Amidase, gene for the same, vector, transformant, and method for production of optically active carboxylic acid amide and optically active carboxylic acid by using any one of those items

Patent metadata

Field	Value
Publication number	US-9783796-B2
Application number	US-53229008-A
Country	US
Kind code	B2
Filing date	Mar 14, 2008
Priority date	Mar 22, 2007
Publication date	Oct 10, 2017
Grant date	Oct 10, 2017

Abstract

The present invention has its object to provide a novel polypeptide having amidase activity to selectively hydrolyze S-enantiomer in racemic nipecotamide, a DNA encoding the polypeptide, a vector containing the DNA, a transformant transformed with the vector, and a method for producing an optically active carboxylic acid amide and an optically active carboxylic acid in which a racemic carboxylic acid amide is hydrolyzed with the polypeptide or the transformant.

First claim

The invention claimed is:

1. An isolated DNA which encodes a polypeptide which is any one of the following polypeptides (b), and (c):
(b) a polypeptide comprising an amino acid sequence derived from the amino acid sequence set forth in SEQ ID NO:1 by substitution, insertion, deletion, and/or addition of 90 or less amino acids, and having activity to selectively hydrolyze S-enantiomer in racemic nipecotamide and wherein the polypeptide is not SEQ ID NO: 1; and
(c) a polypeptide having sequence identity of 80% or higher to the amino acid sequence set forth in SEQ ID NO:1, and having activity to selectively hydrolyze S-enantiomer in racemic nipecotamide and wherein the polypeptide is not SEQ ID NO: 1.

2. An isolated DNA which is any one of the following DNAs (a), (b), and (c):
(b) a DNA hybridizable under a stringent condition with a DNA comprising a nucleotide sequence complementary to the nucleotide sequence set forth in SEQ ID NO:2, and encoding a polypeptide having activity to selectively hydrolyze S-enantiomer in racemic nipecotamide, and wherein the DNA does not comprise SEQ ID NO: 2,
wherein the stringent condition is a condition where hybridization is carried out in the presence of 0.7 to 1.0 M NaCl, at 65° C., followed by washing the filter with 0.2-fold concentration of SSC solution at 65° C.; and
(c) a DNA having sequence identity of 80% or higher to the nucleotide sequence set forth in SEQ ID NO:2, and encoding a polypeptide having activity to selectively hydrolyze S-enantiomer in racemic nipecotamide, and wherein the DNA does not comprise SEQ ID NO: 2.

3. The DNA according to claim 2 , which has sequence identity of 85% or higher to the nucleotide sequence set forth in SEQ ID NO:2, and encodes the polypeptide having activity to selectively hydrolyze S-enantiomer in racemic nipecotamide.

4. A vector comprising an isolated DNA which encodes a polypeptide which is any one of the following polypeptides (a), (b), and (c):
(a) a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:1;
(b) a polypeptide comprising an amino acid sequence derived from the amino acid sequence set forth in SEQ ID NO:1 by substitution, insertion, deletion, and/or addition of 90 or less amino acids, and having activity to selectively hydrolyze S-enantiomer in racemic nipecotamide; and
(c) a polypeptide having sequence identity of 80% or higher to the amino acid sequence set forth in SEQ ID NO:1, and having activity to selectively hydrolyze S-enantiomer in racemic nipecotamide.

5. A transformant which is producible by transformation of a host microorganism with the vector according to claim 4 , wherein said host microorganism is not Cupriavidus sp.

6. The transformant according to claim 5 , wherein the host microorganism is Escherichia coli.

7. The isolated DNA according to claim 1 which encodes the polypeptide which has sequence identity of 85% or higher to the amino acid sequence set forth in SEQ ID NO:1, and has activity to selectively hydrolyze S-enantiomer in racemic nipecotamide.

8. The DNA according to claim 1 , which is isolated from a microorganism belonging to the genus Cupriavidus.

9. The DNA according to claim 8 , wherein the microorganism belonging to the genus Cupriavidus is Cupriavidus sp. KNK-J915 strain (FERM BP-10739).

10. A vector comprising an isolated DNA which is any one of the following DNAs (a), (b), and (c):
(a) a DNA comprising the nucleotide sequence set forth in SEQ ID NO:2;
(b) a DNA hybridizable under a stringent condition with a DNA comprising a nucleotide sequence complementary to the nucleotide sequence set forth in SEQ ID NO:2, and encoding a polypeptide having activity to selectively hydrolyze S-enantiomer in racemic nipecotamide,
wherein the stringent condition is a condition where hybridization is carried out in the presence of 0.7 to 1.0 M NaCl, at 65° C., followed by washing the filter with 0.2-fold concentration of SSC solution at 65° C.; and
(c) a DNA having sequence identity of 80% or higher to the nucleotide sequence set forth in SEQ ID No:2, and encoding a polypeptide having activity to selectively hydrolyze S-enantiomer in racemic nipecotamide.

Assignees

• Nojiri Masutoshi
• Moriyama Daisuke
• Nishiyama Tozo
• Taoka Naoaki
• Kaneka Corp

Inventors

• Nojiri Masutoshi
• Moriyama Daisuke
• Nishiyama Tozo
• Taoka Naoaki

Classifications

• C12P41/006

  by reactions involving C-N bonds, e.g. nitriles, amides, hydantoins, carbamates, lactames, transamination reactions, or keto group formation from racemic mixtures · CPC title

• C12P13/02

  Amides, e.g. chloramphenicol {or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes (peptides C12P21/00 or C07K)} · CPC title

• C12N9/80 Primary

  acting on amide bonds in linear amides {(3.5.1)} · CPC title