Novel glutathione oxidase, its production and use
AssigneeTakara Suzo Co., Ltd.
InventorsSusumu Matsui, Satoko Uchida, Tsutomu Taniguchi
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Abstract
A novel glutathione oxidase which acts on reduced glutathione alone and not no other sulfhydryl compounds is obtained by culturing Calodon or Cortinarius of the class Basidiomycetes.
Claims7
A novel glutathione oxidase T-1 having the following physicochemical properties:
- (1) Action:
- Acts on reduced glutathione in the presence of oxygen to produce oxidized glutathione and hydrogen peroxide;
- (2) Substrate specificity:
- Acts on reduced glutathione alone, not on other sulfhydryl compounds at all;
- (3) Optimum pH and pH stability:
- Has an optimum pH in the vicinity of 7.0, and is stable between pH 4.0 and pH 8.0 for 60-minute treatment at 37.degree. C.;
- (4) Optimum temperature and thermal stability:
- Has an optimum temperature in the vicinity of 50.degree. C., and is stable up to 60.degree. C. for 10-minute treatment at a pH of 7.0;
- (5) Molecular weight:
- About 450000 by the gel filtration method, and about 55000 by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, from which said novel glutathione oxidase T-1 is composed of eight subunits;
- (6) Coenzyme:
- Contains flavine adenine dinucleotide (FAD), a coenzyme, of 4 molecules/molecule;
- (7) Isoelectric point:
- 5.65.+-.0.05;
- (8) Interfering agent:
- Hg.sup.2+, p-chloromercuribenzoic acid, sodium L-ascorbate, etc;
- (9) Sugar content:
- About 40%.
A method for the quantitative determination of reduced glutathione characterized in that reduced glutathione in test samples is oxidized with said novel glutathione oxidase T-1 in the presence of oxygen, and the reduced glutathione is quantitatively determined based on the amount of consumed oxygen or formed hydrogen peroxide in the reaction solution.
A method for producing said novel glutathione oxidase T-1 characterized in that a novel glutathione oxidase T-1 producing strain belonging to the genus Calodon or Cortinarius is cultured and the novel glutathione oxidase T-1 produced is collected from the cultured broth obtained.
Dependencies are inferred from the claim text.