Abstract: In the enzymatic unhairing process, the risk of loosen and grain damage is mainly due to the excessive hydrolysis of collagen fibers by enzymes. In this experiment, compounding inhibitors were applied to reduce the excessive hydrolysis of collagen fibers during enzymatic unhairing process and to reduce the grain damage. The effect of compounding inhibitors on the enzymatic hydrolysis activity of AS1.398 were explored, and the compounding inhibitors based on Cu(II) ions were screened out and applied on bovine hide unhairing process. The results showed that the addition of 5 mmol/L of Cu(II) ions and 20 mmol/L of sodium succinate compounding inhibitor could maintain the enzymatic unhairing rate while inhibit the hydrolysis of collagen fibers. This study provides a potential novel strategy for industrial application of enzymatic unhairing process with safe and efficiency.