Single cell oils (SCO) are a promising source of oils that could be exploited in different industrial areas. SCO for biodiesel production circumvents the controversy food vs. fuel, does not require large land areas for culture, and is independent of climate and seasonal variations, among other advantages in comparison to vegetable oils. In this study, a red yeast isolated from a mountain water source, identified as Rhodotorula glutinis T13, showed high potential for lipid production (40% w/w) with suitable growth parameters, yields, and fatty acids profile. Yeast lipids showed a high content of unsaturated fatty acids (56.44%; C18:1, C18:2), and the fuel properties (cetane number, iodine value, density, kinematic viscosity, etc.) of yeast oil analysed were in good agreement with international biodiesel standards. The results show that R. glutinis T13 can be used in the future as a promising microorganism for the commercial production of biodiesel.