The corn stalk was fermented by Cellulomonas flavigena to produce reducing sugar, then the production was re-fermented by Trichosporon cutaneum to produce microbial oils. The best process conditions were obtained by orthogonal test through measuring the content of reducing sugar and microbial oils in product. The results show that the reducing sugar content of the corn stalk fermented by Cellulomonas flavigena is 5.720mg/mL under the inoculum of Cellulomonas flavigena of 5 % at 31°C for 3 d . The microbial oils content of the corn stalk re-fermented by Trichosporon cutaneum is 1.27g per 100g fermentation products under the inoculum of Trichosporon cutaneum of 11 % at 28°C for 7d . There is a certain feasibility to produce microbial oil fermented by Cellulomonas flavigena and Trichosporon cutaneum with corn straws. The research provides a new way for utilization of corn stalk and also provides a new method for development of microbial oils.