In the present study novel polypyrrole-cellulose-graphene oxide nanocomposite (PCeGONC) was employed for the immobilization of ginger peroxidase (GP) via simple adsorption mechanism. Immobilization of enzyme on the obtained support resulted in enhancement of the enzyme activity. The recovery of activity was 128% of the initial activity. Consequently, in 3 h stirred batch treatment, PCeGONC bound GP exhibited higher decolorization efficiency (99%) for RB4 dye as compared to free GP (88%). The immobilized GP exhibited higher operational stability and retained approximately 72% of its initial activity even after ten sequential cycles of dye decolorization in batch process. The kinetic characterization of PCeGONC bound GP revealed slightly lower K_m and 3.3 times higher V_max compared to free GP. Degraded products were identified on the basis of GC-MS analysis and degradation pathway was proposed accordingly which confirms enzymatic breakdown of RB 4 into low molecular weight compounds. Genotoxic assessment of GP treated RB 4 revealed significant reduction of its genotoxic potential. In-silico analysis identified that binding site of PCeGONC on enzyme is distinct and lies far away from the active site of the enzyme. Furthermore, it also revealed higher affinity of 1-hydroxybenzotriazole (a redox mediator) and RB 4 for PCeGONC bound enzyme as compared to the free enzyme. This is in consensus with the observed decrease in K_m of the immobilized GP.

在本研究中，新型聚吡咯-纤维素-氧化石墨烯纳米复合材料（PCeGONC）通过简单的吸附机制用于生姜过氧化物酶（GP）的固定化。将酶固定在获得的载体上导致酶活性的增强。活性的恢复是初始活性的128％。因此，在3小时搅拌分批处理中，与游离GP（88％）相比，结合PCeGONC的GP对RB4染料显示出更高的脱色效率（99％）。固定的GP表现出更高的操作稳定性，并且即使在分批过程中连续十次将染料脱色之后，仍保留了其初始活性的约72％。与PCeGONC结合的GP的动力学特征表明，K _m稍低，而V _max高3.3倍与免费GP相比。在GC-MS分析的基础上鉴定了降解产物，并据此提出了降解途径，这证实了RB 4被酶法降解为低分子量化合物。GP处理的RB 4的遗传毒性评估表明其遗传毒性潜力显着降低。计算机分析表明，PCeGONC在酶上的结合位点是独特的，并且与酶的活性位点相距很远。此外，与游离酶相比，它还揭示了1-羟基苯并三唑（氧化还原介体）和RB 4对PCeGONC结合的酶具有更高的亲和力。这与观察到的固定GP的K _m降低是一致的。