Fabrication of immobilized bromelain using cobalt phosphate material prepared in deep eutectic solvent as carrier

The aim of the present work is to fabricate immobilized bromelain based on the specific interaction between the cobalt ions of carrier and the inherent cysteines contained in bromelain molecules. The cobalt phosphate material was prepared as solid support by using choline chloride (ChCl)/betaine-glycerol deep eutectic solvent (DES) as solvent and template for the first time. The Co-P material with lamellate-based structure obtained in the ChCl-glycerol DES at the Co/P ratio of 3:2 showed the best performance for the immobilization of bromelain. The specific interaction between Co²⁺ and bromelain promoted the aggregation of lamellar Co-P, forming flower-like Co-P@bromelain particles. Under the optimum immobilization conditions, the specific enzyme activity of the immobilized enzyme reached the maximum of 71244 U/g. Compared with Co₃(PO₄)₂ prepared in water system, the obtained Co-P@bromelain using the Co-P material synthesized in the ChCl-glycerol DES as carrier exhibited excellent structure stability. In addition, the immobilized Co-P@bromelain also showed higher catalytic efficiency than free bromelain.