We evaluated the optimal conditions for extraction of pepsin-soluble collagen from the skin of Chinese giant salamander () using a Box-behnken design experiment. The concentration of pepsin, solvent-to-solid ratio, and enzyme extraction time were the test factors and the extraction rate of collagen was the target parameter. We used response surface methodology to describe the influence of the three factors on the extraction rate of collagen and developed aquadratic polynomial regression model to optimize the extraction conditions. The order of influence of the factors was: solvent-to-solid ratio>enzyme extraction time>pepsin amount. The interaction between the solvent-to-solid ratio and enzyme extraction time had a significant effect on the extraction rate of collagen, while other interactions had no significant effect. The optimal conditions predicted by the regression model for extraction were slightly adjusted to meet actual production. The specific parameters were: pepsin concentration was 16.5%, solvent-to-solid ratio was 15 mL/g, and enzyme extraction time was 29 h. Under these conditions the extraction rate was 66.99%, which was similar to the theoretical value (67.83%) predicted by the regression model.