Biomimetic dynamic membrane (BDM) has been employed as a promising membrane separation technology regarding water/wastewater treatment (Model pollutant is methylene blue). Given its catalytic function on micro-pollutant removal and fouling control, detailed mechanism for impacts of fabrication method, carriers (CNT and GO) and laccase on the construction of biomimetic layer and enzyme immobilization have not been clear so far. In this work, the BDM performance with various fabrication methods, carriers and laccase were investigated and verified. The BDM fabrication tests demonstrated that BDM with mixed filtration method had better filtration performance (up to 120 L m h flux and 80% removal rate) than BDM with stepwise filtration method. Moreover, the laccases immobilized on GO exhibited a stronger laccase activity than those on CNT. Increasing CNT or GO dosage strengthened removal rate, but lowered flux, meanwhile flux and removal rate exhibited a significant fluctuation with certain laccase dosage. At 25 g m CNT or GO dosage and 50 g m laccase dosage, the optimized flux and removal rate values were obtained. Further study investigated the surface morphology and property of BDM, showing that BDM with mixed filtration method turned out to be the optimized enzyme immobilization mechanism and fabrication method. In addition, during multiple filtration cycles, with the optimized conditions, the removal rate, flux and laccase activity of BDM could maintain at high levels. On account of the finding of the present study, selecting a suitable fabrication method, appropriate CNT or GO dosage and laccase dosage can indeed optimize the structure of biomimetic layer and enzyme immobilization, expanding its possibility on sustainable operation.