The unconventional yeast Yarrowia lipolytica is known for its capacity to produce citric or isocitric acid from glycerol. In this study a reduction of production cost was achieved by using cheap crude glycerol and conducting the production at pH 3 to prevent bacterial contamination. In this study a Y. lipolytica strain overexpressing Gut1 and Gut2 was used. For the modified strain, crude glycerol proved to be an excellent substrate for production of citric/isocitric acids in aseptic conditions, as the final concentration of these compounds reached 75.9 +/- 1.8 g L-1 after 7 days of batch production. Interestingly, the concentration of isocitric acid was 42.5 +/- 2.4 g L-1, which is one of the highest concentrations of isocitric acid obtained from a waste substrate. In summary, these data show that organic acids can be efficiently produced by the yeast Y. lipolytica from crude glycerol without any prior purification in aseptic conditions.