Abstract: Extracting process of astaxanthin from Phaffia rhodozyma was optimized from disruptive methods, extraction solvents and extracting conditions. Results showed that dimethylsulphoxide (DMSO) method was the best to disrupt cell walls and acetone was suitable solvent for extracting astaxanthin from this yeast. Results of factor design experiment indicated that factors such as disrupting temperature, disrupting time, quantity of acetone and extracting temperature had significant effects on the extracting rate, and disrupting temperature and disrupting time were the most significant for the extracting rate compared with the other factors. The optimal condition described as that DMSO addition was 25 mL/g(dry cells), temperature for breaking cell walls 75.6℃, time for disrupting cells 20min, acetone addition 77.4 mL/g(dry cell), extracting temperature 40℃ was obtained by the steepest ascent experiment and the central composite design. Under the optimized condition, the astaxanthin concentration of the extracting liquid was 3.145 μg/mL, which increased 27.02% comparing with that of the non-optimized condition.