Preparation of phytic acid modified α-Glucosidase/Cu₃(PO₄)₂·3H₂O hybrid nanoflower and its application

A simple, convenient and efficient enzyme immobilization method through phytic acid (PA) modified α-Glucosidase (α-Glu)/Cu₃(PO₄)₂·3H₂O hybrid nanoflower was developed. The structural properties of the materials were studied by several characterization techniques. Subsequently, the enzymatic reaction conditions such as the pH value and temperature were optimized, and the enzyme kinetics and inhibition parameters were determined. The PA modified α-Glu/Cu₃(PO₄)₂·3H₂O hybrid nanoflower had better enzymatic activity under a wide pH range and high temperature than the free one. After seven successive cycles, the PA modified α-Glu/Cu₃(PO₄)₂·3H₂O hybrid nanoflower could still maintain approximately 63.0 % of its initial immobilized enzyme activity. The Michaelis-Menten constant (K_m) and the half-maximal inhibitory concentration (IC₅₀) of acarbose were determined as 0.77 mM and 15.01 μM, respectively. In addition, the material was applied to evaluate the inhibitory activity of ten phenolic compounds on α-Glu, and epicatechin gallate, gallocatechin gallate, epigallocatechin gallate and rosmarinic acid showed good inhibitory activity with % of inhibition of (53.42 ± 2.39)%, (37.28 ± 1.32)%, (37.08 ± 0.63)% and (35.53 ± 0.23)%, respectively. These results indicate that the PA modified hybrid nanoflower is an efficient method of α-Glu immobilization.