Purification and Characterization of Extracellular 3β-Hydroxysteroid Oxidase produced by Streptoverticillium cholesterolicum

摘要: 3β-Hydroxysteroid oxidase activity was detected in the broth filtrate of strain H 1109 MY 12. Based on taxonomic studies, this shown to be a new species genus Streptoverticillium and name Stv. cholesterolicum is proposed for strain. The enzyme purified by ammonium sulfate precipitation affinity column chromatography crystalline cholesterol, product showed single band SDS-polyacrylamide gel electrophoresis. optimum at pH 7.0-7.5 stable over rather wide range 4.0 12.5. Cholesterol dihydrocholesterol were oxidized rapidly. Km value oxidation cholesterol about 0.4mM. greately inhibited HgCl2, AgNO3. FeCl3 CuSO4 also inhibitory though lesser extents. Iodine N-bromosuccinimide completely concentrations less than 0.01 mM. Neither metal-binding agents nor p-chloromercuribenzoic acid had any effect enzyme. molecular weight estimated 56000 proved flavoprotein containing flavin adenine dinucleotide as prosthetic group.