Scientific Reports of the Faculty of Agriculture, Okayama University
Published by the Faculty of Agriculture, Okayama University
ONLINE ISSN : 2186-7755


Nakai, Ryuichiro
Fujino, Shihoko
Utsumi, Tomohiro
Kusakabea, Hitoshi
Inagaki, Kenji Kaken ID researchmap
 L‒Glutamate oxidase (LGOX) from Streptomyces sp. X‒119‒6 has strict substrate specificity toward L‒glutamate. Recently, we solved the X‒ray crystal structure of LGOX and this revealed that Arg305 in the active site is the key residue involved in substrate recognition. Therefore, we created 19 mutant enzymes of R305X‒LGOX by saturation mutagenesis. One of them R305D‒LGOX, Arg305 substituted with Asp exhibited oxidase activity for L‒Arg. Optimum pH of R305D‒LGOX mutant enzyme was pH 8.5. Interestingly, the activity of R305D‒LGOX toward L‒Arg was inhibited by phosphate. And furthermore, the substrate specificity of R305D‒LGOX was affected by using buffer. The results of inhibition analysis suggest, that phosphate is a competitive inhibitor of R305D‒LGOX when L‒Arg is used as substrate. Kinetic analysis of R305D‒LGOX showed that Km value and kcat value of R305D‒LGOX toward l-Arg were 0.68 mM and 6.7 s-1 respectively. In this study, we showed that R305D‒LGOX mutant enzyme is a novel l-arginine oxidase and useful for l-arginine biosensor.
L-glutamate oxidase
L-arginine oxidase
modified substrate specificity
L-amino acid oxidase
原著論文 (Original paper)