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<i>Chlorella vulgaris</i> and <i>Scenedesmus</i> sp. are commonly used in wastewater treatment due to their fast growth rates and ability to tolerate a range of environmental conditions. This study explored the cultivation of <i>Chlorella vulgaris</i> and <i>Scenedesmus</i> sp. using wastewater from the food industry, particularly from Italian basil pesto production tanks. The experiment involved different carbon dioxide concentrations and light conditions with a dilution rate of basil pesto wastewater at 1:2. Both microalgae strains were able to grow on pesto wastewater, and biomass characterization highlighted the influence of CO₂ supply and light irradiation. The highest lipid storage was 79.3 ± 11.4 mg g_{dry biomass}⁻¹ and 75.5 ± 13.3 mg g_{dry biomass}⁻¹ for <i>C. vulgaris</i> and <i>S. obliquus</i> under red light (5% CO₂ supply) and white light (0.04% CO₂ supply), respectively. Protein storage was detected at 20.3 ± 1.0% and 24.8 ± 1.3% in <i>C. vulgaris</i> and <i>S. obliquus</i> biomasses under white light with a 5% CO₂ and 0.04% CO₂ supply, respectively. The removal of P, N, chemical oxygen demand, and biological oxygen demand resulted in 80–100%, 75–100%, 26–35%, and 0–20%, respectively.