Abstract: According to the nutritional needs of Bifidobacterium adolescentis,the medium for Bifidobacterium adolescentis were optimized by single factor experiment. The optimum medium prescription was confirmed as follows：2% sucrose,1% casein peptone, 1% soya peptone, 0.7% yeast extract, 0.3% xylooligosaccharides, 0.05% KH 2 PO 4 , 8% cabbage juice. The growth curve of B. adolescentis in the optimum medium was determined. The changes of pH and absorption value were determined. It was confirmed that the terminal time for the fermentation culture was 14-16 h hours and the number of active bacteria could reach 2.0 ×10 9 cfu/mL. The optimum freeze -drying protective agents were confirmed by single factor test and orthogonal test ： 20% trehalose,20% skimmed milk powder,1.0% glycerin,8% sucrose,5% gelatin.