Family 19 Chitinases from Streptomyces thermoviolaceus OPC-520 : Molecular Cloning and Characterization
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概要
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Family 19 chitinase genes, chi35 and chi25 of Streptomyces thermoviolaceus OPC-520,were cloned and sequenced. The chi35 and chi25 genes were arranged in tandem and encoded deduced proteins of 39,762 and 28,734 Da, respectively. Alignment of the deduced amino acid sequences demonstrated that Chi35 has an Nterminal domain and a catalytic domain and that Chi25 is an enzyme consisting of only a catalytic domain. Amino acid sequences of the catalytic domains of both enzymes, which are highly similar to each other, suggested that these enzymes belong to the family 19 chitinases. The cloned Chi35 and Chi25 were purified from E. coli and S. lividans as a host, respectively. The optimum pH of Chi35 and Chi25 were 5-6,and the optimum temperature of Chi35 and Chi25 were 60 and 70℃, respectively. Chi35 bound to chitin, Avicel, and xylan. On the other hand, Chi25 bound to these polysaccharides more weakly than did Chi35. These results indicate that the N-terminal domain of Chi35 functions as a polysaccharide-binding domain. Furthermore, Chi35 showed more efficient hydrolysis of insoluble chitin and stronger antifungal activity than Chi25. In the polysaccharidebinding domain of Chi35,there are three reiterated amino acid sequences starting from C-L-D and ending with W, and the repeats were similar to xylanase (STX-I) from the same strain. However, the repeats did not show sequence similarity to any of the known chitinbinding domains and cellulose-binding domains.
- 社団法人日本農芸化学会の論文
- 2000-11-23
著者
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WATANABE Takeshi
Department of Molecular Immunology, Medical Institute of Bioregulation, Kyushu University
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Inamori Yoshihiko
Department Of Microbiology Osaka University Of Pharmaceutical Sciences
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Hatano Naoya
Osaka University Of Pharmaceutical Sciences
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TSUJIBO Hiroshi
Osaka University of Pharmaceutical Sciences
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MIYAMOTO Katsushiro
Osaka University of Pharmaceutical Sciences
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INAMORI Yoshihiko
Osaka University of Pharmaceutical Sciences
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TSUJIBO Hiroshi
Department of Microbiology, Osaka University of Pharmaceutical Sciences
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OKAMOTO Takashi
Osaka University of Pharmaceutical Sciences
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MITSUTOMI Masaru
Biological Sciences, Faculty of Agriculture, Saga University
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Mitsutomi Masaru
Biological Sciences Faculty Of Agriculture Saga University
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Tsujibo H
Department Of Microbiology Osaka University Of Pharmaceutical Sciences
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Miyamoto K
Central Research Laboratories Osaka University Of Pharmaceutical Sciences
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Watanabe Takeshi
Department Of Applied Biological Chemistry Faculty Of Agriculture Niigata University
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Inamori Yoshihiko
Osaka College Of Pharmacy
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Watanabe Takeshi
Department Of Agricultural Chemistry Faculty Of Agriculture Tohoku University
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Miyamoto Katsushiro
Osaka Univ. Of Pharmaceutical Sciences
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