Gene cloning and characterization of arylamine N-acetyltransferase from Bacillus cereus strain 10-L-2(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
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概要
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Bacillus cereus strain 10-L-2 synthesizes two arylamine N-acetyltransferases (Nat-a and Nat-b) with broad substrate specificities toward aniline and its derivatives. In southern blot analysis using probes encoding the NH_2-terminus of Nat-b and a conserved region of N-acetyltransferases, digested total DNA of strain 10-L-2 showed one positive band. We cloned and sequenced the gene encoding Nat-b. The NH_2-terminal amino acid sequence predicted from the open reading frame (768 base pairs) corresponded to that of purified Nat-b. The cloned Nat-b gene was expressed in Escherichia coli. The expressed enzyme (BcNAT) from the recombinant strain was partially purified and characterized. Nat-b from strain 10-L-2 and BcNAT from the recombinant strain were slightly different from each others in substrate specificity and thermo-stability. We examined the biotransformations of 2-aminophenols and phenylenediamines by the whole cells of the recombinant strain. The cells converted these compounds into their corresponding acetanilides. Only one amino group of phenylenediamines was acetylated. The cells utilized 4-nitroacetanilide as an acetyl donor instead of acetyl-CoA. 4-Aminoacetanilide was produced and 4-nitroaniline was released almost stoichiometrically.
- 社団法人日本生物工学会の論文
著者
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Aoki K
Department Of Applied Biological Chemistry Graduate School Of Agricultural Science Kobe University
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Takenaka Shinji
Department of Agrobioscience, Graduate School of Agricultural Science, Kobe University
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MURAKAMI Shuichiro
Department of Bioscience, Biotechnology, and Agrochemistry, Faculty of Agriculture, Meiji University
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AOKI Kenji
Department of Nutritional Management, Faculty of Nutritional Science, Sagami Women’s University
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Murakami S
Kikkoman Corp. Chiba Pref. Jpn
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Cheng Minyi
Department of Applied Biological Chemistry, Graduate School of Agricultural Science, Kobe University
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Cheng Minyi
Division of Biosystems Chemistry, Graduate School of Science and Technology, Kobe University
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Koshiya Atsushi
Department of Applied Biological Chemistry, Graduate School of Agricultural Science, Kobe University
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Cheng Minyi
Division Of Biosystems Chemistry Graduate School Of Science And Technology Kobe University
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Koshiya Atsushi
Department Of Applied Biological Chemistry Graduate School Of Agricultural Science Kobe University
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Murakami Shuichiro
Dep. Of Bioscience Biotechnology And Agrochemistry Fac. Of Agriculture Meiji Univ.
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Aoki Kenji
Department Of Nutritional Management Faculty Of Nutritional Science Sagami Women's University
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Takenaka Shinji
Department Of Agrobioscience Graduate School Of Agricultural Science Kobe University
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Aoki K
Department Of Agricultural Chemistry Faculty Of Agriculture Kobe University
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Murakami Shuichiro
Department Of Bioscience Biotechnology And Agrochemistry Faculty Of Agriculture Meiji University
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Aoki Kenji
Department Of Agricultural Chemistry Faculty Of Agriculture Kobe University
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MURAKAMI Shuichiro
Department of Biofunctional Chemistry, Faculty of Agriculture, Kobe University
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