A biofilter with fungus was developed for efficient degradation of benzene, which can overcome the potential risk of leakage commonly found in such services. Results indicated that the optimum parameter values were temperature 40 °C, pH 6, and 500 mg L-1 of the initial benzene concentration. Besides, the empty bed residence time and inlet load range of biofilter were set to 20 s and 21.23-169.84 g m-3 h-1 respectively. Under these conditions, this biofilter can obtain the maximum removal efficiency of more than 90%, the eliminating capacity could be up to 151.67 g m-3 h-1. Furthermore, scanning electron microscopy was used to investigate three filler materials for packing fungus biofilm. This is the first study introducing an Aspergillus strain for benzene removal and these results highlight that the development of this biofilter has the potential scaling-up application as gas-processing of industrial wastes.