中文摘要：

以壳聚糖/蒙脱土插层复合物为载体,吸附法制备固定化多酚氧化酶,并以此催化氧化去除水中的苯酚、4-氯苯酚和2,4-二氯苯酚.考察了固定化多酚氧化酶的制备条件,对酚类化合物的催化氧化条件、动力学特性以及固定化酶的重复使用性能.结果表明,固定化多酚氧化酶的最佳制备条件为p H 5.0,酶与载体质量比20 mg·g-1,固定化6 h,所得固定化酶的载酶量为12.12 mg·g-1,每克单位载体酶活为12.76×103U·g-1.固定化多酚氧化酶对酚类化合物的最佳去除条件为：苯酚溶液p H 7.0,温度30℃;4-氯苯酚溶液p H 5.0,温度20℃;2,4-二氯苯酚溶液p H 5.0,温度30℃.在最佳反应条件下,酶与底物质量比为20 mg·mg-1时,固定化多酚氧化酶对苯酚、4-氯苯酚和2,4-二氯苯酚溶液去除率分别为63.6%、85.8%和87.8%.苯酚、4-氯苯酚和2,4-二氯苯酚的米氏常数Km值依次减小,最大反应速率Vmax依次增大,表明固定化酶对2,4-二氯苯酚的亲和力最强,催化速度最快.固定化酶循环使用6次（24 h）后对苯酚、4-氯苯酚和2,4-二氯苯酚的去除率分别为15.7%、24.2%和27.8%.

英文摘要：

Polyphenol oxidase （PPO） was immobilized on chitosan/montmorillonite intercalation composites by adsorption, and then used to remove phenol, 4-chlorophenol and 2,4-dichlorophenol from aqueous solutions. The immobilization conditions, catalytic oxidation conditions of phenolic compounds, dynamics and reusability of immobilized PPO were investigated in detail. Optimal of immobilized PPO was 12.12 mg.g-1 and 12.76×103 U-g-1 ,respectively.Optimal conditions of the removal for phenolic compounds by immobilized PPO were pH 7.0 and 30 ℃ for phenol,pH 5.0 and 20 ℃ for 4-chlorophenol,pH 5.0 and 30 ℃ for 2,4-dichlorophenol.When enzyme/substrate weight ratio was 20 rag·mg-1, the removal efficiency of phenol, 4-chlorophenol and 2,4-dichlorophenol under optimal conditions was 63.6%, 85.8% and 87.8%, respectively.Michaelis constant Km values of phenol,4-ehlorophenol and 2,4-dichlorophenol decreased, while the maximum reaction rate Vmax increased gradually, indicating that 2,4-dichlorophenol possessed the highest affinity for the immobilized enzyme and fastest catalysis rate.The removal efficiency of phenol, 4-chlorophenol and 2,4-dichlorophenol was 15.7%, 24.2% and 27.8%, respectively, after 6 times repeated use of immobilized enzyme.