Creatine amidinohydrolase, production thereof and use thereof

a technology of amidinohydrolase and creatine, which is applied in the field of creatine amidinohydrolase, can solve the problems of lower heat stability and greater km value of creatine, enzymes are not suitable for clinical testing, and are produced from various known cell lines. , to achieve the effect of small km value, superior reactivity to creatine, and high sensitivity
USRE38687E1Inactive Publication Date: 2005-01-11TOYO TOYOBO CO LTD
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Patent Information

Authority / Receiving Office
US · United States
Current Assignee / Owner
TOYO TOYOBO CO LTD
Publication Date
2005-01-11
Estimated Expiration
Not applicable · inactive patent

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Abstract

A creatine amidinohydrolase having the following physicochemical properties: Action: catalyzing the following reaction; creatine + H2O -> sarcosine + urea Optimum temperature: about 40 - 50 DEG C Optimum pH: pH about 8.0 - 9.0 Heat stability: not more than about 50 DEG C (pH 7.5, 30 min) Km value for creatine in a coupling assay using a sarcosine oxidase and a peroxidase: about 3.5 - 10.0 mM Molecular weight: about 43,000 (SDS-PAGE) Isoelectric point: about 3.5, a method for producing said enzyme, comprising culture of microorganism producing said enzyme, a method for the determination of creatine or creatinine in a sample using said enzyme, and a reagent therefor. According to the present invention, a creatine amidinohydrolase having a smaller Km value than that of the conventionally known creatine amidinohydrolase can be produced in an industrially large amount, and can be used as a routine reagent for clinical tests for determining creatine and creatinine in biological samples.
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Description

.Iadd.CROSS-REFERENCE TO RELATED APPLICATIONSMore than one reissue application has been filed for the reissue of U.S. Pat. No. 6,080,553. The reissue applications are Application No. 09 / 940,941(the present application) and Application No. 10 / 807,228, which are divisional reissues of U.S. Pat. No. 6,080,553..Iaddend.FIELD OF THE INVENTIONThe present invention relates to a novel creatine amidinohydrolase, specifically, a novel creatine amidinohydrolase having a very low Km value for creatine, and a method for producing said enzyme. The present invention also relates to a method for the determination of creatine or creatinine in a sample by the use of said enzyme, and a reagent therefor.BACKGROUND OF THE INVENTIONA creatine and a creatinine are found in blood and urine. A quick and accurate determination of their amounts is very important in making diagnosis of the diseases such as uremia, chronic neophritis, acute neophritis, giantism, tonic muscular dystrophy and the like. For making...

Claims

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