Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education of China), College of Chemistry & Chemical Engineering of Guangxi Normal University, Guilin 541004, PR China.
Food Chem. 2013 Dec 1;141(3):1854-9. doi: 10.1016/j.foodchem.2013.04.100. Epub 2013 May 9.
A novel strategy for screening α-glucosidase inhibitors (AGIs) from natural products by capillary electrophoresis (CE) with an immobilised enzyme microreactor was developed. In this approach, gold nanoparticles (AuNPs) was first covalently attached to surface of the pores of the porous polymer capillary monolith via the formation of an Au-S bond, and α-glucosidase was then simply and stably immobilised onto AuNPs through the strong affinity of gold for amino groups of the enzyme. In order to profiling the activity of the immobilised α-glucosidase, the natural substrate was hydrolyzed by it and the yield of product was determined by CE. The amount of covalently attached α-glucosidase to the monolith was calculated to be about 30.0 μg/mg. The immobilised enzyme exhibited 80% activity after 25 runs, and only lost 7.6% of activity after 6 runs within 31 days. Screening of AGIs present in extracts of natural products by the proposed method was demonstrated.
一种通过毛细管电泳（CE）用固定化酶微反应器从天然产物中筛选α-葡萄糖苷酶抑制剂（AGIs）的新策略被开发出来。在这种方法中，首先通过形成 Au-S 键将金纳米粒子（AuNPs）共价连接到多孔聚合物毛细管整体柱的孔表面上，然后通过金对酶的氨基的强亲和力，将α-葡萄糖苷酶简单且稳定地固定到 AuNPs 上。为了分析固定化α-葡萄糖苷酶的活性，用它水解天然底物，并通过 CE 测定产物的产量。计算出固定在整体柱上的α-葡萄糖苷酶的量约为 30.0 μg/mg。固定化酶在 25 次运行后表现出 80%的活性，并且在 31 天内的 6 次运行中仅损失了 7.6%的活性。通过所提出的方法证明了从天然产物提取物中筛选 AGIs 的存在。
