Abstract:[Objective] To mitigate the threat of heavy metal pollution in wastewater to global food safety and human health, reduce the accumulation of lead (Pb) in soil, plants, and animals, and improve the removal rate of heavy metals by immobilizing microbial strains. [Methods] We carried out mixed strain test to select the white rot fungal strains with strong Pb2+ removal effects and excellent compatibility and explored the optimal strains and ratio for combination. Furthermore, we optimized the formula of the fungal strain composite and explored the optimal adsorption conditions of the composite in application. [Results] Phanerochaete chrysosporium, Coriolus versicolor, Lentinus sajor-caju, and Pleurotus ostreatus with good compatibility were selected for subsequent experiments. C. versicolor mixed with L. sajor-caju at a volume ratio of 1:1 outperformed the single strains in removing Pb2+. The fungal strain composite composed of 20.0 g/L sodium alginate, 15.0 g/L biochar, 2.0×106 CFU/mL white rot fungi, silica, and zeolite showed the Pb2+ removal rate of 90.63% within 96 h. Moreover, this composite had higher mechanical strength and strong resistance to mechanical shear. At the addition amount of 8.35 g/L and pH 5.64, the composite demonstrated the Pb2+ removal rate of 97.45% within 96 h. Moreover, this composite can be reused 7 times after adsorption-desorption-readsorption and maintained high Pb2+ removal capacity. [Conclusion] The immobilized white rot fungal strain composite can significantly improve the microbial utilization rate and wastewater treatment efficiency compared with single strains. It can greatly adsorb Pb2+ in wastewater under appropriate conditions within a short time and reduce the environmental threat caused by heavy metal pollutants. Therefore, the promotion of environmental protection greatly benefits from the use of immobilized mixed white rot fungal strains in the treatment of heavy metal-contaminated wastewater.