Hydrolase and method for producing (1S,2S)-1-alkoxycarbonyl-2-vinylcyclopropane carboxylic acid using same
AssigneeUbe Corporation
InventorsToyokazu Yoshida, Koichi Ishida, Ryoma Miyake, Takanobu Iura, Hiroshi Kawabata
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Abstract
The present invention provides a novel hydrolase that can industrially produce optically highly pure (1S,2S)-1-alkoxycarbonyl-2-vinylcyclopropane carboxylic acid with high efficiency at low costs, and a production method using the hydrolase.
Claims7
A method for producing (1S,2S)-1-alkoxycarbonyl-2-vinylcyclopropane carboxylic acid, comprising bringing a hydrolase, a microorganism or cell having an ability to produce the hydrolase, a and/or a culture solution containing the hydrolase into contact with dialkyl 2-vinylcyclopropane-1,1-dicarboxylic acid represented by formula (I) to produce (1S,2S)-1-alkoxycarbonyl-2-vinylcyclopropane carboxylic acid represented by formula (II),
- wherein the hydrolase comprises a polypeptide of the following (a), (b), or (c):
- (a) a polypeptide consisting of an amino acid sequence resulting from the deletion, insertion, substitution, and/or addition of 1 to 50 amino acids in the amino acid sequence shown in SEQ ID NO:
- 4 or 6, and an activity to catalyze the reaction shown in the formula (1):
- wherein R is an alkyl group having 1 to 6 carbon atoms, and a ratio of Syn isomer to Anti isomer in formula (II) is equal to or less than 0.2%;
- (b) a polypeptide consisting of an amino acid sequence with not less than 95% sequence identity with the amino acid sequence shown in SEQ ID NO:
- 4 or 6, and an activity to catalyze the reaction shown in formula (1);
- (c) a polypeptide consisting of an amino acid sequence with not less than 90% sequence identity with the amino acid sequence shown in SEQ ID NO:
- 4 or 6, wherein the amino acid sequence comprises:
- (i) 5 consecutive residues DATRG corresponding to residues 75-79 of SEQ ID NO:
- 4 and residues 80-84 of SEQ ID NO:
- 6;
- ii 4 consecutive residues PYGF corresponding to residues 132-135 of SEQ ID NO:
- 4 and residues 136-139 of SEQ ID NO:
- 6;
- iii 4 consecutive residues NWPG corresponding to residues 222-225 of SEQ ID NO:
- 4 and residues 226-229 of SEQ ID NO:
- 6;
- and/or
- (iv) 5 consecutive residues PGWPG corresponding to residues 285-289 of SEQ ID NO:
- 4 and residues 289-293 of SEQ ID NO:
- 6.
A method for producing (1S,2S)-1-alkoxycarbonyl-2-vinylcyclopropane carboxylic acid, comprising bringing a treated product of a microorganism or cell having the ability to produce a hydrolase into contact with dialkyl 2-vinylcyclopropane-1,1-dicarboxylic acid represented by formula (I) to produce (1S,2S)-1-alkoxycarbonyl-2-vinylcyclopropane carboxylic acid represented by formula (II),
- wherein the hydrolase comprises a polypeptide of the following (a), (b), or (c):
- (a) a polypeptide consisting of an amino acid sequence resulting from the deletion, insertion, substitution, and/or addition of 1 to 50 amino acids in the amino acid sequence shown in SEQ ID NO:
- 4 or 6, and an activity to catalyze the reaction shown in formula (1):
- wherein R is an alkyl group having 1 to 6 carbon atoms, and a ratio of Syn isomer to Anti isomer in formula (II) is equal to or less than 0.2%;
- (b) a polypeptide consisting of an amino acid sequence with not less than 95% sequence identity with the amino acid sequence shown in SEQ ID NO:
- 4 or 6, and an activity to catalyze the reaction shown in formula (1);
- (c) a polypeptide consisting of an amino acid sequence with not less than 90% sequence identity with the amino acid sequence shown in SEQ ID NO:
- 4 or 6, wherein the amino acid sequence comprises:
- (i) 5 consecutive residues DATRG corresponding to residues 75-79 of SEQ ID NO:
- 4 and residues 80-84 of SEQ ID NO:
- 6;
- (ii) 4 consecutive residues PYGF corresponding to residues 132-135 of SEQ ID NO:
- 4 and residues 136-139 of SEQ ID NO:
- 6;
- (iii) 4 consecutive residues NWPG corresponding to residues 222-225 of SEQ ID NO:
- 4 and residues 226-229 of SEQ ID NO:
- 6;
- and/or
- (iv) 5 consecutive residues PGWPG corresponding to residues 285-289 of SEQ ID NO:
- 4 and residues 289-293 of SEQ ID NO:
- 6.
Dependencies are inferred from the claim text.