Abstract
An α-amylase with molecular weight of 73 kDa was purified from midgut of Dociostaurus maroccanus using ammonium sulfate precipitation, gel filtration, and ion exchange chromatography. The optimal pH and temperature were 6 and 45 °C, respectively. As calculated using Lineweaver–Burk plots, the Km was about 0.62 mM and the Vmax was 1.113 (μmol/min/mg protein). Mn2+, Hg+, Zn2+ and ethylene diamine tetraacetic acid (EDTA) decreased α-amylase activity of D. maroccanus, whereas the addition of K+, Na+, Fe2+, Ba+, Mg2+, Ca2+ and Co2+ increased enzyme activity. Alpha-amylase inhibitors (AI1, AI2) with molecular weights of 43 kDa and 29 kDa, respectively, were purified from the seeds of Phaseolus vulgaris and its inhibitory effect on purified α-amylase of D. maroccanus was investigated. These inhibitors inhibited the D. maroccanus gut α-amylase activity significantly.
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