Comparative study of poly tannic acid functionalized magnetic particles before and after modification for immobilized penicillin G acylase.

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.

Magnetic thermosensitive polymer composite carrier with target spacing for enhancing immobilized enzyme performance.

A novel magnetic thermosensitive polymer composite carrier with target spacing was developed. In this strategy, thermosensitive polymer grafted on magnetic Fe3O4 for enhancing immobilized penicillin G acylase (PGA) performance and introduce immobilized target spacing into magnetic carriers for the first time. Fe3O4 nanoparticles were synthesized by a reverse microemulsion method. The modifier used was the silane coupling agent γ-methylacryloxypropyl trimethoxysilane (KH570) and then reacting with a reversible-adaptive fragmentation chain transfer (RAFT) reagent, 2-cyano-2-propyldodecyl trithiocarbonate (CPDTC). The thermo-sensitive nanoparticle-composite carrier of Fe3O4-grafted-poly N, N-diethyl acrylamide-block-poly ß-Hydroxyethyl methacrylate-block-random copolymer of glycidyl methacrylate and methyl methacrylate (Fe3O4-g-PDEA-b-PHEMA-b-P(MMA-co-GMA)) were synthesized by RAFT polymerization technique that used N, N-diethyl acrylamide (DEA), ß-Hydroxyethyl methacrylate (HEMA), Glycidyl methacrylate (GMA) and Methyl methacrylate (MMA) as monomer, then which were employed as functional carriers for the immobilization of PGA. Within the carrier, the epoxy group of GMA segment was a target immobilization site for PGA and the introduction of MMA reflected the target space of immobilized PGA to improve catalytic activity and catalytic activity recovery rate of the immobilized PGA. Characterizations demonstrated that the triblock copolymers grafted Fe3O4 nanoparticles were successfully fabricated by the structure design. Besides, under these circumstances the enzyme activity (EA), enzyme loading capacity (ELC) and catalytic activity recovery ration (CAR) reached 31235 U/g, 128.39 mg/g and 93.32 %, respectively. The catalytic activity of immobilized PGA maintained 87.4 % of initial value and the recovery ratio (R) of immobilized PGA reached 96.22 % after recycling 12 times. Furthermore, the immobilized PGA exhibited advantages of low temperature homogeneous catalysis and magnetic separation, which indicated broad application prospects in the biocatalysts' field.