Preparation of Aldehyde Oxidase in Its Native and Deflavo Forms

Abstract

Abstract Aldehyde oxidase (EC 1.2.3.1) has been purified by a modification of a previously reported procedure and the FAD prosthetic group has been removed by treatment with calcium chloride and calcium acetate. The deflavo enzyme so obtained is devoid of N1-methylnicotinamide oxygen reductase activity but can be reconstituted by a short incubation with FAD. The different activities of native and deflavo enzymes using oxygen, cytochrome c, potassium ferricyanide, nitro blue tetrazolium, dichlorophenolindophenol, and FAD as electron acceptors and N1-methylnicotinamide as substrate are compared.