Purification and Characterization of Extra-Cellular Cholesterol Oxidase From Rhodococcus sp. PTCC 1633

In this study to isolate cholesterol oxidase (Cho) producing microorganism, a bacterium with high ability to produce Cho was found. It was identified with morphological, biochemical and molecular methods as a new species of Rhodococcus. Rhodococcus sp. PTCC 1633 has both extra-cellular and intracell...

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Published in:Biotechnology (Faisalābād, Pakistan) Pakistan), 2008-12, Vol.7 (4), p.751-756
Main Authors: Yazdi, M.T., Yazdi, Z.T., Ghasemian, A., Zarrini, G., Olyaee, N.H., Sepehrizad, Z.
Format: Article
Language:English
Subjects:
Rhodococcus
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Summary:In this study to isolate cholesterol oxidase (Cho) producing microorganism, a bacterium with high ability to produce Cho was found. It was identified with morphological, biochemical and molecular methods as a new species of Rhodococcus. Rhodococcus sp. PTCC 1633 has both extra-cellular and intracellular Cho. It was cultured in optimized condition and extra-cellular Cho concentrated by Three Phase Partitioning (TPP) with a recovery of 85%. The concentrated enzyme was purified by one step ion exchange chromatography. On SDS-PAGE the purified enzyme showed a molecular weight of about 55 kDa. The enzyme was active in a wide rang of pH and temperature with an optimum pH and temperature of 7.0-7.5 and 40 degree C, respectively, for activity. The K sub(m) value for this enzyme was 15 mu M. A thermal stability experiment showed high stability at 40 degree C in 24 h.
ISSN:1682-296X
DOI:10.3923/biotech.2008.751.756