A silica-chitosan support was employed for laccase immobilization. The hybrid support was obtained using calcium ion as linking agent that coordinates silanol and hydroxyl groups of chitosan. The insoluble biocatalyst was then packed in a column and used in a flow system for phenol removal. The immobilized enzyme reactor (IMER) showed a good storage stability (70 % of activity in 70 days) and good reusability (90−50 % of catalytic activity at the 4th reuse in function of chitosan type). The best performance for the phenol removal was obtained with a low molecular weight chitosan from crab shells at pH 5 and with a flow rate of 0.7 mL/min. The apparent Michaelis–Menten (V_max^app, K_m^app) and the inherent (V_max^inh, K_m^inh) constants were also determined to evaluate the influence of the phenol structure on the performance of the system. The enzymatic oxidation of a phenol mixture (4-methylcatechol, catechol, caffeic acid, syringic acid, vanillic acid, p-coumaric acid, and tyrosol) was followed for 21 h in a continuous mode by HPLC. The phenol mixture removal of 90 % was also confirmed by Folin‐Ciocalteu assay.

使用二氧化硅-壳聚糖载体固定漆酶。使用钙离子作为连接剂来配位壳聚糖的硅烷醇和羟基，获得混合载体。然后将不溶性生物催化剂装入柱中并用于流动系统以去除苯酚。固定化酶反应器 (IMER) 显示出良好的储存稳定性（70 天内活性的 70%）和良好的重复使用性（壳聚糖类型功能的第 4 次重复使用时催化活性的 90-50%）。使用来自蟹壳的低分子量壳聚糖在 pH 值为 5 和流速为 0.7 mL/min 的情况下，去除苯酚的性能最佳。表观 Michaelis-Menten (V _max ^app , K _m ^app ) 和固有 (V _max ^inh , K_m ^inh ) 常数也被确定以评估苯酚结构对系统性能的影响。通过 HPLC 以连续模式跟踪苯酚混合物（4-甲基儿茶酚、儿茶酚、咖啡酸、丁香酸、香草酸、对香豆酸和酪醇）的酶促氧化反应21 小时。Folin-Ciocalteu 测定法也证实了苯酚混合物的去除率为 90%。