Immobilization of Zanthoxylum armatum peroxidase and its application for phenol degradation in aqueous systems

This study pioneers the covalent immobilization of Zanthoxylum armatum peroxidase (ZADTP) onto magnetic agarose phenylboronic acid microspheres (MAM@PA) for phenol-contaminated water remediation. Under optimized immobilization conditions (pH 8.0, 25°C, enzyme concentration 1.0 mg/mL, 90 min), the system achieved a loading capacity of 38.50 mg/g with 80.21 % activity retention. Characterization via SEM, FTIR, XRD, and VSM confirmed uniform enzyme distribution on the carrier surface without disrupting its crystalline structure, while superparamagnetic properties enabled efficient magnetic separation. Compared to free ZADTP, the immobilized enzyme demonstrated enhanced stability across broad pH (5.0–8.0) and temperature (20–70°C) ranges, superior storage stability, and strong tolerance to metal ions. It retained 78 % activity after 5 reuse cycles and 68 % activity post 50-day storage at 4°C. Catalytic evaluations revealed phenol degradation efficiencies of 93.67 % (2 mM) and 81.67 % (10 mM), with post-treatment solutions exhibiting significantly reduced cytotoxicity (LO2 cell viability: 97.66 %). In conclusion, the ZADTP@MAM@PA immobilized enzyme demonstrates great potential for application in water remediation and ecological environment improvement.