US 2023/0203456 A1Published application

L-glutamate oxidase mutant

AssigneeAjinomoto Co., Inc.

InventorsKazutoshi Takahashi, Hiroki Yamaguchi, Uno Tagami

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Abstract

The present invention provides an alternative L-glutamate oxidase that allows for measurement of L-glutamate. More specifically, the present invention provides the following L-glutamate oxidase mutant (a) or (b) and the like: (a) an L-glutamate oxidase mutant including an amino acid sequence that has 90% or more identity to an amino acid sequence of SEQ ID NO: 3 and exhibits an activity of oxidizing L-glutamate, except an L-glutamate oxidase including an amino acid sequence of SEQ ID NO: 1; or (b) an L-glutamate oxidase mutant comprising a peptide linker consisting of 1 to 20 amino acid residues which is inserted into one or more sites selected from the group consisting of (1) a site in a region proximity to a boundary between α1 and α2 regions, (2) a site in a region proximity to a boundary between α2 and γ regions and (3) a site in a region proximity to a boundary between γ and β regions in the L-glutamate oxidase mutant (a), and having the activity of oxidizing L-glutamate.

Claims19

Dependent claims

An L-glutamate oxidase mutant of (a) or (b):

(a) an L-glutamate oxidase mutant of the protein having the amino acid sequence of SE ID NO:
1, wherein said mutant comprises an amino acid sequence that has 90% or more identity to the amino acid sequence of SEQ ID NO:
3 and exhibits an activity of oxidizing L-glutamate,;
or
(b) an L-glutamate oxidase mutant able to oxidize L-glutamate, wherein said mutant comprises a peptide linker consisting of 1 to 20 amino acid residues which is inserted into a site selected from the group consisting of:
(1) a site in a region in proximity to a boundary between α1 and α2 regions, wherein the site consists of the 349th to 363rd amino acid residues of SEQ ID NO:
3,
(2) a site in a region in proximity to a boundary between α2 and γ regions, wherein the site consists of the 372nd to 377th amino acid residues of SEQ ID NO:
3,
(3) a site in a region in proximity to a boundary between γ and β regions, wherein the site consists of the 466th to 469th amino acid amino acid residues of SEQ ID NO:
3, and
(4) combinations thereof.

9 more elements

Dependencies are inferred from the claim text.

Facts

Application: 17/947,596Filed 19 Sep 2022
Priority: JP 2020-05280924 Mar 2020
Published: 29 Jun 2023
Term ends: Not estimatedThe source records don’t support an estimate.

Classification13 codes

C12N 9/0022
Acting on the CH-NH2 group of donors: with oxygen as acceptor
C12Q 1/26
Measuring or testing processes involving enzymes, nucleic acids or microorganisms ; Compositions therefor; Processes of preparing such…
C12N 15/70
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression: vectors or expression systems…
C12P 7/50
Polycarboxylic acids: having keto groups, e.g. 2-ketoglutaric acid