Esterase gene and its use
AssigneeSumitomo Chemical Company, Limited
InventorsTakeshi Ishii, Satoshi Mitsuda
Life of this patent
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Abstract
An isolated esterase gene coding for an esterase capable of causing asymmetric hydrolysis of an organic carboxylic acid ester 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, to produce the cyclopentenolone of formula I in (R)-form, and hybridizing to the base sequence of SEQ ID NO:1, is useful for the industrially favorable production of optically active cyclopentenolones of formula I which serve as the intermediates of drugs, agricultural chemicals or other active products.
Claims8
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 6×SSC (standard saline citrate), 0.5% (w/v) SDS (sodium dodecyl sulfate), 100 μg/ml salmon sperm DNA, 63° 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 6×SSC (standard saline citrate), 0.5% (w/v) SDS (sodium dodecyl sulfate), 100 μg/ml salmon sperm DNA, 63° C.
Dependencies are inferred from the claim text.