[Conditions for isolation of cholesterol oxidase from Actinomyces lavendulae mycelia].

The isolation conditions, composition and certain properties of cholesterol oxidase from the mycelium Actinomyces lavendulae were studied. Selective extraction of the enzyme occurred upon the mycelium extraction by the Tween-80 containing buffer. The specific activity of cholesterol oxidase thus obtained exceeded seven-fold that of the enzyme isolated via other methods. The optimal concentration of the detergent upon extraction was 0.15%. By disc-electrophoresis it was demonstrated that the preparation isolated from disrupted cells contained, in addition to cholesterol oxidase, 16 other protein components that had no enzymatic activity whereas the Tween-80 extracted preparation included no more than 7 fractions. The study of the relationship between activity of the resultant cholesterol oxidase and the substrate concentration and pH demonstrated that optimal conditions for the enzyme action were at pH 7.0-7.5 and cholesterol oxidase concentration of 0.0316 mM.