The microalgae Choricystis minor var. minor stand out as a potential source of lipids. In this study, it was evaluated the cultivate, on a pilot scale, the microalgae freshwater Choricistis minor var. minor, and the evaluation of biodiesel production from wet biomass using the pressure heating route of wet biomass followed by esterification and subsequent transesterification, as well as how the quality of the produced biodiesel was evaluated. The process of centrifugation was adequate since it allows obtaining a concentrated biomass with a water content of approximately 76%. The biomass showed high total lipid content (22%) and adequate conversion of these lipids into biodiesel. The pilot scale cultivation in open tanks showed a volumetric productivity with a production potential of six tons of biodiesel per hectare per year. Biodiesel production via the biomass pressure heating route followed by subsequent transesterification produced a biodiesel that meets most quality parameters. Some parameters still need to be adjusted, such as sulfur and ester contents. The processes of cultivation, isolation and conversion to biodiesel proposed in this work are an adequate alternative to the production of biodiesel using microalgae, with some improvements in the biodiesel purification process.