Cloning and Nucleotide Sequence of the β-D-Glucosidase Gene from Bifidobacterium breve clb, and Expression of β-D-Glucosidase Activity in Escherichia coli
スポンサーリンク
概要
- 論文の詳細を見る
Genomic DNA encoding a β-D-glucosidase (EC 3.2.1.21), which has β-D-fucosidase activity, was cloned from Bifidobacterium breve clb. We sequenced a 1.9-kbp cloned DNA fragment that contained a single open reading frame encoding 460 amino acids with a calculated molecular mass of 51,513 Da. A putative ribosome binding site was found 5 bp upstream of the initiation codon. The amino acid sequence of this β-D-glucosidase from Bifidobacterium breve clb had 46% identity with that of β-glucosidase from Microbispore bispore. The enzyme of Bifidobacterium breve clb was expressed in Escherichia coli. A cell-free extract prepared from the recombinant strain showed 80 to 90-fold more β-D-glucosidase activity than that from Bifidobacterium breve clb. The recombinant enzyme was purified to homogeneity from cell-free extracts of the recombinant strain using 4 column chromatographies. The recovery of enzyme from the recombinant strain was about 138-fold-higher than that of Bifidobacterium breve clb. The enzymatic properties were similar to those of Bifidobacterium breve clb. For application of this recombinant enzyme, we attempted to synthesize a disaccharide that seemed to be specifically assimilated by Bifidobacteria using the condensation activity of the enzyme.
- 社団法人日本農芸化学会の論文
- 1996-12-23
著者
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YAMAMOTO Kenji
Department of Medical Ecology and Informatics, Research Institute, International Medical Center of J
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YANO Toshihiro
Department of Material Science, Graduate School of Science, Osaka City University
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TANAKA Keiko
Department of Chemistry, Faculty of Science, Nara Women's University
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Kumagai Hidehiko
Research Institute For Bioresources And Biotechnology Ishikawa Prefectural University
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TAMAKI HISANORI
Department of Food Science and Technology, Kyoto University
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KUMAGAI HIDEHIKO
Department of Food Science and Technology, Kyoto University
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NUNOURA Naoki
Department of Food Science and Technology, Faculty of Agriculture, Kyoto University
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OHDAN Kohji
Department of Food Science and Technology, Faculty of Agriculture, Kyoto University
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INUI Masayuki
Research Institute of Innovative Technology for the Earth (RITE)
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YUKAWA Hideaki
Research Institute of Innovative Technology for the Earth (RITE)
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Kumagai Hidehiko
Department Of Food Science And Technology Faculty Of Agriculture Kyoto University
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Kumagai Hidehiko
Faculty Of Agriculture Kyoto University
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Tamaki H
Kyoto Univ. Kyoto Jpn
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Tamaki Hisanori
Division Of Integrated Life Sciences Graduate School Of Biostudies Kyoto University
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Tamaki Hisanori
Department Of Biochemical Science And Technology Faculty Of Agriculture Kagoshima University
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Yokoyama Tatsuo
Institute For Fermentation Osaka
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Kumagai Hidehiko
Res. Inst. For Bioresources And Biotechnology Ishikawa Prefectural Univ.
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Kumagai Hidehiko
Division Of Integrated Life Sciences Graduate School Of Biostudies Kyoto University
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Yano T
Department Of Food Science Ishikawa Agricultural College
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Ohdan K
Department Of Food Science And Technology Faculty Of Agriculture Kyoto University
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Ohdan Kohji
Department Of Food Science And Technology Facylty Of Agriculture Kyoto University
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Inui Masayuki
Research Institute Of Innovative Technology For The Earth
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Nunoura N
Department Of Food Science And Technology Faculty Of Agriculture Kyoto University:(present Address)r
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Nunoura Naoki
Department Of Food Science And Technology Faculty Of Agriculture Kyoto University Kitashirakawa
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Yukawa Hideaki
Research Institute Of Innovative Technology For The Earth
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Yano Toshihiro
Department Of Food Science Ishikawa Agricultural College
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Kumagai H
Department Of Agricultural Chemistry The University Of Tokyo
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Kumagai Hidehiko
Graduate School Of Biostudies Kyoto University
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Yano T
Research Laboratory Of Higashimaru Shoyu Co. Ltd.
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Yamamoto Kenji
Department Of Cardiovascular Surgery Kanagawa Cardiovascular And Respiratory Center
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Tanaka Keiko
Department Of Food Science And Technology Faculty Of Agriculture Kyoto University
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Tanaka Keiko
Department Of Anesthesiology Akashi Municipal Hospital
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Tamaki Hisanori
Industrial Research Institute Of Ishikawa
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KUMAGAI Hidehiko
Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University
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YANO Toshihiro
Department of Food Science and Technology, Kyoto University
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YANO Toshihiro
Department of Applied Chemistry, Nagoya Institute of Technology
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