The reaction rate of glucose oxidase was determined at different glucose concentration ( 20 , 12.5 , 10 , 5 , 3.33 , 2.5 ) % of glucose under optimum conditions of pH , temperature and incubation time . the glucose oxidase velocity (enzyme activity per unit time) was determined at each substrate concentration and the values of Km and Vmax were determined by plotting Lineweaver–Burk plot