Mutated D-aminotransferase and method for producing optically active glutamic acid derivatives using the same
AssigneeAjinomoto Co., Inc.
InventorsMasakazu Sugiyama, Kunihiko Watanabe, Tatsuki Kashiwagi, Ei-ichiro Suzuki
Life of this patent
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Abstract
A D-aminotransferase can be modified so as to efficiently produce (2R, 4R)-monatin having high sweetness intensity from 4-(indol-3-ylmethyl)-4-hydroxy-2-oxoglutaric acid by substituting an amino acid at least at one of positions (positions 100, 180 to 183, 243 and 244) involved in efficiently producing the (2R, 4R)-monatin in an amino acid sequence of a wild-type D-aminotransferase represented in SEQ ID NO:2.
Claims38
An isolated DNA comprising a nucleotide sequence selected from the group consisting of:
- (A) the nucleotide sequence of SEQ ID NO:1, wherein said DNA encodes a protein having a D-aminotransferase activity;
- (B) a nucleotide sequence encoding a protein having D-aminotransferase activity which hybridizes under stringent conditions with another DNA composed of a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO:1, wherein said stringent conditions comprise 65° C., 0.1×SSC and 0.1% SDS;
- (C) a nucleotide sequence encoding a protein having the amino acid sequence of SEQ ID NO:2, wherein said protein has a D-aminotransferase activity;
- and
- (D) a nucleotide sequence encoding a protein having an amino acid sequence having substitution, deletion, insertion, and/or addition of one to ten amino acid residues in the amino acid sequence SEQ ID NO:2, wherein said protein has a D-aminotransferase activity.
An isolated DNA encoding a protein having an amino acid sequence selected from the following (A) and (B):
- (A) an amino acid sequence having substitution of an amino acid residue at a position selected from positions 100, 180 to 183, 243 and 244, or combinations thereof, in the amino acid sequence of SEQ ID NO:2;
- and
- (B) an amino acid sequence having substitution, deletion, insertion, and/or addition of one or several amino acid residues at position(s) other than positions 100, 180 to 183, 243 and 244 in the amino acid sequence (A), such that a total of one to ten amino acid residues of the amino acid sequence of SEQ ID NO:2 have been altered;
- wherein said protein has a D-aminotransferase activity, and wherein an amount of (2R, 4R)-monatin produced with said protein from 4-(indol-3-ylmethyl)-4-hydroxy-2-oxoglutaric acid is greater than that produced with a protein having the amino acid sequence of SEQ ID NO:2.
Dependencies are inferred from the claim text.