This paper reports the production of lipids from non-hydrolyzed levoglucosan (LG) by oleaginous yeasts Rhodosporidium toruloides and Rhodotorula glutinis. Enzyme activity tests of LG kinases from both yeasts indicated that the phosphorylation pathway of LG to glucose-6-phosphate existed. The highest enzyme activity obtained for R. glutinis was 0.22 U/mg of protein. The highest cell mass and lipid production by R. glutinis were 6.8 and 2.7 g/L, respectively from pure LG, and 3.3 and 0.78 g/L from a pyrolytic LG aqueous phase detoxified by ethyl acetate extraction, rotary evaporation and activated carbon. This corresponded to a lipid yield of 13.5 wt.% for pure LG and only 3.9 wt.% for LG in pyrolysis oil. (C) 2013 Elsevier Ltd. All rights reserved.