Simple Purification and Improvement of Stability of Alcohol Oxidase from a Methanol Yeast,Candida boidiniiS2, by Forming an Enzyme–Azide Complex

Abstract

It was found that azide bound to alcohol oxidase non-covalently and caused a color change from yellow to red. Alcohol oxidase was purified with a high yield from methanol-limited chemostat-grown cells of Candida boidinii S2 as an enzyme-azide complex by a simple procedure. That is, a cell-free extract was prepared from cells treated with a cationic detergent, Cation M2, and alcohol oxidase amounted to more than 80% of the soluble protein. Inactivation of the enzyme by H2O2 and aldehydes was decreased in the complex. The results were discussed as to the production of various aldehydes with the yeast cells.