Abstract
The present study explored the immobilization of laccase onto iron magnetic nanoparticles (MNPs) to enhance its enzymatic properties and applications. The immobilization process was optimized using Box-Behnken design (BBD). BBD showed significance towards the quadratic model with experimental data. Maximum laccase activity recovery (99%) of the predicted model was observed at 0.75 mg/mL of laccase concentration, 200 mg/mL of MNPs, 0.3% cross linking with carbodiimide, and 3 h of cross-linking time. The magnetization activity of MNPs (8 emu/g) and the immobilized laccase with MNPs (4 emu/g) was analyzed using vibrating sample magnetometer (VSM). Maximum activity of immobilized laccase was observed at pH 7.0 and 55 °C. The immobilized laccase has greater stability (100 h) and significant chlorpyrifos (pesticide) degradation activity. High-performance liquid chromatography (HPLC) results confirmed the degraded metabolic products of chlorpyrifos. In all, the immobilized laccase was superior to free laccase, showing promising structural and application characteristics.