Use of tyrosinase-inorganic salt hybrid nanoflowers and tyrosinase-MOF hybrid composites for elimination of phenolic pollutants from industrial wastewaters.

Removal of phenolic pollutants from industrial wastewaters is always an important practical problem. Use of enzymes for dephenolization provides a green solution. In this work, enzymatic methods were developed by employing mushroom tyrosinase immobilized as enzyme-Cu3(PO4)2 hybrid nanoflowers and enzyme-metal organic framework (i.e., ZIF-8 and HKUST-1) hybrid composites, which were shown to be superior to processes mediated by tyrosinase immobilized on other supports in both dephenolization efficiency and reusability. Comparatively, tyrosinase@Cu3(PO4)2 and tyrosinase@HKUST-1 were better than tyrosinase@ZIF-8 in both specific activity and dephenolization efficiency. Typical phenolic pollutants, including 3 monophenols (phenol, p-cresol, p-chlorophenol) and 3 bisphenols (BPA, BPB, BPF), can be completely eliminated within 0.5-4 h. The dephenolization order was discussed based on the enzyme's substrate specificity. The operability and reusability of these hybrid biocomposites were highly improved by entrapping into alginate gels or by incorporating with modified magnetic Fe3O4 nanoparticles. Particularly, the magnetic biocatalyst was prepared via a facile one-pot/one-step de novo synthetic strategy, optimized by using response surface methodology (RSM). The as-prepared magnetic tyrosinase@mHKUST-1 retained a high dephenolization efficiency of 81% after 10 cycles and was effective for continuous dephenolization for at least 24 h. These hybrid biocomposites were also successfully applied to treatment of real industrial wastewater from a coke plant.

Tyrosinase@HKUST-1: a super stable biocatalyst efficient for catecholic product synthesis.

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.