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US 2002/0173013 A1Published application
Esterace genes and uses of the same
AssigneeSumitomo Chemical Company, Limited
InventorsKae Kishimoto, Yuko Kobayashi, Yoshiki Takashima, Ayumu Inoue
Life of this patent
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Abstract
The present invention provides a novel gene encoding a protein having an excellent catalyst ability for producing (S)-N-substituted cyclic imino acid represented by the general formula (2) (=the (S)-cyclic imino acid (2)): 1 and to provide a novel method for producing the (S)-cyclic imino acid (2) by an asymmetric hydrolization of the N-substituted cyclic imino acid ester represented by the general formula (1): 2 in a manner of gene engineering technology utilizing the novel gene provided.
Claims16
A gene comprising any one of the following nucleotide sequences:
- (a) the nucleotide sequence represented by SEQ ID NO:2;
- (b) a nucleotide sequence which encodes an amino acid sequence of a protein capable of hydrolyzing a racemic ethyl N-benzylazetidine-2-carboxylate asymmetrically and producing an (S)-N-benzylazetidine-2-carboxylic acid predominantly and which consists of a part of the nucleotide sequence represented by SEQ ID NO:2;
- (c) a nucleotide sequence which encodes an amino acid sequence of a protein capable of hydrolyzing a racemic ethyl N-benzylazetidine-2-carboxylate asymmetrically and producing an (S)-N-benzylazetidine-2-carboxylic acid predominantly and which hybridizes under a stringent condition with a DNA consisting of a nucleotide sequence encoding the amino acid sequence represented by SEQ ID NO:
- 1;
- (d) a nucleotide sequence which encodes an amino acid sequence of a protein capable of hydrolyzing a racemic ethyl N-benzylazetidine-2-carboxylate asymmetrically and producing an (S)-N-benzylazetidine-2-carboxylic acid predominantly and which encodes an amino acid sequence of a protein having a molecular weight of about 25,000 daltons determined by an SDS-PAGE;
- (e) a nucleotide sequence which encodes an amino acid sequence of a protein capable of hydrolyzing a racemic ethyl N-benzylazetidine-2-carboxylate asymmetrically and producing an (S)-N-benzylazetidine-2-carboxylic acid predominantly and which has a nucleotide homology of 60% or higher with a nucleotide sequence represented by Nucleotide No.235 to 765 in the nucleotide sequence represented by SEQ ID NO:2;
- and,
- (f) a nucleotide sequence of a DNA which is amplified by a PCR using as a primer an oligonucleotide allowing a DNA having a nucleotide sequence represented by Nucleotide No.235 to 765 in the nucleotide sequence represented by SEQ ID NO:2 to be capable of being amplified and as a template a chromosome DNA derived from a microorganism belong to Aspergillus flavus.
5 more elements
A gene comprising any one of the following nucleotide sequences:
- (a) the nucleotide sequence represented by SEQ ID NO:2;
- (b) a nucleotide sequence represented by Nucleotide No.103 to 765 in the nucleotide sequence represented by SEQ ID NO:2;
- (c) a nucleotide sequence represented by Nucleotide No.235 to 765 in the nucleotide sequence represented by SEQ ID NO:2;
- (d) a nucleotide sequence encoding the amino acid sequence represented by SEQ ID NO:1;
- (e) a nucleotide sequence encoding an amino acid sequence represented by Amino Acid No.35 to 255 in the amino acid sequence represented by SEQ ID NO:
- 1;
- and,
- (f) a nucleotide sequence encoding an amino acid sequence represented by Amino Acid No.79 to 255 in the amino acid sequence represented by SEQ ID NO:
- 1.
6 more elements
A protein comprising any one of the following amino acid sequences:
- (a) the amino acid sequence represented by SEQ ID NO:1;
- (b) an amino acid sequence of a protein which is capable of hydrolyzing a racemic ethyl N-benzylazetidine-2-carboxylate asymmetrically and producing an (S)-N-benzylazetidine-2-carboxylic acid predominantly and which consists of a part of the amino acid sequence represented by SEQ ID NO:1;
- (c) an amino acid sequence of a protein which is capable of hydrolyzing a racemic ethyl N-benzylazetidine-2-carboxylate asymmetrically and producing an (S)-N-benzylazetidine-2-carboxylic acid predominantly and which is encoded by a nucleotide sequence which hybridizes under a stringent condition with a DNA consisting of a nucleotide sequence encoding the amino acid sequence represented by SEQ ID NO:1;
- (d) an amino acid sequence of a protein which is capable of hydrolyzing a racemic ethyl N-benzylazetidine-2-carboxylate asymmetrically and producing an (S)-N-benzylazetidine-2-carboxylic acid predominantly and which has a molecular weight of about 25,000 daltons determined by an SDS-PAGE;
- and,
- (e) an amino acid sequence of a protein which is capable of hydrolyzing a racemic ethyl N-benzylazetidine-2-carboxylate asymmetrically and producing an (S)-N-benzylazetidine-2-carboxylic acid predominantly and which has an amino acid homology of 60% or higher with the amino acid sequence represented by SEQ ID NO:1.
3 more elements
A protein comprising any one of the following amino acid sequences:
- (a) the amino acid sequence represented by SEQ ID NO:1;
- (b) an amino acid sequence represented by Amino Acid No.35 to 255 in the amino acid sequence represented by SEQ ID NO:1;
- (c) an amino acid sequence represented by Amino Acid No.79 to 255 in the amino acid sequence represented by SEQ ID NO:1;
- (d) an amino acid sequence corresponding to the nucleotide sequence represented by SEQ ID NO:2;
- (e) an amino acid sequence corresponding to the nucleotide sequence represented by Nucleotide No.103 to 765 in the nucleotide sequence represented by SEQ ID NO:2;
- and,
- (f) an amino acid sequence corresponding to the nucleotide sequence represented by Nucleotide No.235 to 765 in the nucleotide sequence represented by SEQ ID NO:2.
4 more elements
Dependencies are inferred from the claim text.