Patent ID: 6472189
Filing Date: 2002-10-29
Classification: C12N,C12P

Abstract:
A method for the optical resolution of a cyclopentenolone of Formula I: wherein R1 is hydrogen or methyl, and R2 is C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C1-C4 haloalkyl, a C5-C9 aliphatic hydrocarbon moiety which may be optionally protected on the terminal hydroxyl group thereof, or a C5-C9 fatty acid residue which may be optionally protected on the terminal carboxyl group thereof, said optical resolution method comprising:(I) allowing an esterase to act upon an organic carboxylic acid ester of the cyclopentenolone of formula I for asymmetric hydrolysis of the ester; and (II) separating the cyclopentenolone of formula I in (R)-form from the ester of the corresponding enantiomer thereof, wherein said esterase is selected from the group consisting of: (1) an esterase according to the amino acid sequence of SEQ ID NO: 2, (2) an esterase having 363 amino acids and being coded for by an isolated polynucleotide, said isolated polynucleotide having: (a) a recognition sequence for the restriction endonuclease for SalI at a point equivalent to nucleotides 537-542 of SEQ ID NO: 1 in said isolated polynucleotide, (b) no recognition sequence for the restriction endonuclease SmaI and XhoI, and (c) a full complement of a nucleotide sequence hybridizing to SEQ ID NO: 1 under conditions of 6Ã—SSC (standard saline citrate), 0.5% (w/v) SDS (sodium dodecyl sulfate), 100 &mgr;g/ml salmon sperm DNA, 63Â° C.; (3) an esterase that is coded by an isolated polynucleotide, said isolated polynucleotide being contained in the deposited strain FERM BP-5740, (4) an esterase that is coded by an isolated polynucleotide, said isolated polynucleotide having a base sequence of SEQ ID NO:1 and (5) an esterase that is coded by an isolated polynucleotide, said isolated polynucleotide coding for a region in SEQ ID NO:2 of which N-terminal amino acid sequence is the same as defined in SEQ ID NO:3 which is identical to residues 44-58 of SEQ ID NO:2 and is a full complement to a nucleotide sequence hybridizing to SEQ ID NO:1 under conditions of 6Ã—SSC (standard saline citrate), 0.5% (w/v) SDS (sodium dodecyl sulfate), 100 &mgr;g/ml salmon sperm DNA, 63Â° C.