To study and optimize the fermentation parameters for expressing human-like collagenⅡduring E. coli high-density fermentation. The effects of pH, temperature, dissolved oxygen and induction instant on the cell growth and human-like collagenⅡproduction were investigated to optimize the fermentation conditions. The results demonstrated that the following conditions were beneficial for cell growth and foreign gene expression, controlling pH in phase induction at 6.8 and initial pH at 6.5, maintaining fermentation temperature and dissolved oxygen concentration was controlled at 34°C and 20% respectively, and implementing induction at the later logarithmic growth phase. Under the optimized condition, the cell density and human-like collagenⅡyield could reach 88.4 g/L and 14.2 g/L, respectively.