Reduction of 2-carboxy-4'-dimethylaminoazobenzene by Bacillus diaphorase.
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概要
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2-Carboxy-4′-dimethylaminoazobenzene (CDMAB) was reductively cleaved into its two component primary arylamines by <I>Bacillus</I> diaphorase together with NADH (pH 6.1, 25 °C). The reductive cleavage was found to proceed stoichiometrically by a spectrofluorometric assay of fluorescent <I>o</I>-aminobenzoic acid and a fluorescent product formed between <I>N</I>,<I>N</I>-dimethyl-<I>p</I>-phenylenediamine and fluorescamine. Parallel lines were obtained in Lineweaver–Burk plots of initial rate vs. CDMAB concentration for the reduction. This experimentally observed kinetic behavior is consistent with a reaction scheme in favor of the Ping Pong mechanism in which CDMAB is reduced by a reduced form of diaphorase. From a kinetic formula, the apparent <I>K</I><SUB>m</SUB>(CDMAB), <I>K</I><SUB>m</SUB>(NADH), and <I>V</I><SUB>max</SUB> values are 2.14×10<SUP>−5</SUP> mol dm<SUP>−3</SUP>, 7.35×10<SUP>−5</SUP> mol dm<SUP>−3</SUP>, and 1.8×10<SUP>−7</SUP> mol dm<SUP>−3</SUP> min<SUP>−1</SUP> per mg of protein, respectively.
- 公益社団法人 日本化学会の論文
著者
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Hayashi Hideki
Department Clinical Pharmacology & Genetics, School of Pharmaceutical Sciences, University of Shizuoka
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Yatome Chizuko
Department of Chemistry, Faculty of Engineering, Gifu University
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Ogawa Toshihiko
Department of Chemistry, Faculty of Engineering, Gifu University
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