微孔板法筛选α-葡萄糖苷酶抑制剂的条件优化

Optimization of a microplate bioassay method of filtering alpha-glucosidase inhibitors

研究以微孔板检测法为基础,对α-葡萄糖苷酶抑制剂的抑制能力进行检测,以达到有效筛选适宜浓度的α-葡萄糖苷酶抑制剂用于制备降血糖类功能性食品的目的。试验针对酶浓度、检测波长、蔗糖底物浓度及酶反应时间进行反应条件优化。结果表明：试验中所用批次酶的最适浓度范围为2.0-3.0 mg/mL;在405,450,492,520,630nm 5种波长条件下,520nm和492nm波长检测信号最强;蔗糖底物浓度的合适范围应为20-40mmol/L;适宜的酶反应时间范围为20-30min。

The half inhibitory concentration （IC50） of alpha glueosidase inhibitors was detected based on a microplate assay method in this study in order to find the effective inhibitors for preparing hypoglycemic functional food. The reaction conditions were optimized by the four aspects, i.e., the enzyme concentration, the detection wavelength, the substrate concentration and the reaction time. The results showed that the optimum enzyme concentration ranged from 2.0 mg/mL to 3.0 mg/mL, and waves 520 nm and 492 nm were optimum among the five waves, 405, 450, 492, 520, 630 nm. Moreover, the suitable substrate concentration of sucrose ranged from 20 mmol/L to 40 mmol/L, and the appropriate reaction time was about 20-30 min.