Method for producing methacrylyl-coa

Patent metadata

Field	Value
Publication number	US-2016145665-A1
Application number	US-201414905155-A
Country	US
Kind code	A1
Filing date	Jul 25, 2014
Priority date	Aug 1, 2013
Publication date	May 26, 2016
Grant date	—

Abstract

The invention provides a method for producing methacrylyl-CoA that converts 3-hydroxyisobutyryl-CoA into methacrylyl-CoA using an enzyme having dehydratase activity as a method for producing methacrylyl-CoA using an enzyme catalyst. In this production method, conversion rate of 3-hydroxyisobutyryl-CoA into methacrylyl-CoA by the enzyme having dehydratase activity is 50% or higher. In this production method, furthermore, the enzyme having dehydratase activity derives from a microorganism belonging to the genus Pseudomonas or Rhodococcus.

First claim

1 . A method for producing methacrylyl-CoA, the method comprising:
converting 3-hydroxyisobutyryl-CoA to methacrylyl-CoA in the presence of a dehydratase.

2 . The method according to claim 1 , wherein a hydratase with a conversion rate of 50% or higher is used for converting 3-hydroxyisobutyryl-CoA to methacrylyl-CoA.

3 . The method according to claim 1 , wherein 3-hydroxyisobutyryl-CoA is converted to methacrylyl-CoA at a pH of 4˜10.

4 . The method according to claim 1 , wherein 3-hydroxyisobutyryl-CoA is converted to methacrylyl-CoA at a temperature of 5˜80° C.

5 . The method according to claim 1 , wherein 3-hydroxyisobutyryl-CoA is converted to methacrylyl-CoA for a duration of from 1 minute to 1 week.

6 . The method according to claim 1 , wherein 3-hydroxyisobutyryl-CoA is prepared in an aqueous medium comprising 1 mM or greater of an osmolyte.

7 . The method according to claim 1 , wherein 3-hydroxyisobutyryl-CoA is converted to methacrylyl-CoA in the presence of a transformant for expressing a gene for encoding a dehydratase.

8 . The method according to claim 7 , wherein the gene encoding a dehydratase is derived from a microorganism.

9 . The method according to claim 8 , wherein the microorganism belongs to the genus Pseudomonas or the genus Rhodococcus.

10 . The method according to claim 7 , wherein the dehydratase is selected from a group consisting of (a)˜(f) below:
(a) a protein having an amino acid sequence shown in SEQ ID NO: 1 or 3;
(b) a protein having an amino acid sequence in which one or more amino acids are deleted from, substituted with, added to and/or inserted into the amino acid sequence shown in SEQ ID NO: 1 or 3, and possessing dehydratase activity;
(c) a protein being 90% or greater identical to a protein having the amino acid sequence shown in SEQ ID NO: 1 or 3, and possessing dehydratase activity;
(d) a protein encoded by DNA having a base sequence shown in SEQ ID NO: 2 or 4;
(e) a protein encoded by DNA that hybridizes with a DNA strand having the base sequence shown in SEQ ID NO: 2 or 4 under stringent conditions, and possessing dehydratase activity; and
(f) a protein encoded by DNA that is 90% or greater identical to a DNA strand having the base sequence shown in SEQ ID NO: 2 or 4, and possessing dehydratase activity.

11 . A method for producing methacrylyl-CoA the method comprising:
converting 3-hydroxyisobutyryl-CoA to methacrylyl-CoA in the presence of a transformant for expressing a gene encoding a dehydratase derived from a microorganism at a conversion rate of 50% or higher through reactions of a 3-hydroxyisobutyryl-CoA solution, which is prepared in an aqueous medium comprising 1 mM or greater of an osmolyte, at a pH of 4˜10 and a temperature of 5˜80° C. for a duration of from 1 minute to 1 week.

Assignees

• Mitsubishi Rayon Co

Inventors

• Sato Eiji
• Yu Fujio
• Nakajima Eiji
• Yamazaki Michiko
• Mizunashi Wataru

Classifications

• C12N9/88

  Lyases (4.) · CPC title

• C12P19/32 Primary

  having a condensed ring system containing a six-membered ring having two N-atoms in the same ring, e.g. purine nucleotides, nicotineamide-adenine dinucleotide · CPC title

• C12P7/52

  Propionic acid; Butyric acids · CPC title