Method for the purification of cholesterol oxidase

Abstract

A method for the purification and recovery of cholesterol oxidase is disclosed. Cholesterol oxidase is purified by a method including the steps of: (a) contacting a solution of the cholesterol oxidase to be purified with a chromatography composition comprising an unbound water-insoluble steroid which is a nonsubstrate for cholesterol oxidase so as to adsorb the cholesterol oxidase on the steroid; (b) separating the solution from the steroid; and (c) contacting the steroid with an eluting solution containing a surfactant at a concentration between 0.001 percent and 10 percent so as to recover the adsorbed cholesterol oxidase. In a preferred embodiment, the solution containing the cholesterol oxidase to be purified is a microorganism growth medium having cholesterol oxidase activity and, prior to the step of contacting the solution with a steroid, the solution is contacted with an ion-exchange chromatography material. In yet another embodiment, the eluting solution containing the surfactant also contains a metal salt of a concentration of at least about 0.005 molar. There is also disclosed a method for forming a packed bed of a composition comprising the unbound steroid which comprises the step of first suspending the steroid in a water-miscible organic solvent.

Dependencies are inferred from the claim text.