In this work, α-glucosidase was immobilized on a type of nanoscaled magnetic materials Fe₃O₄@ZIF-67 to construct a biomicroreactor for rapid screening α-glucosidase inhibitors. The parameters that affected the immobilization efficiency were investigated. Under optimal conditions, the amount of α-glucosidase loaded was up to 79.07 μg·mg^-1. Then, the enzyme biomicroreactor was immobilized by an external magnetic field in a tube connecting HPLC and micro-injection pump at both ends to form a magnetic online screening system. The screening flow rate and eluent organic solvent ratio during the screening process were optimized. XinYang MaoJian crude tea extract was tested to verify this online screening method, and three inhibitors (catechin, epigallocatechin gallate and epicatechin gallate) were fished by this magnetic online screening system. Based on the classical pNPG method, the inhibitory activities on α-glucosidase were further verified and studied. Compared with the traditional ligand fishing methods, the screening method established in this work can integrate screening, elution and analysis. It can simply, efficiently and directly screen and identify potential α-glucosidase inhibitors from natural sources. This method was expected to provide an effective basis for accelerating the development of new hypoglycemic drugs.

在这项工作中，将α-葡萄糖苷酶固定在一种纳米级磁性材料Fe ₃ O ₄ @ ZIF-67上，以构建用于快速筛选α-葡萄糖苷酶抑制剂的生物微反应器。研究了影响固定效率的参数。在最佳条件下，α-葡萄糖苷酶的负载量可达79.07μg·mg ^-1。然后，通过外部磁场将酶生物微反应器固定在两端连接HPLC和微量注射泵的管中，以形成磁性在线筛选系统。优化了筛选过程中的筛选流速和洗脱液有机溶剂比例。对新阳毛尖粗茶提取物进行了测试，以验证该在线筛选方法，并通过该磁性在线筛选系统捕捞了三种抑制剂（儿茶素，表没食子儿茶素没食子酸酯和表儿茶素没食子酸酯）。基于经典pNPG法对α-葡萄糖苷酶的抑制活性进行了进一步的验证和研究。与传统的配体捕捞方法相比，本文建立的筛选方法可以将筛选，洗脱和分析相结合。它可以简单，有效，直接地从天然来源中筛选和鉴定潜在的α-葡萄糖苷酶抑制剂。该方法有望为加速新的降血糖药物的开发提供有效的依据。