Extraction and Purification and Activity Determination Conditions Optimization of α-Amylase Inhibitor in White Kidney Bean
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Abstract:
In order to develop a simple, effective and low-cost extraction and purification process of α-amylase inhibitor (α-AI) and explore the optimal conditions for its activity test, white kidney beans were used as raw materials to extract α-AI by water extraction in this study. After studying the conditions for the determination of α-AI activity by 3,5-dinitrosalicylic acid (DNS) micropore method, the optimal process conditions for α-AI extraction were determined by orthogonal test on the basis of single factor and the α-AI crude extract was purified by acid precipitation-alcohol precipitation, ultrafiltration and dialysis. The results indicated that the optimal conditions for the determination of α-AI activity were: determination wavelength of 482 nm, DNS dosage of 80 μL, reaction time of 15 min, and the rest time was 10 min. And the optimum process for α-AI extraction were: Extraction temperature of 45 ℃, extraction time of 2.5 h, extraction pH of 6.6 and the liquid-solid ratio was 7.5. Moreover, the α-AI with high purity and activity was obtained after purification, which with an IC50 of 0.89±0.04 mg/mL and a yield of 1.92±0.23%. The extraction and purification process may be can provide some guiding significance for the low-cost industrial production of α-AI.
