Mutant glutamate-cysteine ligase and method for manufacturing gamma glutamyl-valyl-glycine
AssigneeAjinomoto Co., Inc.
InventorsAyako Sasahara, Eri Tabuchi, Hideyuki Suzuki, Uno Tagami, Tatsuki Kashiwagi, Takayuki Ito, Hiroyuki Nozaki, Shunichi Suzuki
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Abstract
A mutant glutamate-cysteine ligase (GSHA) suitable for generating γ-Glu-Val, and a method for producing γ-Glu-Val-Gly using the same are provided. γ-Glu-Val is produced by using a mutant GSHA having a specific mutation with Glu and Val as raw materials, and γ-Glu-Val-Gly is further produced by using γ-Glu-Val and Gly as raw materials. γ-Glu-Val-Gly is produced by using a mutant GSHA having a specific mutation with Glu, Val, and Gly as raw materials.
Claims23
A method for producing γ-Glu-Val and/or a salt thereof, the method comprising:
- contacting a mutant glutamate-cysteine ligase with Glu and Val to produce γ-Glu-Val and/or a salt thereof,
- wherein the mutant glutamate-cysteine ligase comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:
- 2 wherein the amino acid residue corresponding to at least one residue selected from the group consisting of L135, Q144, Y241, N243, and Y300 of SEQ ID NO:
- 2 is mutated to a different amino acid in the mutant glutamate-cysteine ligase, and
- wherein the mutant has γ-glutamylvaline synthetase activity.
A method for producing γ-Glu-Val-Gly and/or a salt thereof, the method comprising:
- contacting a mutant glutamate-cysteine ligase with Glu and Val to produce γ-Glu-Val;
- contacting a glutathione synthetase with the produced γ-Glu-Val and Gly to produce γ-Glu-Val-Gly and/or a salt thereof,
- wherein the mutant glutamate-cysteine ligase comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:
- 2 wherein the amino acid residue corresponding to at least one residue selected from the group consisting of L135, Q144, Y241, N243, and Y300 of SEQ ID NO:
- 2 is mutated to a different amino acid in the mutant glutamate-cysteine ligase, and
- wherein the mutant has γ-glutamylvaline synthetase activity.
A method for producing γ-Glu-Val-Gly and/or a salt thereof, the method comprising:
- contacting a mutant glutamate-cysteine ligase and a glutathione synthetase with Glu, Val, and Gly to produce γ-Glu-Val-Gly and/or a salt thereof,
- wherein the mutant glutamate-cysteine ligase comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:
- 2 wherein the amino acid residue corresponding to at least one residue selected from the group consisting of L135, Q144, Y241, N243, and Y300 of SEQ ID NO:
- 2 is mutated to a different amino acid in the mutant glutamate-cysteine ligase, and
- wherein the mutant has γ-glutamylvaline synthetase activity.
Dependencies are inferred from the claim text.