Microalgae have a high growth rate, high CO₂ absorption capacity, and high content of chlorophyll, proteins, vitamins, mineral salts, carbohydrates, antioxidant substances, and fatty acids. In recent years, Chlorella vulgaris has been widely used as a feedstock for producing third-generation biofuels, such as bioethanol. Thus, this work aims to develop a strategy to increase the production scale of the microalgae Chlorella vulgaris grown in distilled reused water, supplemented with a modified BG-11 medium, to use biomass in the production of bioethanol. The total cultivation of 72 L presented a concentration of 0.415 g·L⁻¹·d⁻¹, with 61.32 g of final biomass. To improve carbohydrate extraction, the biomass was pre-treated with sulfuric acid at different concentrations (1.5% and 3% v/v). The hydrolyzed solution was supplemented with YPD (yeast extract peptone dextrose) medium and inoculated with Saccharomyces cerevisae yeast, initiating fermentation. In each sample, the Brix degree, cell concentration, reducing sugar concentration, and alcohol content were analyzed. The sample pre-treated with sulfuric acid 1.5% v/v was the one that presented the best result, with alcohol content after distillation of 68 °GL (Gay-Lussac). It appears that the cultivation of the microalgae Chlorella vulgaris in scale-up, with reused water, has high potential in the production of third-generation biofuel.

Microalgae harvesting by coagulation can use coagulant agents such as alum, synthetic polymers or biocoagulants. Biocoagulants have attracted the attention of researchers because they are natural, biodegradable, and promote high microalgal harvesting efficiencies. This study aims to optimize the harvesting of Chlorella vulgaris based on the dosage of the Opuntia cochenillifera extract and the choice of eluent for biopolymer extraction. The outdoor cultivation of C. vulgaris achieved a specific growth rate of 0.455 d⁻¹ and a maximum biomass concentration of 1.28 g_DW L⁻¹. In order to harvest the microalgal biomass, the polymer present in the mucilage of O. cochenillifera was extracted using NaOH and HCl. Coagulation and sedimentation assays were performed with different coagulant dosages: 3.5, 5.9, and 8.2 g L⁻¹. The maximum harvesting efficiencies using the acid and alkaline extract coagulant solutions were 80.8% and 99.5%, respectively, with a dosage of 3.5 g L⁻¹. According to the results, the C. vulgaris biomass can be harvested with the mucilage from O. cochenillifera in acid and alkaline eluents. The application of this biocoagulant constitutes a sustainable solution for microalgal harvesting.