Method for producing optically active amine compounds by deracemization

Patent metadata

Field	Value
Publication number	US-9777301-B2
Application number	US-201314352269-A
Country	US
Kind code	B2
Filing date	Dec 13, 2013
Priority date	Dec 13, 2012
Publication date	Oct 3, 2017
Grant date	Oct 3, 2017

Abstract

Disclosed are methods for producing optically active amino acids and amines. According to the methods, α-keto acids are generated as reaction intermediates, and as a result, ω-transaminase-catalyzed kinetic resolution of racemic amino acids or amines as racemic amine compounds enables the production of optically active amine compounds without the need to use expensive α-keto acids as starting materials. Therefore, the optically active amine compounds are produced at greatly reduced costs. In addition, the optically active amine compounds have high enantiomeric excess.

First claim

We claim:

1. A one-pot method for producing an optically pure amino acid by deracemizing a racemic amino acid with two transaminases of opposite enantioselectivity and a co-substrate dedicated for each transaminase, comprising:
1) providing a racemic amino acid, an amino acceptor for an amino acid transaminase, and an amino donor for a co-transaminase, to the amino acid transaminase and the ω-transaminase whose enantioselectivities are opposite to each other;
2) contacting the racemic amino acid, the amino acceptor, and the amino acid transaminase provided in step 1) to convert one amino acid enantiomer of the racemic amino acid to an α-keto acid; and
3) contacting the α-keto acid produced in step 2), the amino donor provided in step 1), and the ω-transaminase provided in step 1) to convert the α-keto acid to the amino acid enantiomer whose chirality is opposite to the amino acid enantiomer reacted in step 2),
wherein an (R)-amino acid transaminase and a (S)-selective ω-transaminase are used as the amino acid transaminase and the ω-transaminase, respectively, to deracemize the racemic amino acid into optically pure (S)-amino acid, or
wherein a (S)-amino acid transaminase and an (R)-selective ω-transaminase are used as the amino acid transaminase and the ω-transaminase, respectively, to deracemize the racemic amino acid into optically pure (R)-amino acid.

2. The one-pot method according to claim 1 , wherein the (R)-amino acid transaminase is provided in step 1), the (R)-amino acid of the racemic amino acid and the amino acceptor are contacted with the R-amino acid transaminase to produce an α-keto acid in step 2), and the α-keto acid is contacted with the (S)-selective ω-transaminase to produce a (S)-amino acid of the α-keto acid in step 3).

3. The one-pot method according to claim 1 , wherein the (S)-amino acid transaminase is provided in step 1), the (S)-amino acid of the racemic amino acid and the amino acceptor are contacted with the (S)-amino acid transaminase to produce an α-keto acid in step 2), and the a-keto acid is contacted with the (R)-selective ω-transaminase to produce an (R)-amino acid of the a-keto acid in step 3).

4. The one-pot method according to claim 1 , wherein the (S)-selective ω-transaminase is an enzyme derived from Ochrobactrum anthropi and encoded by the nucleic acid sequence set forth in SEQ ID NO: 1 or an enzyme derived from Paracoccus denitrificans and encoded by the nucleic acid sequence set forth in SEQ ID NO: 2.

5. The one-pot method according to claim 1 , wherein the (R)-selective co-transaminase is an enzyme derived from Aspergillus terreus and encoded by the nucleic acid sequence set forth in SEQ ID NO: 3 or an enzyme derived from Arthrobacter sp. and encoded by the nucleic acid sequence set forth in SEQ ID NO: 4.

6. The one-pot method according to claim 1 , wherein the racemic amino acid is selected from the group consisting of racemic alanine, racemic serine, racemic homoserine, racemic norvaline, racemic norleucine, racemic leucine, and mixtures thereof.

7. The one-pot method according to claim 1 , wherein the amino acceptor is selected from the group consisting of a-ketoglutarate, phenylglyoxylate, phenylpyruvate, and mixtures thereof.

8. The one-pot method according to claim 1 , wherein the amino donor is selected from the group consisting of isopropylamine, methylbenzylamine, benzylamine, and mixtures thereof.

9. The one-pot method according to claim 1 , wherein the (R)-selective amino acid transaminase is an enzyme derived from Bacillus sphaericus and encoded by the nucleic acid sequence set forth in SEQ ID NO: 5.

10. The one-pot method according to claim 1 , wherein the (R)-selective amino acid transaminase is an enzyme derived from Escherichia coli and encoded by the nucleic acid sequence set forth in SEQ ID NO: 6.

Assignees

• Industry-Academic Cooperation Foundation Yonsei Univ

Inventors

• Shin Jong-Shik
• Park Eul-Soo
• Malik M Shaheer

Classifications

• C12P13/06

  Alanine; Leucine; Isoleucine; Serine; Homoserine · CPC title

• C12P13/001 Primary

  Amines; Imines · CPC title

• C12P41/007

  by reactions involving acyl derivatives of racemic amines · CPC title