Although metal–organic frameworks (MOFs) have been considered as promising matrices for enzyme immobilization, HKUST-1, constructed from copper acetate (CuAc2) and benzene 1,3,5-tricarboxylate (BTC), has rarely been explored for this application. In this study, mushroom tyrosinase (EC 1.14.18.1) was immobilized in the form of tyrosinase@HKUST-1 following a simple reaction procedure by mixing BTC with the enzyme prior to addition of CuAc2. The resultant biocatalyst was characterized in both structural features and catalytic properties. Upon incorporation into the HKUST-1 frameworks, the enzyme gained a prominent enhancement in stability against pH, temperature and storage: When incubated at 50 °C and pH 6.0, tyrosinase@HKUST-1 presented a half-life of 32.6 h, which is 77-fold and over tenfold higher than that of the free enzyme and its other immobilization forms, respectively; and the catalyst fully maintained its activity for at least 2 months when stored at 30 °C. The applicability of this new biocatalyst was demonstrated by employing it as catalyst for regioselective ortho-hydroxylation reactions to produce catecholic products with huge pharmacological effects, i.e., hydroxytyrosol and l-DOPA, with excellent yields and productivities. This study has thus offered a facile immobilization method to prepare a novel biocatalyst with super stability, and tyrosinase@HKUST-1 so formed from crude mushroom extract provides an efficient catalyst which can be applied to the production of catecholic products with health benefits. Graphical Abstract

BACKGROUND Metal–organic frameworks (MOFs) have gained increasing attention with ever‐expanding applications. Developing new MOF‐based immobilized enzymes with new applications is required, not only for demonstrating the generality and applicability of using MOFs for enzyme immobilization, but also to provide potential biocatalysts for industrial applications. To the best of the authors’ knowledge, this is the first report of immobilizing mushroom tyrosinase on zeolitic imidazolate frameworks (ZIFs), with two new applications being developed. RESULTS Upon immobilization through a one‐pot in situ encapsulation approach, the resultant tyrosinase@ZIF‐8 was characterized in structural features and catalytic properties. It was much more stable against pH and temperature relative to the free enzyme. The new biocatalyst was highly efficient in catalyzing the synthesis of catecholic products with pharmacological benefits (l‐DOPA, piceatannol and 3′‐hydroxypterostilbene) and in eliminating phenolic pollutants (phenol, p‐cresol, p‐chlorophenol). Excellent productivities were obtained for the synthesis of the three catecholic products (0.14, 1.38 and 1.46 g L−1 h−1, respectively). A complete removal of the three phenolic pollutants was achieved within 2.5 h, superior to other processes mediated by the same enzyme, or others, immobilized on different supports. The reusability of the new biocatalyst can be remarkably improved by entrapment into polyvinyl alcohol‐alginate gel. CONCLUSIONS Tyrosinase can be immobilized on a new type of MOF with advantages such as easy preparation and excellent catalytic performance. This novel immobilization strategy for tyrosinase offers an excellent biocatalyst potent for both catecholic product synthesis and phenolic wastewater treatment. © 2021 Society of Chemical Industry (SCI).

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Although metal-organic frameworks (MOFs) have been considered as promising matrices for enzyme immobilization, HKUST-1, constructed from copper acetate (CuAc2) and benzene 1,3,5-tricarboxylate (BTC), has rarely been explored for this application. In this study, mushroom tyrosinase (EC 1.14.18.1) was immobilized in the form of tyrosinase@HKUST-1 following a simple reaction procedure we developed. The order of mixing BTC with the enzyme prior to addition of CuAc2 is believed to be responsible for the super stability exhibited by the immobilized enzyme so formed. The resultant biocatalyst was characterized in both structural features and catalytic properties. Upon incorporation into the HKUST-1 frameworks, the enzyme gained a prominent enhancement in stability against pH, temperature and storage: When incubated at 50 oC and pH 6.0, tyrosinase@HKUST-1 presented a half-life of 32.6 h, which is 77-fold and over 10-fold higher than that of the free enzyme and its other immobilization forms, respectively; and the catalyst fully maintained its activity for at least 2 months when stored at 30 oC. The applicability of this new biocatalyst was demonstrated by employing it as catalyst for regioselective ortho-hydroxylation reactions to produce catecholic products with huge pharmacological effects, i.e., hydroxytyrosol and L-DOPA, with excellent yields and productivities. This study has thus offered a facile immobilization method to prepare a novel biocatalyst with super stability, and tyrosinase@HKUST-1 so formed from crude mushroom extract provides an efficient catalyst which can be applied to the production of catecholic products with health benefits.