L-asparaginase is an important enzyme used in the pharmaceutical and food industry and can be produced by wide variety of microorganisms using affordable agro-based materials. The current study aimed to produce L-asparaginase by fungal isolates, which were screened from soil and optimize the production from the most effective fungus. The data showed that among the 7 tested isolates fungus (F-S3) gave the highest L-asparaginase yield, which was completely identified based on the cultural, morphological and molecular properties as Fusarium oxysporum F-S3. This strain was applied on the solid-state fermentation using 7 agro-based materials (peels of orange, pea, potato, banana, lemon, pomegranate and tea waste) for L-asparaginase production. Results revealed that the pomegranate peel has promoted maximum enzyme production. Our study revealed that when pomegranate peel was treated with gamma irradiation at 15.0 kGy for decontamination, the undesired microbial growth was totally inactivated and L-asparaginase production was increased 1.45-fold compared to the pomegranate peel sterilized by autoclave. Optimization of the solid-state fermentation parameters using the one factor at a time technique resulted in maximum L-asparaginase production (280.4 U/gds) was observed using the irradiated pomegranate peel that was supplemented with 0.2 % (w/w) glucose, 1.0 % (w/w) ammonium chloride, L-asparagine 0.6% (w/w), 70% (v/w) initial moisture content, pH was adjusted at 7.0 and inoculated with 3% inoculum volume (107 spores/ml) for 96 h of incubation at 30°C. Therefore, results concluded that the production of L-asparaginase by the F-S3 strain after optimization of the solid-state fermentation parameters was significantly increased by about 1.74-Fold compared to the production before irradiation and optimization.