Immobilization of penicillin G acylase (PGA) on carrier was an established approach to improve the stability and reusable property of it. Herein, Magnetic Fe3O4 nanoparticles (Fe3O4 NPs) that coated by poly (tannic acid), named as Fe3O4 @PTA nanoparticles (Fe3O4 @PTA NPs), had been used as carrier for immobilizing of PGA by physical adsorption effect to produce Fe3O4 @PTA-PGA NPs. Then Fe3O4 @PTA-PGA NPs was post-modified by spontaneous in-situ self-polymerization of dopamine to prepare polydopamine (PDA) coated Fe3O4 @PTA-PGA/PDA nanocapsule. Fe3O4 @PTA-PGA/PDA nanocapsule with sandwich-like structure was a biomimetic composite penicillin G acylase-based biocatalyst, having excellent operational stability. The response rate of Fe3O4 @PTA-PGA/PDA nanocapsule decreased with the extension of polymerization time, whereas the storage stability, thermal stability and pH stability increased with the extension of polymerization time. The EAR of Fe3O4 @PTA-PGA/PDA nanocapsule was 76.4 % with PDA coating time of 4 h. The relative activity (Rt) of Fe3O4 @PTA-PGA/PDA magnetic nanocapsule was 87.1 %, while that of free PGA was 12.5 % after 90 days of storage. Moreover, the Fe3O4 @PTA-PGA/PDA nanocapsule still had 94.1 % of its initial enzyme activity retained and with a enzyme recovery ratio (Re) of 98.0 % after 12 cycles of repeated usage. All those results indicated that the Fe3O4 @PTA-PGA/PDA nanocapsule had excellent operation stability and reusable property, which was very promising for being used as biocatalyst in industrial applications.
Read full abstract