In the current study, an experimental design was performed on fresh water microalgae, Scenedesmus obliquusconsidering initial nitrogen, phosphorus and iron concentration as independent variables using Box Behnken design of RSM.Various physio-biochemical componentswere studied under varied concentration of nutrients in BG-11(mgmL-1) media, Nitrate,10-750, Phosphate, 40-120, Ferric ammonium citrate, 3-9 with optimized lipid yield of 55.52 % (dcw) and biomass yield 4.89gL-1 were achieved, which confirmed the validity of the predicted model.Prob>F value of <0.05 and >0.05 indicated model terms which were significant and non-significant, respectively. The fit of the model yielded R2 values up to 96.25%, 94.12% for lipids, biomass respectively. GCMS study of Scenedesmus obliquus, FAME profile of Scenedesmus obliquus profile contained Palmitic (C16:0, 52.10% stearic acid (C18:0, 17.21%), Oleic (C18:1, 12.60%), Linoleic C18:2 (10.01%), and Linolenic (C18:3, 12.70%) whichshowed that Scenedesmus obliquus possessed a favourable fatty acids profile that can be utilized for biodiesel production.