Method for producing methacrylyl-CoA
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.
1. A method for producing methacrylyl-CoA and converting methylacryl-CoA to methacrylic acid or methacrylate ester, the method comprising:
converting 3-hydroxyisobutyryl-CoA to methacrylyl-CoA in an aqueous medium in the presence of a dehydratase at a conversion rate of 50% or higher; and
converting methacrylyl-CoA to methacrylic acid or methacrylate ester at a pH of 4 to 10,
wherein the dehydratase is selected from a group consisting of:
(a) a protein having the amino acid sequence shown in SEQ ID NO: 1 or 3;
(b) a protein that is at least 80% identical to a protein having the amino acid sequence shown in SEQ ID NO: 1 or 3, and possessing dehydratase activity;
(c) a protein encoded by the nucleotide sequence of SEQ ID NO: 2 or 4;
(d) a protein encoded by DNA that is at least 80% identical to SEQ ID NO: 2 or 4, and possessing dehydratase activity.
2. The method according to claim 1 , wherein 3-hydroxyisobutyryl-CoA is converted to methacrylyl-CoA at a temperature of 5˜80° C.
3. 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.
4. The method according to claim 1 , wherein the aqueous medium comprises 1 mM or greater of an osmolyte.
5. 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.
6. The method according to claim 5 , wherein the gene encoding a dehydratase is derived from a microorganism.
7. The method according to claim 6 , wherein the microorganism belongs to the genus Pseudomonas or the genus Rhodococcus.
8. The method according to claim 1 , wherein the dehydratase has the amino acid sequence shown in SEQ ID NO: 1 or 3.
9. 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,
wherein the dehydratase is selected from a group consisting of:
(a) a protein having the amino acid sequence shown in SEQ ID NO: 1 or 3;
(b) a protein that is at least 80% identical to a protein having the amino acid sequence shown in SEQ ID NO: 1 or 3, and possessing dehydratase activity;
(c) a protein encoded by the nucleotide sequence of SEQ ID NO: 2 or 4;
(d) a protein encoded by DNA that is at least 80% identical to SEQ ID NO: 2 or 4, and possessing dehydratase activity.
10. The method according to claim 9 , wherein the dehydratase has the amino acid sequence shown in SEQ ID NO: 1 or 3.