Carboxypeptidase Taq, a Thermostable Zinc Enzyme, from Thermus aquaticus YT-1 : Molecular Cloning, Sequencing, and Expression of the Encoding Gene in Escherichia coli
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概要
- 論文の詳細を見る
The gene for carboxypeptidase Taq, a thermostable metallo-carboxypeptidase from Thermus aquaticus YT-1, was cloned and sequenced. The gene comprised an open reading frame of 1, 536 base pairs with a GTG initiation codon and a TGA termination codon, which encodes a protein of 56, 210 Da consisting of 511 amino acid residues. The GTG initiation codon of the gene was replaced with ATG by site-directed mutagenesis, and then the gene was expressed in Escherichia coli. The enzyme purified from E. coli cells showed the same properties as those of carboxypeptidase Taq prepared from T. aquaticus cells. Analysis for metal ions bound to the enzyme found that one molecule of the enzyme contains one tightly bound zinc ion. Comparison of the entire sequence showed that the enzyme has no obvious sequence similarity to any other metallo-peptidases. However, a His-Glu-X-X-His sequence, which is a conserved sequence in the ac-tive site of zinc-dependent endopeptidases and aminopeptidases, was found at positions 276 to 280 of the enzyme. These findings suggest that carboxypeptidase Taq is a novel type of zinc-dependent metallo-carboxypeptidase.
- 社団法人日本農芸化学会の論文
- 1994-08-23
著者
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Ohta T
Hokkaido Food Processing Res. Center Hokkaido Jpn
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Lee S‐h
Columbia Univ. Ny Usa
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Mori Hiroaki
Department Of Agricultural Chemistry The University Of Tokyo
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Yoshimura Etsuro
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The U
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KAMINOGAWA Shuichi
Department of Food Science and Technology, College of Bioresource Sciences, Nihon University
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MATSUZAWA Hiroshi
Department of Biotechnology, The University of Tokyo
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Lee Sang-hyeon
Department Of Agricultural Chemistry The University Of Tokyo
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OHTA Takahisa
Department of Agricultural Chemistry, The University of Tokyo
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Kaminogawa Shuichi
Department Of Applied Biological Chemistry Division Of Agriculture And Agricultural Life Sciences Th
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Taguchi Hayao
Department of Agricultural Chemistry, The University of Tokyo
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Minagawa Etsuo
Department of Agricultural Chemistry, The University of Tokyo
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Taguchi H
Department Of Bioresource Development Hiroshima Prefectural University
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Ohta Takahisa
Department Of Agricultural Chemistry The University Of Tokyo
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Kaminogawa S
Dep. Of Food Sci. And Technol. Nihon Univ.
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MINAGAWA ETSUO
Research Center, Yotsuba Milk Products Co.Ltd.
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Minagawa Etsuo
Research Center Yotsuba Milk Products Co.ltd.
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Matsuzawa H
Department Of Bioscience And Biotechnology Aomori University
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Kaminogawa Shuichi
Department Of Agricultural Chemistry Faculty Of Engineering The University Of Tokyo
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Yoshimura Etsuro
Department Of Applied Biological Chemistry Graduate School Of Agricultural And Life Sciences The Uni
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Matsuzawa Hiroshi
Department Of Biotechnology And Biotechnology Research Center The University Of Tokyo
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Yoshimura Etsuro
Department of Agricultural Chemistry, The University of Tokyo
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Matsuzawa Hiroshi
Department of Bioscience and Biotechnology, Aomori University
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MATSUZAWA Hiroshi
Department of Agricultural Chemistry, The University of Tokyo
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