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Purification and Characterization of the Enzyme Cholesterol Oxidase from a New Isolate of Streptomyces sp.

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Abstract

An extracellular cholesterol oxidase (cho) enzyme was isolated from the Streptomyces parvus, a new source and purified 18-fold by ion exchange and gel filtration chromatography. Specific activity of the purified enzyme was found to be 20 U/mg with a 55 kDa molecular mass. The enzyme was stable at pH 7.2 and 50 °C. The enzyme activity was inhibited in the presence of Pb2+, Ag2+, Hg2+, and Zn2+ and enhanced in the presence of Mn2+. The enzyme activity was inhibited by the thiol-reducing reagents (DTT, β-mercaptoethanol), suggesting that disulfide linkage is essential for the enzyme activity. The enzyme activity was found to be maximum in the presence of Triton X-100 and X-114 detergents whereas sodium dodecyl sulfate fully inactivated the enzyme. The enzyme showed moderate stability towards all organic solvents except acetone, benzene, chloroform and the activity increased in the presence of isopropanol and ethanol. The K m value for the oxidation of cholesterol by this enzyme was 0.02 mM.

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Acknowledgments

This study was financially supported by the Council of Scientific and Industrial Research, India.

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Authors and Affiliations

  1. Fermentation Technology Division, Central Drug Research Institute (CSIR), Chattar Manzil, P.O. Box 173, Lucknow, 226 001, Uttar Pradesh, India

    Vandana Praveen, Akanksha Srivastava & C. K. M. Tripathi

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  1. Vandana Praveen
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  2. Akanksha Srivastava
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  3. C. K. M. Tripathi
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Correspondence to C. K. M. Tripathi.

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Praveen, V., Srivastava, A. & Tripathi, C.K.M. Purification and Characterization of the Enzyme Cholesterol Oxidase from a New Isolate of Streptomyces sp.. Appl Biochem Biotechnol 165, 1414–1426 (2011). https://doi.org/10.1007/s12010-011-9360-6

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