Analysis of Catalytic Residues of Thermoactinomyces vulgaris R-47 α-Amylase II (TVA II) by Site-directed Mutagenesis
スポンサーリンク
概要
- 論文の詳細を見る
To confirm that the catalytic residues (Asp325,Glu354,and Asp421) are necessary for the hydrolysis of starch, pullulan, and cyclodextrins, we constructed TVA II mutated by site-directed mutagenesis. The mutated enzymes (D325N, E354Q, and D421N) had markedly reduced levels of activity, less than 0.006% of the wild type, indicating that these three residues are the catalytic sites for these substrates. Even E354D had reduced levels of activity, less than 0.05% of wild type. These four mutated enzymes retained a trace of activity. From the result of hydrolysis patterns for maltohexaose, in particular, D421N, unlike D325N and E354Q, catalyzed transglycosylation rather than hydrolysis. The results suggest that Asp421 could function to capture water molecules.
- 2000-12-23
著者
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TONOZUKA Takashi
Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture an
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殿塚 隆史
東京農工大学農学部
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Yokota T
Faculty Of Agriculture Shizuoka University
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YOKOTA Takehiro
Department of Applied Biological Science, Tokyo University of Agriculture Technology
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SHIMURA Yoichiro
Department of Applied Biological Science, Tokyo University of Agriculture Technology
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ICHIKAWA Kazuhiro
Department of Applied Biological Science, Tokyo University of Agriculture Technology
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SAKANO Yoshiyuki
Department of Applied Biological Science, Tokyo University of Agriculture Technology
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Yuuki T
Asahi Breweries Ltd. Ibaraki Jpn
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Tonozuka Takashi
Department Of Applied Biological Science Faculty Of Agriculture Tokyo University Of Agriculture And
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Ichikawa Kimihisa
Department Of Applied Microbiology Nippon Roche Research Center
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Shimura Y
Kitasato Univ. School Of Medicine Kanagawa Jpn
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Sakano Y
Department Of Applied Biological Science Faculty Of Agriculture Tokyo University Of Agriculture And
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Sakano Yoshiyuki
Department Of Applied Biological Science Faculty Of Agriculture Tokyo University Of Agriculture And
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Yokota Takao
Iss Science Project Office Institute Of Space And Astronautical Science Japan Aerospace Exploration
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Tonozuka Takashi
Forage Crop Breeding Unit National Agricultural Research Center For Kyushu Okinawa Region
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Ichikawa Kazuhiro
Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology
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SAKANO Yoshiyuki
Department of Agricultural Chemistry, Faculty of Agriculture, Tokyo Noko University
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