Variants of Erwinia-type creatinase
AssigneeRoche Diagnostics Operations, Inc.
InventorsZhixin Shao, Rainer Schmuck, Peter Kratzsch, Janet Kenklies, Harald Weisser
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Abstract
The present invention relates to mutated genes encoding variants of an Erwinia-type creatinase (EC 3.5.3.3, alternative name: creatine amidinohydrolase), to the creatinase variants encoded by these genes, and to different applications of these variants of creatinase, particularly to their use for determining the creatinine and/or creatine concentration in a sample.
Claims12
A variant of an erwinia-type creatinase modified relative to a wild-type creatinase having creatinase activity, characterized in that said variant comprises at least one amino acid substitutions at a position corresponding to a position of the amino acid sequence shown in SEQ ID NO:
- 2 selected from the group of positions consisting of N130, M203, I278, I304, and F395.
A reagent for determination of creatine comprising an Erwinia-type creatinase variant according to any of the preceding claims.
Method for production of an Erwinia-type creatinase variant, the method comprising the steps of cloning a wild-type enzyme, introducing mutations, selecting as compared to the wild-type enzyme variants with improved thermostability or improved creatinase activity, and recombinantly producing said variant.
Dependencies are inferred from the claim text.