Phytase is of great interest for biotechnological applications, in particular for the reduction of phytate content in feed and food industries. The enzyme minimizes the need for supplementation with inorganic phosphorus, due to its ability to improve the utilization of organic phosphorus in poultry and thus markedly reduces the excretion of phosphorus in manure. Two different strains (a' and b') similar to Aspergillus species were isolated from soil by using agar plates for the production of Phytase. Three different media were used for the Phytase production and out of those, wheat bran was found to be most suitable substrate for phytase production by the in comparison to rice bran and corn meal.