•Highest lipid was accumulated when mixed VFAs were used.•The lipid produced from mixed VFAs was higher than that from the sole acetic acid.•Both propionic and butyric acids can improve the odd-numbered fatty acid content.•The highest odd-numbered FAs yield was achieved when the VFA ratio was 5:15:10. The oleaginous yeast Cryptococcus curvatus ATCC 20509 can use 5–40g/L of acetic, propionic, or butyric acid as sole carbon source to produce lipids. High concentrations (30g/L) of mixed volatile fatty acids (VFAs) were used to cultivate C. curvatus to explore the effects of different ratios of mixed VFAs on lipid production and composition. When mixed VFAs (VFA ratio was 15:5:10) were used as carbon sources, the highest cell mass and lipid concentration were 8.68g/L and 4.93g/L, respectively, which were significantly higher than those when 30g/L of acetic acid was used as sole carbon source. The highest content and yield of odd-numbered fatty acids were 45.1% (VFA ratio was 0:15:15) and 1.62g/L (VFA ratio was 5:15:10), respectively. These results indicate that adjusting the composition ratios of mixed VFAs effectively improves microbial lipid synthesis and the yield of odd-numbered fatty acids.