Simultaneous saccharification and fermentation (SSF) is the most commonly practiced operation in lignocellulose bioconversion to avoid the sugar product inhibition to cellulase enzymes. In this study, for the first time SSF was tested on microbial lipid fermentation using the diluted acid pretreated and biodetoxified corn stover. The results show that SSF was effective than the separate hydrolysis and fermentation (SHF) on lipid accumulation of Trichosporon cutaneum CX1 cells in both the small scale (5L) and the enlarged scale (50L) bioreactors. The solutions for the oxygen transfer and the lipid extraction in SSF practically worked well. The process parameters were optimized and the lipid yield obtained were 3.03g/L in the 5L, and 3.23g/L in the 50L, respectively. The result also shows that the cellulase enzyme could be partially recycled in the SSF. The study provided a practical and efficient way for microbial lipid production from lignocellulose material.