Daqu distiller’s grains were co-fermented as raw materials by Trichoderma.koningii and Trichosporon cutaneum to produce microbial oils,which can provide raw materials for bio-diesel development.The single factor tests were used to investigate the effects of ratio of strains,inoculum size,culture temperature and culture time on the production of microbial oils.The best processing conditions were obtained by orthogonal test through measuring the content of microbial oils in product.The result shows that the microbial oils content of the co-fermented product under the ratio of Trichoderma.koningii to Trichosporon cutaneum of 1∶1,the inoculum size of strains of 11％ at 28 ℃ for 6 d is 7.15 g/L.It is shown that the production of microbial oils co-fermented by Trichoderma.koningii and Trichosporon cutaneum with Daqu distiller’s grains is possible.The research provides a new idea for the reuse of Daqu distiller’s grains and also provides a new way for the development of microbial oils.