Global energy is undergoing a transformation driven by the imperative to mitigate climate impact, enhance energy security, and promote sustainable development. Biofuel production has emerged as a promising alternative to fossil fuels, offering renewable and environmentally sustainable sources of energy. Biofuels are derived from organic materials such as crops, agricultural residues, forestry waste, and microorganisms, which undergo conversion processes to produce liquid or gaseous fuels suitable for transportation, heating, and electricity generation. Th is study aims to maximi ze the production of biomass, protein, lipid and carbohydrate in a newly isolated micr oalga, Tetradesmus obliquus RESP2 cultivated using a combination of municipal waste water and Bold's Basal Medium (BBM). The experimental design involved at varying the composition of growth medium to determine the most optimal conditions for algal growth and biochemical accumulation. After thorough experimentation and analysis, the results revealed that the optimal combination of waste water and BBM medium (75%:25%) yielded a maximum concentration of biomass 16.2 g/L, lipid 5.53 g/L, carbohydrate 1.48 g/L and protein 4.48 g/L. These findings demonstrate the efficacy of utilizing municipal wastewater as a nutrient source in combination with BBM medium for the cultivation of T. obliquus The optimized conditions identified in this study hold promise for enhancing the efficiency and sustainability of microalgae based biofuel production processes.