ヨコヤマ　ケンジ   KENJI YOKOYAMA   横山　憲二    所属   応用生物学部 応用生物学科    職種   教授
言語種別	英語
発行・発表の年月	2017/01
形態種別	学術論文
査読	査読あり
標題	Identification and characterization of thermostable glucose dehydrogenases from thermophilic filamentous fungi
執筆形態	共著
掲載誌名	Applied Microbiology and Biotechnology
掲載区分	国外
出版社・発行元	Springer
巻・号・頁	101,pp.173-183
著者・共著者	zumichi Ozawa, Hisanori Iwasa, Noriko Sasaki, Nao Kinoshita, Atsunori Hiratsuka, Kenji Yokoyama
概要	FAD-dependent glucose dehydrogenase (FAD-GDH), which contains FAD as a cofactor, catalyzes the oxidation of D-glucose to D-glucono-1,5-lactone, and plays an important role in biosensors measuring blood glucose levels. In order to obtain a novel FAD-GDH gene homolog, we performed degenerate PCR screening of genomic DNAs from 17 species of thermophilic filamentous fungi. Two FAD-GDH gene homologs were identified and cloned from Talaromyces emersonii NBRC 31232 and Thermoascus crustaceus NBRC 9129. We then prepared the recombinant enzymes produced by Escherichia coli and Pichia pastoris. Absorption spectra and enzymatic assays revealed that the resulting enzymes contained oxidized FAD as a cofactor and exhibited glucose dehydrogenase activity. The transition midpoint temperatures (Tm) were 66.4°C and 62.5°C for glycosylated FAD-GDHs of Ta. emersonii and Th. crustaceus prepared by using P. pastoris as a host, respectively. Therefore, both FAD-GDHs exhibited high thermostability. In conclusion, we propose that these thermostable FAD-GDHs could be ideal enzymes for use as thermotolerant glucose sensors with high accuracy.
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