Hydrolysis of Magnetic Amylase-Imprinted Poly(ethylene-co-vinyl alcohol) Composite Nanoparticles

Molecularly imprinted polymers (MIPs) have frequently been employed as recognition elements in sensing applications, or for the controlled delivery of small molecule drugs. An equally important but less well studied application is the use of MIPs in the binding and immobilization of active enzymes. In this study, magnetic MIPs (MMIPs) recognizing the enzyme amylase were prepared using phase inversion of poly(ethylene-co-vinyl alcohol) (EVAL) solutions with 27-44 mol % ethylene in the presence of amylase. The size distribution, specific surface area, magnetization, and composition were characterized by dynamic light scattering (DLS), Brunauer-Emmett-Teller (BET) analysis, superconducting quantum interference devices (SQUID), and X-ray diffraction (XRD), respectively. The mean size of MMIPs was ~100 nm and the magnetization was 14.8 emu/g. The activities of both bound template and rebound enzyme was established by measuring glucose production via starch hydrolysis, at different temperatures, for MIPs with different compositions (wt % EVALs and mol % ethylene). The highest hydrolysis activity of MMIPs (obtained with 32 mol % ethylene) was found to be 1545.2 U/g. Finally, compared to the conventional catalysis process, MMIPs have the advantages of high surface area, suspension, easy removal from reaction, and rapid reload of enzyme. The good activity of amylase MMIPs persists after 50 cycles of starch hydrolysis.