Extracellular laccase from Trametes versicolor IBL-04 was insolubilized in the form of cross-linked enzyme aggregates (CLEAs) for degradation of different recalcitrant dyes. Among various precipitants, polyethylene glycol (PEG) ensured the best activity recovery (72.85%) of laccase-immobilized cross-linked enzyme aggregates (Lac-CLEAs) following ammonium sulfate (63.24%), n-propanol (46.98%), tert-butanol (22.94%), and acetone (14.5%). Lac-CLEAs were catalytically more active and stable over a broader pH range as compared to the free counterpart. Free enzyme lost 70% of its relative activity at 75 °C, whereas the immobilized derivatives, i.e. propanol-Lac-CLEAs, AS-Lac-CLEAs, and PEG-Lac-CLEAs retained 62.69%, 68.48%, and 86.29% of their relative activities, respectively, at this temperature. Though Lac-CLEAs exhibited a reduced affinity towards its substrate, ${\mathit{V}}_{\max }$ values were markedly increased compared to the free enzyme. Lac-CLEAs catalyzed 94.57%, 90.63%, and 78.72% degradation of Remazol Brilliant Blue R, crystal violet, and Reactive Black 5, respectively. A dramatic decline in the peaks after treatment confirms the Lac-CLEAs potential to remove dyes. Interestingly, the immobilized system retained more than 60% of degradation efficiency even after eight successive uses. In conclusion, the present findings suggest the potential of Lac-CLEAs to mitigate a wide range of dyes and emerging contaminants.

Trametes versicolor的细胞外漆酶IBL-04以交联酶聚集体（CLEA）的形式不溶，用于降解不同的顽固染料。在各种沉淀剂中，聚乙二醇（PEG）可确保在硫酸铵（63.24％），正丙醇（46.98％），叔-叔丁基环己基甲酸酯固定的漆酶固定的交联酶聚集体（Lac-CLEAs）的最佳活性回收率（72.85％）丁醇（22.94％）和丙酮（14.5％）。与游离对应物相比，Lac-CLEA在更宽的pH范围内具有更高的催化活性和稳定性。游离酶在75°C时失去了70％的相对活性，而固定化的衍生物（即丙醇-Lac-CLEA，AS-Lac-CLEA和PEG-Lac-CLEA）保留了它们的62.69％，68.48％和86.29％在此温度下的相对活动。尽管Lac-CLEAs对底物的亲和力降低，${\mathit{V}}_{\mathrm{最高}}$与游离酶相比，该值显着增加。Lac-CLEAs分别催化Remazol亮蓝R，结晶紫和活性黑5降解94.57％，90.63％和78.72％。处理后峰的急剧下降证实了Lac-CLEAs具有去除染料的潜力。有趣的是，即使连续使用八次，固定化系统仍能保持60％以上的降解效率。总之，本研究结果表明，Lac-CLEAs可以减轻多种染料和新兴污染物的潜力。