Cholesterol oxidase uses FAD to catalyze the oxidation of 3Beta-hydroxy steroids, which the enzyme then isomerizes into 3-ketosteroids. It is very similar to glucose oxidase in structure and function. It is used by microorganisms that are capable of using cholesterol as their sole source of energy. This particular structure is from the bacterium Streptomyces. The overall reaction is:

Cholesterol + O2 --------> Cholest-4-en-3-one + H2O2

Cholesterol oxidase shows great potential as an insecticide, because insect larval gut epithelial cells are rich in 3Beta-hydroxy sterols. Oxidation of these sterols weakens cell membranes and the insect dies. Bacterial cholesterol oxidase is also used as a diagnostic for detecting serum cholesterol levels.

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