著者

Khondkar Ehteshamul KABIR Daizo HIROWATARI Katsuhiro WATANABE Daizo KOGA

出版者

Japan Society for Bioscience, Biotechnology, and Agrochemistry

雑誌

Bioscience, Biotechnology, and Biochemistry (ISSN:09168451)

巻号頁・発行日

vol.70, no.1, pp.252-262, 2006 (Released:2006-01-23)

参考文献数

60

被引用文献数

17

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75-kDa chitinase, which showed potential as a biocontrol agent against Japanese pine sawyer, was characterized after purification from the integument of the fifth instar larvae of Bombyx mori by chromatography on diethylaminoethyl (DEAE)-Toyoperal 650 (M), hydroxylapatite, and Fractogel EMD DEAE 650 (M) columns. The optimum pH was 6.0 toward N-acetylchitopentaose (GlcNAc5) and 10 toward glycolchitin. The optimum temperature was 60 °C toward GlcNAc5 and 25 °C toward glycolchitn. The enzyme was stable at pH 7–10 and below 40 °C. Kinetic analysis and reaction-pattern analysis using glycolchitin and N-acetylchitooligosacchraides as substrates indicated that 75-kDa chitinase is an endo- or random-type hydrolytic enzyme to produce the β anomeric product and that it prefers the longer N-acetylchitooligosaccharides, suggesting, together with the N-terminal amino acid sequence, that the 75-kDa chitinase belongs to family 18 of glycosyl hydrolases.