A Thermostable Histamine Oxidase from Arthrobacter crystallopoietes KAIT-B-007(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
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概要
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A thermostable histamine oxidase (EC 1.4.3.-) was found in cells of Arthrobacter crystallopoietes KAIT-B-007 isolated from soil. The enzyme was purified about 715-fold over the cell free extracts with a yield of 55% by ammonium sulfate fractionation and various column chromatographies. The purified enzyme was homogeneous on polyacrylamide gel-electrophoresis (native-PAGE). When the enzyme was kept at 65℃ and 70℃ for 10min, the activity was fully stable at 65℃ and decreased to 9% of the initial level at 70℃. The enzyme was very thermostable. The optimum pH for histamine oxidase activity was found to be at 9.0, and the enzyme was stable over the pH range of 6 to 9. The purified enzyme showed a single protein band on SDS-PAGE and its molecular mass was estimated to be about 81kDa. The enzyme showed potent activity toward histamine, whereas it was inactive toward putrescine, cadaverine, spermine, and spermidine. Histamine oxidase was inhibited by N,N-diethyldithiocarbamate (DDTC). The inactive enzyme was restored with Cu^<2+> to 65% of the initial activity, but Cu^+ did not enhance the enzyme activity. It is suggested that Cu^<2+> is essential for expression of histamine oxidase activity. The enzyme was a copper-containing protein having one atom of copper per mol of the enzyme protein as a result of atomic absorption analysis. The N-terminal amino acid sequence of the purified enzyme was different from that of histamine oxidase from Arthrobacter globiformis IFO12137.
- 社団法人日本生物工学会の論文
- 2004-02-25
著者
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MATSUMOTO Kunio
Department of Tumor Dynamics and Regulation, Cancer Research Institute, Kanazawa University
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YAMAMURA Akira
Department of Applied Chemistry, Faculty of Engineering, Kanagawa Institute of Technology
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SEKIGUCHI Yoshinori
Department of Applied Chemistry, Faculty of Engineering, Kanagawa Institute of Technology
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MAKITA Hiroko
Department of Biochemistry, Faculty of Engineering, Yokohama National University
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Matsumoto Kunio
Department Of Applied Chemistry Faculty Of Engineering Kanagawa Institute Of Technology
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Makita Hiroko
Department Of Chemical Engineering Science Graduate School Of Engineering Yokohama National Universi
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Yamamura A
Department Of Applied Chemistry Faculty Of Engineering Kanagawa Institute Of Technology
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Yamamura Akira
School Of Materials Science Japan Advanced Institute Of Science & Technology
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Yamamura Akira
Department Of Applied Chemistry Faculty Of Engineering Kanagawa Institute Of Technology
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Sekiguchi Yoshinori
Department Of Applied Chemistry Faculty Of Engineering Kanagawa Institute Of Technology
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