Tannic acid-aminopropyltriethoxysilane co-deposition modified polymer membrane for α-glucosidase immobilization

Mussel-inspired catechol-amine co-deposition is an effective modification strategy for various materials. In this work, polyvinylidene fluoride (PVDF) membrane was employed as the carrier for α-glucosidase immobilization. By virtue of the co-polymerization of tannic acid (TA) and 3-aminopropyltriethoxysilane (APTES) and the hydrolysis of APTES, a hierarchical layer-colloidal nanospheres coating was decorated on the surface of PVDF membrane. Subsequently, α-glucosidase was covalently bound to the modified PVDF membrane through Schiff base reaction and Michael addition reaction between the residual quinine groups in the coating and the amino groups in enzyme molecules. Several parameters affecting the immobilization procedure were investigated thoroughly. The morphology and functional groups of the prepared composite were characterized by scanning electron microscopy (SEM), Fourier transform infrared-Attenuated total reflectance spectroscopy (FTIR-ATR), and X-ray photoelectron spectroscopy (XPS). Combined with capillary electrophoresis (CE) analysis, the performance, enzyme reaction kinetics and inhibition kinetics of PVDF-immobilized α-glucosidase were studied. The immobilized enzyme exhibited the enhanced tolerance to temperature and pH value. In addition, it possessed good reusability maintaining 77.1% of initial relative activity after 11 recycles, and batch-to-batch reproducibility with RSD of 4.3% (n = 10). The Michaelis-Menten constant (K_m) of immobilized enzyme was calculated to be 4.16 mM, and IC₅₀ value of acarbose was 0.10 µM. Finally, the PVDF-immobilized α-glucosidase was applied to screening potential inhibitors from 13 kinds of traditional Chinese medicines (TCMs), among which Sanguisorba Radix exhibited the strongest inhibitory activity. The positive results suggested that TA/APTES co-deposition was a simple and mild functionalization method for chemically inert polymer membrane and the proposed screening method was a reliable approach for discovering enzyme inhibitors from TCMs.