A Chitinase Indispensable for Formation of Protoplast of Schizophyllum commune in Basidiomycete-lytic Enzyme Preparation Produced by Bacillus circulans KA-304
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概要
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KA-prep, a culture filtrate of Bacillus circulans KA-304 grown on a cell-wall preparation of Schizophyllum commune, has an activity to form protoplasts from S. commune mycelia. α-1,3-Glucanase, which was isolated from an ammonium sulfate fraction of 0–30% saturation of KA-prep, gave the protoplast-forming activity to an ammonium sulfate fraction of 30–50% saturation of KA-prep, which contained chitinase(s) and β-glucanase(s) but was inactive in the protoplast formation. Chitinase(s) and β-glucanase(s) in the ammonium sulfate fraction of 30–50% saturation were separated by DEAE-cellulofine A-500 column chromatography, and the protoplast-forming activity appeared when the chitinase preparation was mixed with the α-1,3-glucanase. The β-glucanase preparation was not effective for the protoplast formation whereas its addition enhanced the protoplast-forming activity of the mixture of α-1,3-glucanase and the chitinase preparation. The chitinase preparation contained two chitinases (chitinase I and II). Chitinase I showed the protoplast-forming activity with α-1,3-glucanase, but chitinase II did not. Chitinase I, a monomeric protein with a molecular weight of 41,000, was active toward colloidal chitin and ethylene glycol chitin. Chitinase I produced predominantly N,N′-diacetylchitobiose and N,N′,N″-triacetylchitotriose from colloidal chitin, and the enzyme was inactive to p-NP-β-D-N-acetylglucosaminide, suggesting that it was an endo-type enzyme. The N-terminal amino acid sequence of chitinase I (A L A T P T L N V S A S S G M) had no sequential identity to those of known chitinases.
- 社団法人 日本農芸化学会の論文
- 2004-06-23
著者
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TACHIKI TAKASHI
Department of Food Science and Technology, Kyoto University
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YANO Shigekazu
Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University
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WAKAYAMA Mamoru
Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University
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Wakayama Mamoru
Dep. Of Biotechnology Fac. Of Life Sciences Ritsumeikan Univ.
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Wakayama Mamoru
Department Of Bioscience And Biotechnology Faculty Of Science And Engineering Ritsumeikan University
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Rattanakit Nopakarn
Department of Chemistry, Faculty of Science, Chiang Mai University
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Tachiki T
Department Of Food Science And Technology Faculty Of Agriculture Kyoto University
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Tachiki Takashi
Department Of Bioscience & Technology Faculty Of Science & Engineering Ritsumeikan Universit
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Yano Shigekazu
Department Of Biotechnology College Of Life Sciences Ritsumeikan University
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Rattanakit Nopakarn
Department Of Chemistry Faculty Of Science Chiang Mai University
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Yano Shuntaro
Department Of Biotechnology Faculty Of Life Sciences Ritsumeikan University
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Wakayama Mamoru
Department Of Biotechnology Faculty Of Life Sciences Ritsumeikan University
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YANO Shigekazu
Department of Bioscience and Biotechnology, Faculty of Science and Engineering, Ritsumeikan University
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Wakayama Mamoru
Department of Applied Chemistry, Faculty of Engineering, Oita University
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