Method for producing L-α-amino acid compound

Patent metadata

Field	Value
Publication number	US-10364448-B2
Application number	US-201515325190-A
Country	US
Kind code	B2
Filing date	Jul 2, 2015
Priority date	Jul 11, 2014
Publication date	Jul 30, 2019
Grant date	Jul 30, 2019

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

The invention claimed is:

1. A method for producing a sulfur containing L-α-amino acid compound of formula (3):

wherein the method comprises:
(I) a step of reacting a protein with a sulfur-containing α-hydroxycarboxylic acid compound of formula (1) to obtain the corresponding sulfur-containing α-oxocarboxylic acid compound of formula (2):

wherein the protein is selected from the group consisting of:
(A1) a protein having the amino acid sequence of SEQ ID NO: 5,
(A2) a protein having an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 5, wherein the protein has the ability to oxidize and convert the sulfur-containing α-hydroxycarboxylic acid compound of formula (1) into the corresponding sulfur-containing α-oxocarboxylic acid compound of formula (2), and
(A3) a protein having the amino acid sequence of SEQ ID NO: 5 in which 1 to 10 amino acids are deleted, substituted, or added, wherein the protein has the ability to oxidize and convert the sulfur-containing α-hydroxycarboxylic acid compound of formula (1) into the corresponding sulfur-containing α-oxocarboxylic acid compound of formula (2); and
(II) a step of reacting the sulfur-containing α-oxocarboxylic acid compound of formula (2) obtained in the step (I) with a protein having the ability to aminate and convert the sulfur-containing α-oxocarboxylic acid compound of formula (2) into the corresponding sulfur-containing L-α-amino acid compound of formula (3) to thereby obtain the sulfur-containing L-α-amino acid compound of formula (3), wherein the protein is selected from the group consisting of:
(B1) a protein having the amino acid sequence of SEQ ID NO: 7,
(B2) a protein having an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 7, wherein the protein has the ability to aminate and convert the sulfur-containing α-oxocarboxylic acid compound of formula (2) into the corresponding sulfur-containing L-α-amino acid compound of formula (3), and
(B3) a protein having the amino acid sequence of SEQ ID NO: 7 in which 1 to 36 amino acids are deleted, substituted, or added, wherein the protein has the ability to aminate and convert the sulfur-containing α-oxocarboxylic acid compound of formula (2) into the corresponding sulfur-containing L-α-amino acid compound of formula (3),
wherein R 1 in formula (1), formula (2), and formula (3) is a C 1 -C 8 alkyl group.

2. The production method according to claim 1 , wherein the protein in step (II) having the ability to aminate and convert the sulfur-containing α-oxocarboxylic acid compound of formula (2) into the corresponding sulfur-containing L-α-amino acid compound of formula (3) is a leucine dehydrogenase.

3. The production method according to claim 2 , wherein the leucine dehydrogenase is a leucine dehydrogenase derived from Bacillus sphaericus.

4. The production method according to claim 1 , wherein the protein having any one of the amino acid sequences (A1), (A2), or (A3) is provided as a transformant, a freeze-dried transformant, an organic solvent-treated transformant, a dried transformant, a triturated transformant, an autolysate of a transformant, a sonicate of a transformant, a transformant extract, or an alkali-treated product of a transformant, wherein the transformant is a host cell comprising a polynucleotide encoding the protein.

5. The production method according to claim 4 , wherein the polynucleotide is comprised in a recombinant vector.

6. The production method according to claim 1 , wherein the protein having any one of the amino acid sequences (B1), (B2), or (B3) is provided as a transformant, a freeze-dried transformant, an organic solvent-treated transformant, a dried transformant, a triturated transformant, an autolys ate of a transformant, a sonic ate of a transformant, a transformant extract, or an alkali-treated product of a transformant, wherein the transformant is a host cell comprising a polynucleotide encoding the protein.

7. The production method according to claim 6 , wherein the polynucleotide is comprised in a recombinant vector.

8. The production method according to claim 1 , wherein step (I) is performed in the presence of a protein having the ability to convert hydrogen peroxide into molecular oxygen.

9. The production method according to claim 8 , wherein the protein having the ability to convert hydrogen peroxide into molecular oxygen is a catalase.

10. The production method according to claim 8 , wherein the protein having the ability to convert hydrogen peroxide into molecular oxygen is provided as a transformant a freeze-dried transformant, an organic solvent-treated transformant, a dried transformant, a triturated transformant, an autolysate of a transformant, a sonicate of a transformant, a transformant extract, or an alkali-treated product of a transformant, wherein the transformant is a host cell comprising a polynucleotide encoding the protein.

11. The production method according to claim 1 , wherein step (II) is performed in the presence of a protein having the ability to convert an oxidized β-nicotinamide adenine dinucleotide or an oxidized β-nicotinamide adenine dinucleotide phosphate into its reduced form.

12. The production method according to claim 11 , wherein the protein having the ability to convert an oxidized β-nicotinamide adenine dinucleotide or an oxidized β-nicotinamide adenine dinucleotide phosphate into its reduced form is a formate dehydrogenase.

13. The production method according to claim 11 , wherein the protein having the ability to convert an oxidized β-nicotinamide adenine dinucleotide or an oxidized β-nicotinamide adenine dinucleotide phosphate into its reduced form is provided as a transformant, a freeze-dried transformant, an organic solvent-treated transformant, a dried transformant, a triturated transformant, an autolysate of a transformant, a sonicate of a transformant, a transformant extract, or an alkali-treated product of a transformant, wherein the transformant is a host cell comprising a polynucleotide encoding the protein.

14. The production method according to claim 1 , wherein step (I) and step (II) are performed in a single reaction system.

15. The production method according to claim 1 , wherein the sulfur containing L-α-amino acid compound of formula (3) is L-methionine.

Assignees

• Sumitomo Chemical Co

Inventors

• Asako Hiroyuki

Classifications

• C12P9/00

  Preparation of organic compounds containing a metal or atom other than H, N, C, O, S or halogen {(phosphoglycerides, C12P7/6481)} · CPC title

• C12N15/09

  Recombinant DNA-technology · CPC title

• C12P11/00

  Preparation of sulfur-containing organic compounds · CPC title

• C12Y104/01009

  Leucine dehydrogenase (1.4.1.9) · CPC title

• C12P7/40

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