Purpose: The objective of this work was to carry out the cultivation of the microalgae Chlorella sorokiniana with diluted vinasse, verifying its potential for remediation of nutrients and increase of its lipid fraction, to produce biodiesel according to the current regulation. Theoretical Framework: The quest to reduce fossil fuels, which are responsible for negative environmental impacts, has boosted the demand for biofuels in recent years. Biodiesel has been gaining prominence as a promising alternative to replace petroleum derivatives since it can be produced from various raw materials, such as microalgae. Design/Methodology/Approach: Different vinasse dilutions were studied for the cultivation of Chlorella sorokiniana CTT 7727, being 5, 10 and 15% (v v-1) and N:P:K (20:5:20) 1% (v v-1) medium as a comparative standard. Biodiesel was produced by direct transesterification and analyzed by gas chromatography coupled to a flame ionization detector, enabling the identification and quantification of the fatty acid profile. Findings: The results obtained indicated that vinasse 10% (v v-1) proved to be promising for application as a culture medium for this microalgae species, as it guaranteed a good cell growth rate, high biomass productivity and lipid yield. Considering the biodiesel produced, it fits the regulatory standards analyzed by the National Agency of Petroleum, Natural Gas and Biofuels (ANP). Research, Practical & Social Implications: The research demonstrates the feasibility of cultivating the microalga Chlorella sorokiniana with diluted vinasse, highlighting the 10% concentration as promising for optimizing cell growth and lipid yield. The results ensure compliance with ANP regulations for biodiesel production, indicating immediate and sustainable applicability on an industrial scale. Socially, it contributes to the pursuit of biofuels, reducing dependence on fossil sources and promoting more sustainable practices in the bioenergy industry. Originality/Value: The study's originality lies in the unique combination of Chlorella sorokiniana CTT 7727, diluted vinasse, and biodiesel production in accordance with ANP regulations. It stands out for identifying the optimal vinasse concentration (10%) for cultivation, presenting an economical and effective approach. The strategic value is in the use of an alternative raw material cultivated with an industrial residue, promoting the diversification of the energy matrix and mitigating environmental impacts associated with conventional fuels.