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Purification and Characterization of Cholesterol Oxidase Produced by Streptomyces polychromogenes IFO 13072.  

김현수 (계명대학교 자연과학대학 미생물학과)
성림식 (계명대학교 자연과학대학 미생물학과)
이경화 (계명대학교 자연과학대학 미생물학과)
이용직 (계명대학교 자연과학대학 미생물학과)
이인선 (계명대학교 자연과학대학 식품가공학과)
유대식 (계명대학교 자연과학대학 미생물학과)
Publication Information
Microbiology and Biotechnology Letters / v.30, no.2, 2002 , pp. 142-150 More about this Journal
Abstract
Streptomyces polychromogenes IFO 13072 was used as a strain producing cholesterol oxidase(EC 1.1.3.6). The conditions of cholesterol oxidase production were investigated. The optimum composition of medium for production of the enzyme was 1% dextrin, 0.5% casamino acid, 0.1% $KH_2$PO$_4$, 0.5% $NaNO_3$ and 0.05% $MgSO_4$(pH 7.3). The enzyme was purified specifically by cholesterol affinity column chromatography with a yield of 23.2%. The purified enzyme showed a single polypeptide on SDS-PAGE and the molecular weight was estimated about 52,000 daltons. The optimum pH and temperature of the cholesterol oxidase were pH 7.0 and $37^{\circ}C$, respectively. The enzyme was stable in the range of pH 6.0~7.0 and $25^{\circ}C$. The cholesterol oxidase activity was strongly inhibited by metal ions such as $Hg^{2+}$ and $Fe^{2+}$ and inhibitors such as dithiothreitol, mercaptoethanol and isonicotinic acid. The Michaelis constant(Km) for the cholesterol was found to be 25 mM by Lineweaver-Burk plot analysis.
Keywords
Cholesterol oxidase; Streptomyces polychromogenes; producing condition; purification; characterization; cholesterol affinity column chromatography;
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Times Cited By KSCI : 6  (Citation Analysis)
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