Functionalization of chitosan by metformin, nickel metal ions and magnetic nanoparticles as a nanobiocomposite for purification of alkaline phosphatase from hen's egg yolk.

In this study, a novel and green chitosan-metformin/NiCl/FeO nanobiocomposite was synthesized and used to purify alkaline phosphatase (ALPs) from hen's egg yolk. For this purpose, after functionalization of the chitosan biopolymer by terephthaloyl chloride-metformin ligand, the coordination with Ni(II) and magnetization process were performed. The structure and properties of the synthesized nanobiocomposite were then evaluated by using analyzes such as FT-IR, EDX, FE-SEM, XRD, TGA and VSM. Purification of ALPs with chitosan-metformin/NiCl/FeO nanobiocomposite is a fast, reusable and cost-effective method. By this protocol, 62% purification efficiency was obtained and the synthesized nanobiocomposite was not attached to other proteins in hen's egg yolk. ALPs was obtained approximately in the pure form and the purification process was evaluated using SDS-PAGE. The reusability of nanobiocomposites was evaluated and a slight decrease in adsorption capacity was observed after 4 cycles.