Identification of Novel Cholesterol Oxidase from Rhodococcus Erythropolis ATCC 4277

Cholesterol oxidase enzyme is important in industry. The nocardioform actinomycetes and rhodococcus species have been used for cholesterol oxidase production. A novel cholesterol oxidase enzyme was investigated in Rhodococcus erythropolis sp strain using biochemical and molecular techniques. Cell production was increased rapidly, using M65 medium and media analyses demonstrated that the maximal growth rate occurs at the primary 36 hours. However, the enzyme production starts after a log phase and continues at the stationary phase in maximal rate after 60 hours of cultivation. A lower level of enzyme activity was detected by this strain compared to the other strains with commercial applications. The prepared Immune antibody from standard cholesterol oxidase demonstrated no cross reaction against the cholesterol oxidase of this strain using western blot examination. However a 240bp fragment was observed in PCR amplification with a couple of primers designed with a partial sequence of a novel cholesterol oxidase. The sequences of this fragment demonstrated 99% alignment, compared to a novel cholesterol oxidase from Nocardioform Microorganism using Megaalign technique in DNA star software.