Lipid homeostasis is well-known in oleaginous yeasts, but there are few non-oleaginous yeast models apart from Saccharomyces cerevisiae. We are proposing the non-oleaginous yeast Candida zeylanoides QU 33 as model. The aim of this study was to investigate the influence of the carbon/nitrogen ratio and the type of nitrogen source upon oil accumulation by this yeast grown on shake flask cultures. The maximum biomass was obtained in yeast extract (2.39 ± 0.19 g/l), followed by peptone (2.24 ± 0.05 g/l), while the highest content of microbial oil (0.35 ± 0.01 g/l) and the maximum lipid yield (15.63%) were achieved with peptone. Oleic acid was the predominant cellular fatty acid in all culture media (>32.23%), followed by linoleic (>15.79%) and palmitic acids (>13.47%). The highest lipid yield using glucose and peptone was obtained at the C/N ratio of 200:1.