Cloning and Heterologous Expression of a β-Fructofuranosidase Gene from Arthrobacter globiformis IFO 3062, and Site-Directed Mutagenesis of the Essential Aspartic Acid and Glutamic Acid of the Active Site (ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHN
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概要
- 論文の詳細を見る
We have cloned the gene encoding a β-fructofuranosidase from Arthrobacter globiformis IFO 3062, and subsequently, the gene was heterologously expressed in Escherichia coli. This β-fructofuranosidase gene encodes a protein of 548 amino acid residues with a calculated molecular mass of 60,519 Da. We have examined the roles of three residues of A. globiformis IFO 3062 β-fructofuranosidase by site-directed mutagenesis, and found that aspartic acid 130 and glutamic acid 392, which are two of the apparent catalytic residues, are essential for hydrolase activity. This study provides the first experimental evidence showing that these two amino acid residues of β-fructofuranosidase play a critical role in hydrolyzing sucrose.
- 社団法人日本生物工学会の論文
- 2004-04-25
著者
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Ito Hiroyuki
Department Of Internal Medicine Division Of Cardiovascular And Respiratory Medicine Akita University
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Matsui Hirokazu
Department of Bioscience and Chemistry, Faculty of Agriculture, Hokkaido University
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森 春英
北海道大学大学院農学研究院応用生命科学部門
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WATANABE Kenji
Department of Kampo Medicine, Keio University, School of Medicine
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Watanabe K
Faculty Of Pharmaceutical Science School Of Medicine Fukuoka University
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Win T
Department Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Win Than
Department Of Quantum Science And Energy Engineering Graduate School Of Engineering Tohoku Universit
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Matsui H
Division Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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ISONO NAOTO
Department of Applied Bioscience, Graduate School of Agriculture, Hokkaido University
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TOCHIHARA TAKASHI
Department of Applied Bioscience, Graduate School of Agriculture, Hokkaido University
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KUSNADI YUYUS
Department of Applied Bioscience, Graduate School of Agriculture, Hokkaido University
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OBAE KAZUHIRO
Asahikasei Chemicals Corporation
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Ito Hiroyuki
Division Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Matsui Hirokazu
Dept. Biosci. Chem. Fac. Agric. Hokkaido Univ.
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Matsui Hirokazu
Department Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Matsui Hirokazu
Hokkaido Univ. Sapporo Jpn
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Watanabe Katsunori
Department Of Bioresources Chemistry Faculty Of Horticulture Chiba University
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Watanabe K
Tokyo Institute Of Technology Faculty Of Bioscience And Biotechnology Department Of Biological Scien
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Watanabe K
Division Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Isono N
Hokkaido Univ. Sapporo Jpn
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Kusnadi Yuyus
Department Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Tochihara T
Department Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Watanabe K
Research Institute For Food Science Kyoto University:(present Address)department Of Agricultural Che
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Watanabe K
Department Of Applied Biological Sciences Faculty Of Agriculture Saga University
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Watanabe K
Marine Biotechnol. Inst. Iwate Jpn
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Ito H
Division Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Ito Hiroyuki
Department Of Cardiology Akita University
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Isono Naoto
Department Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Watanabe K
Department Of Chemistry And Life Science College Of Bioresource Sciences Nihon University
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Tochihara Takashi
Department Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Matsui Hirokazu
Laboratory Of Biochemistry Research Faculty Of Agriculture Hokkaido University
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Watanabe Kenji
Department Of Gastroenterology Osaka City University Graduate School Of Medicine
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Watanabe Kenji
Department Of Gastroenterology Graduate School Of Medicine Osaka City University
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Watanabe Kenji
Department Of Anatomy Fukui Medical School
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Ito Hiroyuki
Department Of Anesthesiology Tokyo Medical And Dental University Graduate School Of Medical And Dent
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Watanabe Keiichi
The Graduate School Of Science And Technology Kobe University
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Win Than
Department Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Watanabe Kenji
Department Of Agricultural Chemistry College Of Agriculture And Veterinary Medicine Nihon University
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MATSUI Hirokazu
Department of Agricultural Chemistry, Faculty of Agriculture, Hokkaido University
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ISONO Naoto
Department of Sustainable Resource Science, Graduate School of Bioresources, Mie University
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