Facile purification and immobilization of organophosphorus hydrolase on protein-inorganic hybrid phosphate nanosheets

doi:10.1016/j.cjche.2022.07.017

Abstract

Abstract: 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 (CaPs) induced by Cu²⁺ modified bovine serum albumin (BSA-Cu). Then, the as-prepared hybrid calcium phosphate nanosheet, BSA-Cu@CaPs, was utilized for one-pot purification and immobilization of His-tagged organophosphorus hydrolase (OPH) by metal-affinity binding to the incorporated BSA. BSA-Cu@CaPs-OPH exhibited enhanced pH stability and thermal stability compared to the free enzyme. Moreover, BSA-Cu@CaPs-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.

Key words: Calcium phosphate nanocrystal (CaPs), Biomimetic mineralization, Organophosphorus hydrolase (OPH), Oriented immobilization, Hydrolysis, Methyl parathion

摘要： 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 (CaPs) induced by Cu²⁺ modified bovine serum albumin (BSA-Cu). Then, the as-prepared hybrid calcium phosphate nanosheet, BSA-Cu@CaPs, was utilized for one-pot purification and immobilization of His-tagged organophosphorus hydrolase (OPH) by metal-affinity binding to the incorporated BSA. BSA-Cu@CaPs-OPH exhibited enhanced pH stability and thermal stability compared to the free enzyme. Moreover, BSA-Cu@CaPs-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.

关键词: Calcium phosphate nanocrystal (CaPs), Biomimetic mineralization, Organophosphorus hydrolase (OPH), Oriented immobilization, Hydrolysis, Methyl parathion

Zhenfu Wang, Jie Gao, Qinghong Shi, Xiaoyan Dong, Yan Sun. Facile purification and immobilization of organophosphorus hydrolase on protein-inorganic hybrid phosphate nanosheets[J]. Chinese Journal of Chemical Engineering, 2023, 56(4): 119-125.

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