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US 6,410,253 B1Granted
Cholesterol oxidase
AssigneeKyowa Hakko Kogyo Co., Ltd.
InventorsKinya Fujishiro, Takayuki Uwajima
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Abstract
A process for producing cholesterol oxidase from a recombinant microorganism, the microorganism itself and its recombinant plasmid are disclosed. The cholesterol oxidase has a particular amino acid sequence, a high substrate affinity and a working pH in the acidic range. The gene for this cholesterol oxidase was derived from Brevibacterium sterolicum. The cholesterol oxidase is used for the quantitative determination of cholesterol.
Claims4
A method for quantitative determination of cholesterol contained in a sample which comprises;
- exposing the sample to a cholesterol oxidase to oxidize the cholesterol, and
- measuring the amount of oxidized cholesterol or hydrogen peroxide thereby determining the amount of the cholesterol,
- wherein the cholesterol oxidase is coded by DNA which is hybridizable to DNA as defined in SEQ ID:
- NO:1, under stringent conditions, wherein said stringent conditions comprise hybridization at 45° C. in a hybridization buffer containing 53.0 g/l NaCl, 26.5 g/l sodium succinate, 10 mM EDTA, 5-fold Denhart solution, 0.5% SDS, 0.1 mg/ml thermally denatured bovine thymus-derived DNA for 16 hours and washing at 65° C. with 3-fold SSC.
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Dependencies are inferred from the claim text.