In this study, a novel microalgal strain, Desmodesmus sp. KAERI-NJ5, was isolated, identified, and evaluated as a candidate for biodiesel feedstock. In a preliminary study, the effects of four general microalgal growth factors, including temperature, pH, light intensity, and concentration of nitrogen source (KNO3), on the microalgal photoautotrophic growth were evaluated. With the exception of light intensity, the growth factors needed to be optimized for the microalgal biomass production. Optimization was done using response surface methodology. The optimal conditions for biomass production were pH 6.54, 27.66℃, and 0.52 g/l KNO3. The biomass production at the optimal conditions was 1.55 g/l, which correlated well with the predicted value of 1.5 g/l. The total lipid and fatty acid methyl ester contents of the cells grown at the optimal conditions were 49% and 21.2% of cell dry weight, respectively. To increase the lipid content of the biomass, microalgae were challenged by nitrogen starvation. Enhancement of total lipid and fatty acid content up to 52.02% and 49%, respectively, were observed. Lipid analysis of the nitrogen-starved cells revealed that C16 and C18 species accounted for 95.9% of the total fatty acids. Among them, palmitic acid (46.17%) and oleic acid (39.43%) dominantly constituted the algal fatty acids. These results suggest Desmodesmus sp. KAERI-NJ5 as a promising feedstock for biodiesel production.