Abstract
We showed by random mutagenesis that one-amino-acid substitution at Arg94 in fructosyl-amino acid oxidase from Ulocladium sp. JS-103 enhanced substrate specificity toward fructosyl valine (FV), a model compound of hemoglobin A(1c). Kinetic analysis showed that the specificity of the R94W mutant enzyme toward FV was 14-fold that of the wild-type enzyme. The mutant enzyme obtained will be useful in developing an enzymatic measurement method for hemoglobin A(1c).
MeSH terms
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Amino Acid Oxidoreductases / chemistry
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Amino Acid Oxidoreductases / genetics*
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Amino Acid Oxidoreductases / metabolism
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Amino Acid Substitution*
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Ascomycota / chemistry
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Ascomycota / enzymology
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Ascomycota / genetics*
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Biological Assay / methods
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Fungal Proteins / chemistry
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Fungal Proteins / genetics*
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Fungal Proteins / metabolism
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Glycated Hemoglobin / analysis
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Humans
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Mutagenesis / genetics*
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Mutation, Missense*
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Substrate Specificity / genetics
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Valine / analogs & derivatives
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Valine / chemistry
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Valine / metabolism
Substances
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Fungal Proteins
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Glycated Hemoglobin A
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fructosylvaline
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Amino Acid Oxidoreductases
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amadoriase
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Valine