Purification and properties of pyranose oxidase from basidiomycetous fungus No. 52.

Abstract

Pyranose oxidase was purified from the crude extract of basidiomycetous fungus No. 52 about 74-fold to a single protein band on polyacrylamide gel electrophoresis with an overall yield of 23%. The molecular weight of the native enzyme was about 300, 000 on gel filtration and 69, 000 on SDS-polyacrylamide gel electrophoresis. Its isoelectric point was pH 6.3. The enzyme activity showed high stability over a wide pH range of 5.5-9.0 and below 50°C, and 60% of its initial activity remained even after heating at 70°C. The enzyme was inactivated by Ag+, Hg2+, and Cu2+. The enzyme contained FAD covalently bound to the protein and was a glycoprotein containing about 0.7% carbohydrate.