School of Pharmacy, Lanzhou University, Lanzhou 730000, China.
College of Pharmacy, Gansu University of Chinese Medicine, Lanzhou 730000, China.
Langmuir. 2023 Apr 18;39(15):5239-5249. doi: 10.1021/acs.langmuir.2c02979. Epub 2023 Apr 4.
The technology based on immobilized enzymes was employed to screen the constituents inhibiting disease-related enzyme activity from traditional Chinese medicine, which is expected to become an important approach of innovative drug development. Herein, the FeO@POP composite with a core-shell structure was constructed for the first time with FeO magnetic nanoparticles as the core, 1,3,5-tris (4-aminophenyl) benzene (TAPB) and 2,5-divinylterephthalaldehyde (DVA) as organic monomers, and used as the support for immobilizing α-glucosidase. FeO@POP was characterized by transmission electron microscopy, energy-dispersive spectrometry, Fourier transform infrared, powder X-ray diffraction, X-ray photoelectron spectroscopy, and vibrating sample magnetometry. FeO@POP exhibited a distinct core-shell structure and excellent magnetic response (45.2 emu g). α-Glucosidase was covalently immobilized on core-shell FeO@POP magnetic nanoparticles using glutaraldehyde as the cross-linking agent. The immobilized α-glucosidase possessed improved pH stability and thermal stability as well as good storage stability and reusability. More importantly, the immobilized enzyme exhibited a lower value and enhanced affinity for the substrate than the free one. The immobilized α-glucosidase was subsequently used for inhibitor screening from 18 traditional Chinese medicines in combination with capillary electrophoresis analysis among which exhibited the highest enzyme inhibitory activity. These positive results demonstrated that such magnetic POP-based core-shell nanoparticles were a promising carrier for enzyme immobilization and the screening strategy based on immobilized enzyme provided an effective way to rapidly explore the targeted active compounds from medicinal plants.
基于固定化酶的技术被用于筛选中药中抑制疾病相关酶活性的成分，有望成为创新药物开发的重要途径。本文首次构建了具有核壳结构的 FeO@POP 复合材料，以 FeO 磁性纳米粒子为核，1,3,5-三（4-氨基苯基）苯（TAPB）和 2,5-二乙烯基对苯二甲醛（DVA）为有机单体，作为固定α-葡萄糖苷酶的载体。通过透射电子显微镜、能谱、傅里叶变换红外光谱、粉末 X 射线衍射、X 射线光电子能谱和振动样品磁强计对 FeO@POP 进行了表征。FeO@POP 表现出明显的核壳结构和优异的磁响应（45.2 emu g）。用戊二醛作为交联剂，将α-葡萄糖苷酶通过共价键固定在核壳 FeO@POP 磁性纳米粒子上。固定化α-葡萄糖苷酶具有更好的 pH 稳定性和热稳定性，以及良好的储存稳定性和可重复使用性。更重要的是，固定化酶对底物的亲和力和米氏常数（Km 值）均低于游离酶。随后，将固定化酶与毛细管电泳分析相结合，用于从 18 种中药中筛选抑制剂，其中 表现出最高的酶抑制活性。这些积极的结果表明，这种基于磁性 POP 的核壳纳米粒子是一种很有前途的酶固定化载体，基于固定化酶的筛选策略为从药用植物中快速探索靶向活性化合物提供了一种有效方法。
