Cloning of Pseudomonas fluorescens Carboxylesterase Gene and Characterization of Its Product Expressed in Escherichia coli
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概要
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A gene (estC) coding for an esterase (esterase III) of Pseudomonas fluorescens was cloned into Escherichia coli JM83. DNA sequencing showed a single open reading frame with GTG as a translation initiation codon for esterase III. This was confirmed by N-terminal amino acid sequence analysis of the purified esterase III protein from an E. coli clone. The promoter sequence and a potential Shine-Dalgarno sequence were followed by the coding sequence of the estC gene. The amino acid sequence deduced from the nucleotide sequence contains the consensus active site sequence, G-X-S-X-G, of serine esterase. The esterase III expressed in an E. coli clone was purified by ion-exchange chromatography and gel filtration. The native form of the enzyme was a monomer with a molecular weight of 41,000. The results of substrate specificity and the inhibitor studies suggest that this enzyme is a carboxylesterase (EC 3.1.1.1) and a serine residue is present at the active site of the enzyme.
- 社団法人日本農芸化学会の論文
- 1994-01-23
著者
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Park Seonyang
韓国
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PARK Seonyang
Department of Internal Medicine, Seoul National University Hospital
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YOO Ook
Department of Biological Science and Engineering, The Korea Advanced Institute of Science
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Park Sung
Department Of Laboratory Medicine Seoul National University College Of Medicine
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Kim Y
Ewha Womans Univ. Seoul Kor
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Choi Kang-duk
Animal Resources Research Center Kon-kuk University
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Yoo O
Department Of Life Science Korea Advanced Institute Of Science And Technology
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Yoo Ook
Department Of Biological Science
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Lee Hwanghee-blaise
Department Of Life Science Korea Advanced Institute Of Science And Technology
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Park S
Department Of Laboratory Medicine Seoul National University College Of Medicine
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Kim Youn-Sung
Department of Life Science, Korea Advanced Institute of Science and Technology
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Kim Youn-sung
Department Of Chemistry Seoul National University
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Park Seonyang
Department Of Laboratory Medicine Seoul National University College Of Medicine
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