Modified aminotransferase, gene thereof, and method for producing optically active amino compound using same
AssigneeKaneka Corporation
InventorsNoriyuki Ito, Shigeru Kawano, Yoshihiko Yasohara
Life of this patent
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Abstract
Provided is a method for efficiently producing from a ketone compound an optically active amino compound useful as an intermediate of a drug, an agricultural chemical, or the like. Provided are: a polypeptide having aminotransferase activity that is increased in stereoselectivity, heat resistance, and resistance to amine compounds compared to the wild type enzyme by means of modifying an aminotransferase derived from Pseudomonas fluorescens; a gene encoding the polypeptide; and a transformant that expresses the gene at a high level.
Claims20
A polypeptide having the following properties (i) to (iii):
- (i) having at least 85% sequence identity to an amino acid sequence of SEQ ID NO:
- 1;
- (ii) being capable of catalyzing a reaction of 7-methoxy-2-tetralone to produce (S)-7-methoxy-2-aminotetralin in the presence of an amino group donor;
- and
- (iii) having reactivity higher than that of an aminotransferase consisting of the amino acid sequence of SEQ ID NO:
- 1.
A polypeptide selected from the group consisting of the following (A) to (C):
- (A) a polypeptide that consists of an amino acid sequence in which one or more amino acids have been substituted at one or more positions selected from positions 161, 420, 17, 84, 171, 176, 262, 302, 421, 435, 29, 42, 116, 153, 190, 284, 209, 235, 236, 408, 418, 434, 442, 3, 11, and 151 in the amino acid sequence of SEQ ID NO:
- 1;
- (B) a polypeptide that consists of an amino acid sequence in which one or more amino acids have been substituted at one or more positions selected from positions 161, 420, 17, 84, 171, 176, 262, 302, 421, 435, 29, 42, 116, 153, 190, 284, 209, 235, 236, 408, 418, 434, 442, 3, 11, and 151, and one or more amino acids at one or more positions other than the above amino acid positions has been substituted, added, inserted, or deleted in the amino acid sequence of SEQ ID NO:
- 1, and that is capable of catalyzing a reaction of 7-methoxy-2-tetralone to produce (S)-7-methoxy-2-aminotetralin in the presence of an amino group donor, and has reactivity higher than that of an aminotransferase consisting of the amino acid sequence of SEQ ID NO:
- 1;
- and
- (C) a polypeptide that consists of an amino acid sequence in which one or more amino acids have been substituted at one or more positions selected from positions 161, 420, 17, 84, 171, 176, 262, 302, 421, 435, 29, 42, 116, 153, 190, 284, 209, 235, 236, 408, 418, 434, 442, 3, 11, and 151 in the amino acid sequence of SEQ ID NO:
- 1, and a sequence identity to the amino acid sequence of SEQ ID NO:
- 1, except for the above substituted position or positions, is at least 85%, and that is capable of catalyzing a reaction of 7-methoxy-2-tetralone to produce (S)-7-methoxy-2-aminotetralin in the presence of an amino group donor, and has reactivity higher than that of an aminotransferase consisting of the amino acid sequence of SEQ ID NO:
- 1.
Dependencies are inferred from the claim text.