US 12,258,593 B2
Hydrolase and method for producing (1S,2S)-1-alkoxycarbonyl-2-vinylcyclopropane carboxylic acid using same
Toyokazu Yoshida, Gifu (JP); Koichi Ishida, Gifu (JP); Ryoma Miyake, Tokyo (JP); Takanobu Iura, Tokyo (JP); and Hiroshi Kawabata, Tokyo (JP)
Assigned to UBE CORPORATION, Yamaguchi (JP)
Filed by UBE CORPORATION, Yamaguchi (JP)
Filed on Dec. 30, 2022, as Appl. No. 18/091,687.
Application 18/091,687 is a division of application No. 17/043,339, granted, now 11,572,551, previously published as PCT/JP2019/013926, filed on Mar. 29, 2019.
Claims priority of application No. 2018-070188 (JP), filed on Mar. 30, 2018.
Prior Publication US 2023/0323317 A1, Oct. 12, 2023
Int. Cl. C12N 9/18 (2006.01); C12P 7/56 (2006.01); C12P 7/62 (2022.01); C12P 41/00 (2006.01)
CPC C12N 9/18 (2013.01) [C12P 7/62 (2013.01); C12P 41/00 (2013.01); C12Y 301/01 (2013.01)] 7 Claims
OG exemplary drawing
 
5. 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):

OG Complex Work Unit Chemistry
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.