Exopolysaccharides (EPS) are bioactive metabolites with high molecular weight and are produced by several microorganisms such as endophytic fungi. Thus, the aim of the present study was designed to explore the biosynthesis and characteristics of EPS by the endophytic fungus Neopestalotiopsis clavispora AUMC15969 and then investigate some environmental and nutritional factors that affect their production. Moreover, we estimated an additional value, namely the production of bio diesel. Maximum production of EPS was 7.86 g/L when N. clavispora was grown on lactose as the sole carbon source and peptone as the nitrogen source, respectively, and pH 7 at 35 °Cfor 10 days. The produced EPS had a total sugar content of 0.93 g/g where protein content was 0.076 g/g. It revealed a strong antioxidant activity that improved with increasing sample concentration, with the optimum concentration of 10 mg/mL producing 83.1% DPPH radical scavenging activity with an IC50 equal to 3.89 mg/mL. The extracted lipid from the fungal mycelia at the end of the fermentation process was 31.76% w/w. The biodiesel produced from the transesterification of lipids was 87.4% total fatty acid methyl esters. The present study demonstrated the potential production of EPS and lipid biopolymers in one-pot fermentation which could use as a resource for industrial technologies