Purification and determination the molecular weight and study the kinetic properties of Glucose-6-phosphate dehydrogenase from Diabetic patients

Abstract

This study was conducted to purifity Glucose 6-phosphate dehydrogenase from diabetic patients by gel filtration technique on Sephadex G100 and determine the molecular weight of enzyme, concentration and kinetic constant [Km, Vmax], study the effect of optimum temperature, substrate, pH. The study includes [60] patients with diabetes mellitus and [60] healthy individual as control. The activity of G6PD was measured after precipitated by [75%] Ammonium Sulfate concentration, using [Tris-HCl] buffer at pH 8.2. The specific activity was calculated [21.5UI\mg], total activity [706.8UI], number of purification [3.45] enzyme yield [23.188%] and enzyme activity [17.67UI\ml]. and the molecular weight were [57.82] k.D. The effect of substrate concentration was found to be increased with increasing substrate concentration. The optimum pH was found to be [8.4] and the optimum temperature was [38C]. Michaelis- Menten [Km] value for the enzyme was [3.33mM] and Vmax value was [0.263IU\ml ].