Purification and immobilization of sulfhydryl oxidase

Abstract

Sulfhydryl oxidase is isolated in substantially purified form by precipitating from whey a crude impure fraction of sulfhydryl oxidase, dissolving the crude sulfhydryl oxidase in a dilute neutral buffer solution, equilibrating to allow for dissociation of the dissolved sulfhydryl oxidase, separating the equilibrated sulfhydryl oxidase from molecularly larger materials, concentrating the resultant separated sulfhydryl oxidase and separating the concentrated sulfhydryl oxidase from molecularly smaller materials to isolate a substantially purified sulfhydryl oxidase enzyme having a specific activity at least about fifty times greater than the crude enzyme and about 3000 times greater than that in skim milk. The enzyme may be immobilized, for example, by attaching it to an inert support. Activity of the immobilized enzyme may be regenerated by treatment with an aqueous solution of ferrous ions.