Immobilization of polyphenol oxidase on chitosan/organic rectorite composites for phenolic compounds removal

Chitosan/organic rectorite (CTS/OREC) composites were prepared and characterized by Fourier transform infrared spectrometry and X-ray diffraction. Polyphenol oxidase (PPO) was immobilized on CTS/OREC by physical adsorption (APPO) and covalent binding (CPPO). Taguchi method was applied in the optimization of immobilization conditions resulting in the highest enzyme activity of 16.37 × 10³ and 8.92 × 10³U/g for APPO and CPPO, respectively. APPO enzyme activity was higher than that of CPPO, while CPPO showed the higher enzyme loading capacity than that of APPO. The removal percentage of phenolic compound, including phenol (PH), 4-chlorophenol (4-CP) and 2,4-dichlorophenol (2,4-DCP), by immobilized PPO was also explored. The results indicated that APPO was more efficient in phenolic compounds removal than CPPO. APPO contributed to a quick removal in the first hour, and the removal percentage of PH, 4-CP and 2,4-DCP could reach 69.3 ± 4.2%, 89.8 ± 2.5% and 93.8 ± 1.7% within 2 h, respectively. The order of removal percentage of phenolic compounds for both immobilized PPO was 2,4-DCP > 4-CP > PH. After 10 consecutive operations, the removal percentage of 2,4-DCP reached 73.2 ± 2.6% and 60.3 ± 1.5% for APPO and CPPO, respectively. The results introduced a novel support for PPO immobilization, and the immobilized PPO had great potential in wastewater treatment.