Turbidimetric method for determination of glycogen phosphorylase activity and its use for estimation of equilibrium position of enzymic reaction.
A turbidimetric method has been developed for the continuous monitoring of the enzyme reaction catalyzed by glycogen phosphorylase. This method is based on the registration of the turbidity of glycogen solution at wavelengths above 300 nm. It has been shown that the increase in the turbidity is strictly proportional to the quantity of glucose 1-phosphate formed during the enzyme reaction. The method has the advantage of continuity, and it is suitable for determining the initial rate of catalytic synthesis or degradation of glycogen in a relatively simple and fast way. The kinetic experiments may be carried out under various conditions. The method of calculation of the overall equilibrium constant of the enzyme reaction catalyzed by glycogen phosphorylase has been elaborated. This method is based on the analysis of the dependence of the initial rate of the enzymic reaction on the proportion of the substrate of the forward reaction: [Pi]/( [Pi] + [G-1-P] ).
D000818:Animals; D007700:Kinetics; D009391:Nephelometry and Turbidimetry / Q000379:methods; D010763:Phosphorylase b / Q000032:analysis; D006005:Phosphorylases / Q000032:analysis*; D011817:Rabbits
D010763:Phosphorylase b; D006005:Phosphorylases