α-glucosidase immobilization on magnetic core-shell metal-organic frameworks for inhibitor screening from traditional Chinese medicines.

In this work, a simple and rapid screening strategy was developed combining capillary electrophoresis analysis with enzymatic assay based on immobilized α-glucosidase. For α-glucosidase immobilization, magnetic core-shell metal-organic frameworks composite (FeO@CS@ZIF-8) was fabricated by a step-by-step assembly method, and α-glucosidase was in situ encapsulated in crystal lattice of ZIF-8. The composite was characterized by transmission electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction and vibrating sample magnetometer. After immobilization, α-glucosidase exhibited enhanced tolerance to temperature and pH, and its reusability was greatly improved with 74 % of initial enzyme activity after being recycled 10 times. The Michaelis-Menten constant of immobilized enzyme was calculated to be 0.47 mM and its inhibition constant and IC for acarbose were 0.57 μM and 0.18 μM, respectively. The immobilized enzyme was subsequently applied to inhibitor screening from 14 TCMs, and Rhei Radix et Rhizoma was screened out. Among the commercially available 10 components presented in Rhei Radix et Rhizoma, gallic acid, (+)-catechin and epicatechin exhibited the strongest inhibitory effect on α-glucosidase. Their binding sites and modes with α-glucosidase were simulated via molecular docking to further verify the inhibition screening assay results. The positive results indicated that the FeO@CS@ZIF-8-based screening strategy may provide a new avenue for discovering enzyme inhibitors from TCMs.