Process optimization of polyphenol oxidase immobilization: Isotherm, kinetic, thermodynamic and removal of phenolic...

Chitosan/montmorillonite (CTS/MMT) and chitosan‑gold nanoparticles/montmorillonite (CTS-Au/MMT) composites were prepared, characterized through Fourier transformed infrared (FT-IR), X-ray powder diffraction (XRD), and scanning electron microscopy (SEM), and utilized as support for immobilization of polyphenol oxidase (PPO). PPO was immobilized on CTS/MMT (IPPO) and CTS-Au/MMT (IPPO-Au) by physical adsorption, respectively. In order to achieve simultaneous maximization of immobilization efficiency and enzyme activity, the immobilization process parameters were optimized by Taguchi-Grey relational analysis (TGRA) approach. Under the optimal immobilization condition, the immobilization efficiency and enzyme activity reached at 50.16% and 1.46 × 10⁴ U/mg for IPPO, and 63.35% and 3.01 × 10⁴ U/mg for IPPO-Au, respectively. The isotherm, kinetic and thermodynamics of PPO adsorption were investigated in detail. The adsorption process was better explained by Toth isotherm and Fractal-like pseudo second order model, respectively. Intra-particle diffusion and film diffusion were involved in the adsorption process and intra-particle diffusion was not the only rate-controlling step. The adsorption of PPO was exothermic, physical and spontaneous at the investigated temperature range. The immobilized PPO were used to oxidize phenolic compounds. All investigated phenolic compounds showed the higher conversion as catalyzed by IPPO-Au. For both IPPO and IPPO-Au, the conversion of substituted phenols was higher than that of phenol.