In this work, Fe3O4 nanoparticles (NPs) was synthesized by inverting microemulsion method. After that, based on the physical and chemical properties of tannic acid (TA), poly tannic acid (PTA) was coated on Fe3O4 NPs surface. Fe3O4 NPs coated with PTA, on the one hand, was used to immobilize Penicillin G acylase (PGA) by physical adsorption. On the other hand, it was modified by glutaraldehyde (GA). GA grafting rate (Gr-GA) was optimized, and the Gr-GA was 30.0% under the optimum conditions. Then, through the Schiff base reaction between the glutaraldehyde group and PGA amino group, this covalent immobilization of PGA was further realized under mild conditions. Finally, the structures of every stage of magnetic composites were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), vibration magnetometer (VSM) and transmission electron microscopy (TEM), respectively. The results indicated that the enzyme activity (EA), enzyme activity recovery (EAR) and maximum load (ELC) of the immobilized PGA were 26843 U/g, 80.2% and 125 mg/g, respectively. Compared to the physical immobilization of PGA by only coating PTA nanoparticles, further modified nanoparticles by GA showed higher catalytic stability, reusability and storage stability.
Keywords: Fe3O4 nanoparticles; covalent immobilization; glutaraldehyde; physical adsorption; poly tannic acid.