Abstract: In the present study,the single factor experiment and orthogonal experiment were used to optimize the cultivation conditions for feed yeast strain YS-01 with sugarcane molasses. The results revealed that the feed yeast strain showed good growth in the treated molasses; the optimal culture conditions were： molasses concentration was 10^0 Bx, inoculation amount was 8.0%,fermentation duration was 12 h; the optimum concentrations of various nutrients were：（NH4）2SO4 was 0.10%, CO（NH2）2 was 0.10%, KH2PO4 was 0.16%, MgSO4·7H2O was 0.05%. Under this condition, the OD value reached 14.670 and the cell density reached 2.55×10^-8CFU/m L.