Abstract: Enzyme immobilization has been accepted as an efficient technique for improving the stability and recyclability of enzymes. Herein, biomimetic mineralization strategy was employed to achieve the immobilization of urease in a type of metal-organic frameworks (zeolite imidazolate framework-8, ZIF-8), and the immobilized enzyme urease@ZIF-8 was systematically evaluated for its structure, activity, stability and recyclability, using the hydrolysis of urea as a model. The entrapment of urease was found to be realized in a synchronous manner with the formation of ZIF-8 crystal. The loading of urease in ZIF-8 was measured to be ca. 10.6% through the bicinchoninic acid (BCA) protein assay. The encapsulated urease could efficiently maintain its native conformation, which endowed the immobilized urease with excellent activity and stability, even in harsh conditions (e.g., in the presence of trypsin, acidic or alkali conditions, or at high temperature). Further, urease@ ZIF-8 exhibited good recyclability during the degradation of urea, in which it could keep 58.86% of initial activity after being used for 5 cycles. Thus, biomimetic mineralization could be potentially utilized as a promising method to prepare immobilized ureases with superior activity, stability and recyclability, thereby facilitating the construction of efficient catalysts for industrial biocatalysis and biosensing.

Key words: Urease, Metal-organic frameworks, Immobilization, Biomimetic mineralization, Stability, Recyclability

摘要： Enzyme immobilization has been accepted as an efficient technique for improving the stability and recyclability of enzymes. Herein, biomimetic mineralization strategy was employed to achieve the immobilization of urease in a type of metal-organic frameworks (zeolite imidazolate framework-8, ZIF-8), and the immobilized enzyme urease@ZIF-8 was systematically evaluated for its structure, activity, stability and recyclability, using the hydrolysis of urea as a model. The entrapment of urease was found to be realized in a synchronous manner with the formation of ZIF-8 crystal. The loading of urease in ZIF-8 was measured to be ca. 10.6% through the bicinchoninic acid (BCA) protein assay. The encapsulated urease could efficiently maintain its native conformation, which endowed the immobilized urease with excellent activity and stability, even in harsh conditions (e.g., in the presence of trypsin, acidic or alkali conditions, or at high temperature). Further, urease@ ZIF-8 exhibited good recyclability during the degradation of urea, in which it could keep 58.86% of initial activity after being used for 5 cycles. Thus, biomimetic mineralization could be potentially utilized as a promising method to prepare immobilized ureases with superior activity, stability and recyclability, thereby facilitating the construction of efficient catalysts for industrial biocatalysis and biosensing.

关键词: Urease, Metal-organic frameworks, Immobilization, Biomimetic mineralization, Stability, Recyclability