This study evaluated the biofuel production potential of two algal species, Chlorella pyrenoidosa and Scenedesmus abundans, under stress conditions induced by nutrient supplementation or starvation at varying light intensities. Central composite face-centered design response surface methodology (CCFD-RSM) was employed to optimize stress conditions by varying the sodium nitrate (NaNO3), potassium dihydrogen phosphate (KH2PO4), dipotassium hydrogen phosphate (K2HPO4), cultivation time, and light intensity. The study included both C. pyrenoidosa and S. abundans, which presented increased biomass yields when subjected to nutrient starvation. Under the optimized conditions, the dry biomass yield was 98.26mg/L for C. pyrenoidosa and 110mg/L for S. abundans. Lipid yields were approximately 22.47% for C. pyrenoidosa and 29.06% for S. abundans under these optimized growth conditions. The optimized parameters for maximum biomass and lipid production were identified as C. pyrenoidosa, and the optimized conditions required 0.805g/L NaNO3, 0.052g/L K2HPO4, 0.099g/L KH2PO4, 17days of culture, and 5168.39lx of light intensity. For S. abundans, the optimal conditions were 1.065g/L NaNO3, 0.071g/L K2HPO4, 0.058g/L KH2PO4, 22days of cultivation, and 2897lx of light intensity. Overall, both C. pyrenoidosa and S. abundans have emerged as promising candidates for sustainable biodiesel production, highlighting their potential under stress conditions induced by nutrient modulation and variable light intensities.