This study aimed to extract and characterize phytase from Rhizomucor pusillus, a thermophilic fungus isolated from various soil samples, and investigate its antimicrobial activity. Six different fungal strains were isolated from soil samples at Mansoura University using standard media. These strains were then characterized and screened to determine whether they could produce phytase through submerged fermentation in a 250 ml flask with 1 g of wheat bran and one hundred milliliters of basal medium with the subsequent composition (g/L): 0.5 KCl, 0.1 MnSO4, 0.5 NaCl, 0.2 MgSO4.7H2O, 0.0075 CaCl.6H2O, 1.0 yeast extract, pH (5.0), and autoclaved for 30 minutes at 121 º C. After cooling, the mixture was inoculated with colonies. Following inoculation, the contents of each flask were combined. At 37º C, the flasks were incubated. To get rid of the debris, the flask contents were centrifuged. The phytase activity was measured using the crude supernatant. When thermophilic fungal strains were used to screen the isolated fungi for phytase production, Rhizomucor pusillus showed the highest activity. The second day of high Activity was ascertained through the measurement of optical density and the enzyme activity during the incubation period at pH 5.0 and 37ᵒC. Rhizomucor pusillus can be further utilized for the commercial synthesis of phytase as it was isolated from soil samples exhibiting notable phytase activity. Phytase is an antibacterial agent that works well.