The ability of the oleaginous yeast Lipomyces starkeyi to efficiently produce lipids when cultivated on saccharified sweet sorghum stalks juice was evaluated. Initially the production of lipids using synthetic media mimicking sweet sorghum stalks has been studied and optimized concerning the nitrogen source and the C:N ratio. Under optimum conditions (yeast extract as nitrogen source and C:N ratio of 190) the lipid production reached 5.81 g/L with a lipid content of 47.3% (w/w) from a mixture of sucrose, glucose and fructose, mimicking the sugar composition of sorghum. When cultivated on sweet sorghum stalks juice, it was observed that no external nitrogen addition was necessary which could result in substantial decrease of the initial C:N ratio. Moreover a distinct saccharification process prior to yeast cultivation improved the lipid production yield as it resulted in an increase of the C:N ratio. The highest lipid production, which was 6.40 g/L with a lipid content of 29.5% (w/w), was obtained when juice from saccharified sweet sorghum stalks at an initial sorghum content of 12% (w/w) was used as feedstock.