Purification and Characterization of the Glucose Oxidase from Penicillium notatum

Abstract

This study aims to purification and characterization of the glucose oxidase enzyme from Penicillium notatum, the enzyme was purified by ammonium sulfate precipitation (60%), dialysis and gel filtration chromatography using sephadex G-200, A trial for the purification of glucose oxidase using gel filtration technique resulted in one type of glucose oxidase with specific activity of (62.382 U/mg) with (7.385 folds) purification. the purified glucose oxidase had a maximum activity at pH = 5.5, 45 °C, glucose oxidase was stable with pH values ranging between (5 – 6) and the enzyme was maintained the activity when it incubated into (25 -35) °C for 15 minutes, analyses of the glucose oxidase for molecular weight was carried out by PAGE and SDS-PAGE electrophoresis, which revealed 78 KDa, also molecular weight of the glucose oxidase was achieved by gel filtration technique and was found 87 KDa this means that enzyme consisting of only one subunit, the Km and Vmax value of glucoamylase (B) were (19.6 mM, 7.5 mM/min ) respectively using different concentration of glucose.