Non-covalent and covalent immobilization of papain onto Ti₃C₂ MXene nanosheets

Papain was immobilized onto Ti₃C₂ MXene nanosheets by physical adsorption and physical adsorption combined with covalent crosslinking with glutaraldehyde. Ti₃C₂ MXene nanosheets were prepared by hydrofluoric acid etching method. The resulting products were well characterized by SEM, BET, XRD, FTIR, XPS. The optimized immobilization conditions are pH 6.5, immobilization time of 20 h, immobilization temperature of 10℃, and 10 mL 2 mg mL^-1 papain, the amount of papain immobilized was 156 mg g^-1, the activity of the immobilized papain determined was 1701 U∙g^-1. The immobilized papain exhibited enhanced pH and temperature endurances, immobilized papain also showed improved storage stability (39.25 % and 65.57 % after 20 days of storage at 4 °C). papain reusability was significantly improved after immobilization and it retained more than 50 % of its initial activity after 5 repeated cycles. Interestingly, the results of immobilized enzymes demonstrated that the immobilization of enzymes on Ti₃C₂ MXene is feasible. Such approach could be transferred to other support systems for anchoring enzyme.