Extracellular polymeric substances are present abundantly in the surroundings which are produced by different types of microorganisms like bacteria, fungi, algae are also produced by different varieties of plant species. Exopolysaccharides (EPS) are high-molecular-weight polysaccharides secreted by microorganisms into the surrounding environment. These polysaccharides have tremendous application in pharmaceutical industries. For the past two decades, exopolysaccharides of fungus have been known to have high-value biomacromolecules. In most of the studies, sugar compositions of fungus Exopolysaccharaide and variation in molecular weight depend upon culture medium composition and different physical conditions that take place during the process of fermentation. In this current study, the selection of nutrients for the EPS production from Curvularia lunata under submerged batch-culture conditions was made using the Plackett - Burman design matrix method. Subsequently, the optimal condition was determined using Response Surface Methodology (RSM). Thus, produced EPS was characterized by GC-MS and thermogravimetric analysis. It was found to have glucose and mannitol and also showed good thermal stability. EPS was subjected to DPPH assay for analyzing antioxidant activity. Free radical scavenging activity was observed by subjecting the sample to the ABTS method and anticancer activities were tested against HeLa cells. EPS was found to be having a good antioxidant activity, Free radical scavenging activity and showed effective anticancer activity against HeLa cells.