ID | 64346 |
Sort Key | 5
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Title Alternative | Studies on l-Glutamate Oxidase with Strict Substrate Specificity from Streptomyces sp.
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FullText URL | |
Author |
Nakayama, Natsume
Course of Agrochemical Bioscience
Inagaki, K.
Kaken ID
researchmap
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Abstract | l-glutamate oxidase (LGOX) from Streptomyces sp. is a heterohexameric flavin enzyme that catalyzes the oxidative deamination of l-glutamate to form α-ketoglutarate with ammonia and hydrogen peroxide. LGOX shows strict substrate specificity for l-Glu. In addition, it is highly thermostable and pH stable. Because of these properties, LGOX is currently used as a biosensor for the trace determination of l-Glu in the food industry and clinical laboratories. The full-length cDNA is 2103 bp and is encoded by a single polypeptide chain consisting of 701 residues including subunits α-γ-β. The LGOX gene was heterologously expressed in Escherichia coli JM109. The LGOX precursor expressed in E. coli is a homodimer with weak enzymatic activity and becomes a heterohexamer upon activation by protease treatment. X-ray crystallography and docking studies of purified recombinant LGOX suggest that the Arg305 residue is a key residue for substrate recognition. Mutant analysis showed that Arg305 is essential for substrate recognition, as the activity toward l-Glu was greatly reduced and substrate specificity was changed in some enzymes. The functional analysis of R305E-LGOX, which is an l-Arg oxidase, revealed that R305E-LGOX can be used as a enzyme biosensor for l-Arg.
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Keywords | l-glutamate oxidase
biosensor
substrate recognition
X-ray crystallography
modification of substrate specificity
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Note | 総合論文 (Comprehensive paper)
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Publication Title |
Scientific Reports of the Faculty of Agriculture, Okayama University
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Published Date | 2023-02-01
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Volume | volume112
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Publisher | 岡山大学農学部
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Publisher Alternative | The Faculty of Agriculture, Okayama University
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Start Page | 13
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End Page | 18
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ISSN | 2186-7755
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Content Type |
Departmental Bulletin Paper
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OAI-PMH Set |
岡山大学
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language |
Japanese
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File Version | publisher
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Eprints Journal Name | srfa
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