Modified glycine oxidase
AssigneeAjinomoto Co., Inc.
InventorsYuya Kodama, Wataru Hoshino, Kazutoshi Takahashi, Moemi Ito
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Abstract
The present invention provides a novel enzyme and methods of using the enzyme for measuring glycine concentration. Specifically, the present invention provides an enzyme in which at least one amino acid residue is mutated so as to improve a property of a glycine oxidase which is associated with the measurement of glycine (e.g., activity of glycine oxidase for glycine, thermal stability of glycine oxidase, and substrate specificity of glycine oxidase for glycine,); and a method of analyzing glycine, that includes measuring glycine contained in a test sample using the modified enzyme; and the like.
Claims13
A glycine oxidase enzyme comprising the amino acid sequence of SEQ ID No:
- 2, and also comprising a mutation of at least one amino acid residue, wherein said mutation results in the improvement of one or more properties of said glycine oxidase which are associated with measurement of glycine as compared to said glycine oxidase that does not comprise said mutation, wherein the one or more properties are selected from the group consisting of:
- (a) an activity of the glycine oxidase for glycine;
- (b) a thermal stability of the glycine oxidase;
- and
- (c) a substrate specificity of the glycine oxidase for glycine,
- wherein the mutation is selected from the group consisting of:
- (A) a substitution of the first threonine in a TTS motif with alanine, cysteine, or glycine,
- (B) a substitution of serine in the TTS motif with lysine,
- (C) a substitution of cysteine in an HCY motif with alanine, aspartic acid, glycine,
- histidine, asparagine, tryptophan, tyrosine, or serine,
- (D) a substitution of leucine in an LRP motif with isoleucine, valine, cysteine, threonine, or proline,
- (E) a substitution of serine with arginine in a SG motif located in at positions 190 to 204 of SEQ ID NO:
- 2,
- (F) a substitution of glycine in a POT motif with tyrosine or glutamine, and
- (G) combinations thereof;
- wherein the glycine oxidase enzyme may have supplemental mutations as long as said supplemental mutations maintain said improved property.
Dependencies are inferred from the claim text.