Alteration of substrate specificity of fructosyl-amino acid oxidase from Ulocladium sp. JS-103.
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).
Fujiwara
Maki
M
Department of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Osaka, Japan.
Sumitani
Jun-ichi
J
Koga
Shinji
S
Yoshioka
Issei
I
Kouzuma
Takuji
T
Imamura
Shigeyuki
S
Kawaguchi
Takashi
T
Arai
Motoo
M
D000594:Amino Acid Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D019943:Amino Acid Substitution; D001203:Ascomycota / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*; D001681:Biological Assay / Q000379:methods; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006442:Glycated Hemoglobin / Q000032:analysis; D006801:Humans; D016296:Mutagenesis / Q000235:genetics*; D020125:Mutation, Missense; D013379:Substrate Specificity / Q000235:genetics; D014633:Valine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism
D000594:Amino Acid Oxidoreductases; D005656:Fungal Proteins; D006442:Glycated Hemoglobin; D014633:Valine; C045420:fructosylvaline; C103012:amadoriase