Tyrosinase-mediated dopamine polymerization modified magnetic alginate beads for dual-enzymes encapsulation:...

In this study, a simple and efficient method to obtain entrapment of mixtures of double enzymes is developed. As a proof of principle, double enzymes (tyrosinase (TYR) and β-glucosidase (β-Glu)) were co-immobilized in magnetic alginate-polydopamine (PDA) beads using in situ TYR-mediated dopamine polymerization and internal setting strategy-mediated magnetic alginate-PDA gelation. The leakage of enzymes from the magnetic alginate beads was significantly reduced by exploiting the double network cross-linking of alginate and PDA, which was induced by the d-(+)-Gluconic acid δ-lactone (GDL) and TYR, respectively. The physicochemical properties of the prepared magnetic alginate beads were characterized by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and scanning electron microscopy (SEM) analysis. After that, the enzymatic reaction conditions and the performance of the entrapped TYR and β-Glu, such as enzyme kinetics and inhibition kinetics, were investigated. The Michaelis-Menten constants (K_m) of the entrapped TYR and β-Glu were determined as 2.72 and 3.45 mM, respectively. The half-maximal inhibitory concentrations (IC₅₀) of kojic acid and castanospermine for the entrapped TYR and β-Glu were determined as 13.04 and 56.23 μM, respectively. Finally, the entrapped double enzymes magnetic alginate beads were successfully applied to evaluate the inhibitory potency of six kinds of tea polyphenols extracts. Black tea and white tea showed high inhibition activity against TYR were (36.14 ± 1.43)% and (36.76 ± 2.35)%, respectively, while the black tea and dark tea showed high inhibition activity against β-Glu were (37.89 ± 6.70)% and (21.28 ± 4.68)%, respectively.