Purification and properties of the membrane-bound NADH oxidase of a facultatively anaerobic alkaliphile |
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| Authors: | Tomoko Hamada Takayoshi Wakagi Hirotaka Shiba N. Koyama |
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| Affiliation: | (1) Department of Chemistry, Faculty of Science, Chiba University, Yayoi, Inage-ku, Chiba 263-8522, Japan e-mail: nkoyama@scichem.c.chiba-u.ac.jp, JP;(2) Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan, JP;(3) Technical Development Department, Suntory Limited, Technological Development Center, Shimamoto, Mishima-gun, Osaka 618-0001, Japan, JP |
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| Abstract: | A membrane-bound NADH oxidase of an anaerobic alkaliphile, M-12 (a strain of Amphibacillus sp.), was solubilized with decanoyl N-methylglucamide and purified by chromatography on DEAE-Sepharose and hydroxyapatite. The purified enzyme appears to consist of a single polypeptide component with an apparent molecular mass of 56 kDa. The enzyme catalyzed the oxidation of NADH with the formation of H2O2 and exhibited a specific activity of 46 μmol NADH min–1 (mg protein)–1. NADPH did not serve as a substrate for the enzyme. The K m for NADH was estimated to be 0.05 mM. The enzyme exhibited a pH dependence for activity, with a pH optimum at approximately 9.5. The enzyme required a high concentration of salt and exhibited maximum activity in the presence of 600 mM NaCl. Received: 3 August 1998 / Accepted: 23 December 1998 |
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| Keywords: | NADH oxidase Membrane Alkaliphile Halophile |
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