A one-step method for laccase production and immobilization using the waste culture of Ganoderma lucidum (G. lucidum) was developed, and the laccase was immobilized by the mycelium, which was in the form of a white membrane consisting of superfine fibers. This medical and edible membrane was characterized by scanning electron microscope (SEM), and the fiber diameter was found to be between 1 and 3 μm, with a porous structure formed in the membrane. Fourier transform infrared spectroscopy (FT-IR) and thermogravimetric analysis (TGA) showed that the membrane contained polysaccharide groups and had good thermal resistivity. The membrane was used for the decolorization of methyl violet and malachite green, while the MTT test showed that the membrane had good biocompatibility. The experimental results indicated that the membrane might be applicable for other environmental protection applications in the future.