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US 11,142,755 B2Granted
Mutant glutathione synthetase and method for producing gamma-glutamyl-valyl-glycine
AssigneeAjinomoto Co., Inc.
InventorsAyako Sato, Eri Higashiura, Misato Okamoto, Takayuki Ito, Erika Watanabe, Uno Tagami, Yuki Oda, Tatsuki Kashiwagi, Masayuki Sugiki
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Abstract
A mutant glutathione synthetase (GSHB) suitable for generating γ-Glu-Val-Gly, and a method for producing γ-Glu-Val-Gly using the same are provided. γ-Glu-Val-Gly is produced by using a mutant GSHB having a mutation at such a position as V7, N13, I14, N15, K17, F22, F95, M165, N199, Y200, P202, I274, T285, and P287.
Claims22
A method for producing γ-Glu-Val-Gly and/or a salt thereof, the method comprising:
- reacting a mutant glutathione synthetase with γ-Glu-Val and Gly to generate γ-Glu-Val-Gly,
- wherein the mutant glutathione synthetase has γ-glutamylvalylglycine synthetase activity and comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:
- 2, where at least one amino acid residue in the mutant glutathione synthetase corresponding to the following amino acid residue (A) or (B) is mutated to a different amino acid:
- (A) at least one amino acid residue selected from the group consisting of F22, M165, I274, T285, and P287 in SEQ ID NO:
- 2;
- (B) a combination of at least one amino acid residue selected from the group consisting of V7, N13, I14, N15, K17, F95, N199, Y200, and P202 in SEQ ID NO:2 and the amino acid residue (A).
3 more elements
A method for producing γ-Glu-Val-Gly and/or a salt thereof, the method comprising:
- reacting a γ-glutamylvaline synthetase and a mutant glutathione synthetase with Glu, Val, and Gly to generate γ-Glu-Val-Gly,
- wherein the mutant glutathione synthetase has γ-glutamylvalylglycine synthetase activity and comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:
- 2, where at least one amino acid residue in the mutant glutathione synthetase corresponding to the following amino acid residue (A) or (B) is mutated to a different amino acid:
- (A) at least one amino acid residue selected from the group consisting of F22, M165, I274, T285, and P287 in ID NO:
- 2;
- (B) a combination of at least one amino acid residue selected from the group consisting of V7, N13, I14, N15, K17, F95, N199, Y200, and P202 in SEQ ID NO:
- 2 and the amino acid residue (A).
4 more elements
A method for producing γ-Glu-Val-Gly and/or a salt thereof, the method comprising
- culturing a microorganism having a mutant glutathione synthetase in a culture medium to generate γ-Glu-Val-Gly from γ-Glu-Val and Gly,
- wherein the mutant glutathione synthetase has γ-glutamylvalylglycine synthetase activity and comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:
- 2, where at least one amino acid residue in the mutant glutathione synthetase corresponding to the following amino acid residue (A) or (B) is mutated to a different amino acid:
- (A) at least one amino acid residue selected from the group consisting of F22, M165, I274, T285, and P287 in SEQ ID NO:
- 2;
- (B) a combination of at least one amino acid residue selected from the group consisting of V7, N13, I14, N15, K17, F95, N199, Y200, and P202 in SEQ ID NO:
- 2 and the amino acid residue (A).
4 more elements
A method for producing γ-Glu-Val-Gly and/or a salt thereof, the method comprising:
- culturing a microorganism in a culture medium to generate γ-Glu-Val-Gly from Glu, Val, and Gly, where the microorganism comprises γ-glutamylvaline synthetase and a mutant glutathione synthetase,
- wherein the mutant glutathione synthetase has γ-glutamylvalylglycine synthetase activity and comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:
- 2, where at least one amino acid residue in the mutant glutathione synthetase corresponding to the following amino acid residue (A) or (B) is mutated to a different amino acid:
- (A) at least one amino acid residue selected from the group consisting of F22, M165, I274, T285, and P287 in SEQ ID NO:
- 2;
- (B) a combination of at least one amino acid residue selected from the group consisting of V7, N13, I14, N15, K17, F95, N199, Y200, and P202 in SEQ ID NO:
- 2 and the amino acid residue (A).
4 more elements
Dependencies are inferred from the claim text.