تنقية و توصيف مثبطات ألفا-أميليز لعاب الإنسان من محصول الشعير Hordeum vulgare

Amylase inhibitors were purified by many sequential steps included concentration by gradual addition of ammonium sulfate at saturation ratios. ranged from 0 to 90% . The best ratio of saturation was found to be 70% as the specific activity and inhibition activity toward Human alpha-amylase(HAS) were the highest (8 U/mg and 6 U/ml respectively as compared to those of the rest ratios, the ratio of saturation with ammonium sulfate 60 % and then 50%, (5.8, 5.5)U/ml and(7‘ 7.7)U/mg respectively for inhibition activity and specific activity and for 40% ,30%20% saturation the inhibition activity and specific activity were(4‘ 4.8‘ 5) u/ml (5.8‘ 6‘ 6.6) u/mg respectively .The precepitation step was followed by ionic exchange chromatography technique by DEAE-cellulose column) 11x3)cm and the results showed that there was one peak with inhibition activity toward (HAS). Further purification steps were conducted using gel filtration on Sephacryl S-200 column (1.5 x 60 )cm; the purification folds was5.59 times with outcome of 46.5%.The results of alpha-amylase inhibitors characterization showed that the molecular weight was about 23.44 and 22.9 kDa as determined by electrophoresis and gel filteration respectively.