocarboxylic acid compounds or L-α-amino acid compounds are described.

First claim

1 . A polynucleotide encoding any one of the following amino acid sequences (A1) to (A4):
(A1) an amino acid sequence represented by SEQ ID NO: 1, 3, or 5, (A2) an amino acid sequence i) having at least 45% sequence identity to an amino acid sequence represented by SEQ ID NO: 1, 3, or 5, and ii) having the ability to oxidize a 2-hydroxy-4-(methylthio)butyric acid derivative and convert the same into a corresponding 2-oxo-4-(methylthio)butyric acid derivative, (A3) an amino acid sequence i) encoded by a polynucleotide hybridized under a stringent condition to a polynucleotide composed of a sequence complementary to a base sequence represented by SEQ ID NO: 2, 4, or 6, and ii) of a protein having the ability to oxidize a 2-hydroxy-4-(methylthio)butyric acid derivative and convert the same into a corresponding 2-oxo-4-(methylthio)butyric acid derivative, or (A4) an amino acid sequence i) represented by SEQ ID NO: 1, 3, or 5 in which one or plural amino acids are deleted, substituted, or added, and ii) of a protein having the ability to oxidize a 2-hydroxy-4-(methylthio)butyric acid derivative and convert the same into a corresponding 2-oxo-4-(methylthio)butyric acid derivative.

2 . The polynucleotide according to claim 1 , which has a base sequence represented by SEQ ID NO: 2, 4, 6, 15, 16, or 17.

3 . A polynucleotide in which a promoter which can function in a host cell is connected with the polynucleotide according to claim 1 so that they can function.

4 . A recombinant vector comprising the polynucleotide according to claim 1 .

5 . The recombinant vector according to claim 4 , which further comprises a polynucleotide encoding an amino acid sequence of a protein having the ability to aminate an α-oxocarboxylic acid compound and convert the same into a corresponding L-α-amino acid compound, or a polynucleotide in which the polynucleotide is connected with a promoter which can function in a host cell so that they can function.

6 . The recombinant vector according to claim 5 , wherein the amino acid sequence of the protein having the ability to aminate an α-oxocarboxylic acid compound and convert the same into a corresponding L-α-amino acid compound is any one of the following amino acid sequences (B1) to (B3):
(B1) an amino acid sequence represented by SEQ ID NO: 7,
(B2) an amino acid sequence i) having at least 90% sequence identity to an amino acid sequence represented by SEQ ID NO: 7, and ii) of a protein having the ability to aminate a 2-oxo-4-(methylthio)butyric acid derivative and convert the same into a corresponding L-methionine derivative, or
(B3) an amino acid sequence i) represented by SEQ ID NO: 7 in which one or plural amino acids are deleted, substituted, or added, and ii) of a protein having the ability to aminate a 2-oxo-4-(methylthio)butyric acid derivative and convert the same into a corresponding L-methionine derivative.

7 . A transformant in which the recombinant vector according to claim 4 is introduced into a host cell.

8 . The transformant according to claim 7 , wherein the host cell is a microorganism or E. coli.

9 . A transformant in which the recombinant vector according to claim 5 is introduced into a host cell.

10 . The transformant according to claim 9 , wherein the host cell is a microorganism or E. coli.

11 . A transformant having the polynucleotide according to claim 1 .

12 . A transformant having the followings:
i) a polynucleotide having a base sequence encoding an amino acid sequence of a protein having the ability to aminate an α-oxocarboxylic acid compound and convert the same into a corresponding L-α-amino acid compound, or a polynucleotide in which the polynucleotide is connected with a promoter which can function in a host cell so that they can function; and ii) the polynucleotide according to claim 1 .

13 . A method for producing a recombinant vector, which comprises the step of integrating the polynucleotide according to claim 1 into a vector which can be replicated in a host cell.

14 . A method for producing a transformant, which comprises the step of introducing the polynucleotide according to claim 1 into a host cell.

15 . A protein having any one of the following amino acid sequences (A1) to (A4):
(A1) an amino acid sequence represented by SEQ ID NO: 1, 3, or 5, (A2) an amino acid sequence i) having at least 45% sequence identity to an amino acid sequence represented by SEQ ID NO: 1, 3, or 5, and ii) of a protein having the ability to oxidize a 2-hydroxy-4-(methylthio)butyric acid derivative and convert the same into a corresponding 2-oxo-4-(methylthio)butyric acid derivative, (A3) an amino acid sequence i) encoded by a polynucleotide hybridized under a stringent condition to a polynucleotide composed of a sequence complementary to a base sequence represented by SEQ ID NO: 2, 4, or 6, and ii) of a protein having the ability to oxidize a 2-hydroxy-4-(methylthio)butyric acid derivative and convert the same into a corresponding 2-oxo-4-(methylthio)butyric acid derivative, or (A4) an amino acid sequence i) represented by SEQ ID NO: 1, 3, or 5 in which one or plural amino acids are deleted, substituted, or added, and ii) of a protein having the ability to oxidize a 2-hydroxy-4-(methylthio)butyric acid derivative and convert the same into a corresponding 2-oxo-4-(methylthio)butyric acid derivative.

16 . A method for producing an α-oxocarboxylic acid compound, which comprises the step of reacting a protein having any one of the following amino acid sequences (A1) to (A4):
(A1) an amino acid sequence represented by SEQ ID NO: 1, 3, or 5,
(A2) an amino acid sequence i) having at least 45% sequence identity to an amino acid sequence represented by SEQ ID NO: 1, 3, or 5, and ii) of a protein having the ability to oxidize a 2-hydroxy-4-(methylthio)butyric acid derivative and convert the same into a corresponding 2-oxo-4-(methylthio)butyric acid derivative,
(A3) an amino acid sequence i) encoded by a polynucleotide hybridized under a stringent condition to a polynucleotide composed of a sequence complementary to a base sequence represented by SEQ ID NO: 2, 4, or 6, and ii) of a protein having the ability to oxidize a 2-hydroxy-4-(methylthio)butyric acid derivative and convert the same into a corresponding 2-oxo-4-(methylthio)butyric acid derivative, or
(A4) an amino acid sequence i) represented by SEQ ID NO: 1, 3, or 5 in which one or plural amino acids are deleted, substituted, or added, and ii) of a protein having the ability to oxidize a 2-hydroxy-4-(methylthio)butyric acid derivative and convert the same into a corresponding 2-oxo-4-(methylthio)butyric acid derivative;
with an α-hydroxycarboxylic acid compound.

17 . The production method according to claim 16 , wherein the α-hydroxycarboxylic acid compound is a sulfur-containing α-hydroxycarboxylic acid compound, and the corresponding α-oxocarboxylic acid compound is a sulfur-containing α-oxocarboxylic acid compound.

18 . The production method according to claim 17 , wherein the sulfur-containing α-hydroxycarboxylic acid compound is a compound represented by formula (1):

wherein R 1 represents a hydrogen atom or an optionally substituted a C1-8 alkyl group;
and the sulfur-containing α-oxocarboxylic acid compound is a compound represented by formula (2):

wherein R 1 is the same as defined above.

Assignees

• Sumitomo Chemical Co

Inventors

• Asako Hiroyuki

Classifications

• C12P11/00

  Preparation of sulfur-containing organic compounds · CPC title

• C12N5/10

  Cells modified by introduction of foreign genetic material · CPC title

• C12P7/40

  containing a carboxyl group {including Peroxycarboxylic acids} · CPC title

• C12Y104/01009

  Leucine dehydrogenase (1.4.1.9) · CPC title

• C12N15/09

  Recombinant DNA-technology · CPC title