Chlorpyrifos is organophosphorus pesticide widely used in cultivated fields of Egypt. Chlorpyrifos resides are persisted in environment for unpredictable time and threaten public health. Adequate measures to remove chlorpyrifos contamination are essential to minimize environmental pollution and reduce public health hazards. The vital role of soil fungi in degradation of organic matter prompted us to attempt biodegradation of chlorpyrifos using these fungi. This study aims to isolate biodegradable soil fungi capable of metabolizing chlorpyrifos. 5 of 13 fungal strains isolated form chlorpyrifos contaminated soils were able to degrade chlorpyrifos and the results were confirmed by broth assay. Based on their chlorpyrifos tolerance, five fungal strains identified as Trichoderma viride, T. harzianum, Aspergillus niger, A. oryzae and Penicillium citrinum were detected and confirmed as chlorpyrifos biodegradable fungal strains. T. viride was the most effective biodegradable fungi recording a highest dissipation of chlorpyrifos 70.61% while A. niger, T. harzianum followed by A. oryzae were moderate in their biodegradable activities reducing chlorpyrifos to 63.96, 60.31 and 50.79% respectively. On contrast, P citrinum was showed the lowest degradation rate 25.93%, after 15 days of incubation. Results indicated that these tolerant fungi are promising for potentially effective and environmentally safe of chlorpyrifos biodegradation