Biosynthesis of kojic acid by fungi has been acquired huge attention nowadays because it can be used as an alternative method to chemically synthesized compounds. The fundamental feature of the microbiological agitation practice was to obtain potential fungi to achieve greater amount of kojic acid, the fungal isolate used in this study exhibited maximum growth on PDA and was identified as A.flavus based on its microscopic and macroscopic characteristics and ITS sequence analysis. In this vignette, appropriate cultivation conditions were carried out to gain a highest quantity of kojic acid. The maximum yield of kojic acid (31g/l) was reached by formulating the medium. The massive amount of kojic acid and maximum biomass was achieved from A. flavus by utilizing yeast extract as a nitrogen source and carbon source as glucose. The optimum incubation temperature employed was at 30^oC, optimal pH was 3. Antagonistic activity was analyzed against both bacterial and fungal pathogens. The ethyl acetate extract exhibit increased antibacterial activity and maximum antifungal activity comparable with methanol, and chloroform.