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

الملخص EN

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.