Facile Purification and Immobilization of Organophosphorus Hydrolase on Protein-Inorganic Hybrid Phosphate Nanosheets

Oriented immobilization of enzymes helps to maintain their native structure and proper orientation for high-performance engineering to meet extensive biocatalysis demands. However, the supporting materials used for orientated immobilization are usually costly or complicated in preparation, affecting their practical applications. In this work, a facile purification and immobilization method was proposed for enzyme immobilization based on organic–inorganic hybrid calcium phosphate nanocrystal（Ca Ps）induced by Cu2+modified bovine serum albumin（BSA-Cu）. Then, the as-prepared hybrid calcium phosphate nanosheet, BSA-Cu@Ca Ps, was utilized for one-pot purification and immobilization of His-tagged organophosphorus hydrolase（OPH） by metal-affinity binding to the incorporated BSA. BSA-Cu@Ca PsOPH exhibited enhanced p H stability and thermal stability compared to the free enzyme. Moreover,BSA-Cu@Ca Ps-OPH could retain more than 75% and 56% of initial activity after reuse 5 and 10 times,respectively. The results demonstrated that this facile strategy was promising for the effective biodegradation of organophosphorus pesticides with the immobilized enzyme.