Extraction and Purification and Characterization of Fungel Alpha-amylase Inhibitors From Local Maize Variety May 70

Ahmed Khalaf, Anwer; Zeki Naji, Ethar

	Extraction and Purification and Characterization of Fungel Alpha-amylase Inhibitors From Local Maize Variety May 70
Journal of Tikrit University For Agriculture Sciences
Article 1, Volume 16, Issue 2, 2016, Pages 209-225
Authors
Anwer Ahmed Khalaf; Ethar Zeki Naji
Abstract
Fungelα-amylase inhibitors were extracted from May 70 with 0.2 M sodium acetate buffer, pH 6 , then concentrated by using different saturation percentage of ammonium sulfate from extracted . The results showed that precipitation with ammonium sulphate 60% was more suitable, scoring inhibitor units and specific activity amounted to 10.62 units⁄ml, 12.45 units⁄ mg protein respectively .Then purified by ionexchange chromatography using DEAE-Cellulose, which identified peaks possessed effective in the inhibition of amylase A.niger in the recovery stage with purification fold of 5.1 , and recovery amounted 35.66% . purification has been completed on a gel filtration Sephadex G-100 column , The obtained purification fold was 8.25 , and recovery amounted 28.62%.Results showed that the characterization of amylase inhibitors molecular weight inhibitor purified reached at 29500 Kiloudalton appreciation manner gel filtration . The study showed that the incubating -amylase inhibitor purified with a 30-minute task to obtain a higher percentage of inhibition in the conditions used in this study .And gave determined highest effective inhibition toward the enzyme alpha - amylase at a temperature of 30 ˚ C where the effectiveness inhibitor 4.55 U / mL , while amylase with purified inhibitor at a temperature of 30 ˚ was effective inhibition 3.77 units / mL , with an incidence rise of the effectiveness of amylase when his incubating with purified inhibitor temperature 70 ˚ C , and when studying the degree of stability purified inhibitor after including temperature (90-30) ˚ C for 30 minutes was observed at the temperatures (50-30) ˚ C followed by a gradual decrease in efficiency with increased temperature , with retention activity for 80% of the inhibitor at degree 70 ˚C followed by decrease until it reached a degree 90 ˚ C to 60% . these the results can reflect higher thermal stability of these inhibitors . Optimum pH for linking of purified inhibitor with amylase is around 9-6 .The higher the stability of the purified inhibitor was at pH 7 , the purified inhibitor gave the highest effective inhibition and were more stable toward the basal numbers .
Keywords
niger Alpha; amylase inhibitors; Maize; niger amylase

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