Oils from yeasts have emerged as a suitable alternative raw material to produce biodiesel, due to their similar composition to common raw materials such as vegetable oils. Additionally, they have the advantage of not competing with human or animal feed, and, furthermore, they do not compete for arable land. In this work, a carbon and energy balance was evaluated for Yarrowia lipolytica as a model yeast, using crude glycerol from biodiesel as the only carbon source, which improves biodiesel overall yield by 6%. The process presented a positive energy balance. Feasibility of yeast oil as biodiesel substrate was also evaluated by determination of the lipid fatty acid profile and cetane number. Moreover, a comparison of oil yields, in terms of land use, between vegetable, microalgae, and yeast oils is also presented. The results showed that Y. lipolytica oil yield is considerably higher than vegetable oils (767 times) and microalgae (36 times).