Partial purification and study of kinetic enzyme reductase in blood cell decomposition of mice with induced atherosclerosis and inhibitory effect of some isoflavones.

ABSTRACT The reseach includes isolation and study some kinetic characteristics of aldose reductase (AR2) from blood cells hemolysate for miceinduced atherosclerosis and normal mice. Two peaks of protein were isolated using ion exchang (DEAE Cellulose) chromatography . The specific activities of AR2 enzyme for these peaks were determined for normal and induced- atherosclerosis mice . The results showed 0.77 and 0.3 unit/mg protein for the two peaks of normal mice and 4.17 and 1.62 unit/mg protein for the two peaks induced- atherosclerosis mice. Peak I for AR2 appeared to be a high specific activity .The optimum conditions for peak I of the partial purified AR2 of blood cells hemolysate activity for induced- atherosclerosis mice were obtained and the results as follows : using (14 mmol /L) of glucose as a substrate , using phosphate sodiume buffer at (6.8 pH) , an optimum reaction time at (10) minutes at (25C) with volume of enzyme extract (200L ), a concentration of 10 mM of some positive ions which were indicat that manganes ions have the lowest inhibition effect for AR2 . Using LineWeaver-Burk plot , the values of maximum velocity (Vmax) and Michael ,s-Menton constant (Km) were 2U/ ml and 33.33mmol/ L respectively . Also inhibitory effect of some isoflavone compounds( Daidzein and Gensitein )were studied on the enzyme activity , and their results give different inhibitory ratios for this enzyme, the type of inhibition was obtained noncompetitive in hibition for these compounds.