The excessive use of pretilachlor (a chloroacetamide herbicide) has raised concern throughout the world as it has been reported as highly toxic. The present study deals with isolating and screening pretilachlor degrading fungal strains. The strains Aspergillus ficuum (AJN2) and Aspergillus sp. (PDF1) isolated using enrichment technique were able to degrade 79% and 73% of pretilachlor respectively as analyzed using HPLC. Further, the immobilization technique was used in the study the pretilachlor degradation ability of the isolated strains. The immobilized spores of the strains AJN2 and PDF1 mineralized 84% and 95% of pretilachlor respectively. The degradation dynamics study revealed that the DT50 value of the herbicide was reduced to 2.4 d in aqueous medium due to the enhanced enzymatic activity. The enzymatic study showed high lignin peroxidase and manganese peroxidase activity by the strains AJN2 and PDF1 respectively. The study confirmed the efficient degradation of pretilachlor by Aspergillus ficuum (AJN2).