Objective: The extraction process of collagen peptides from the skin of swordfish （Xiphias gladius） was optimized in order to efficiently obtain collagen peptides with hyaluronidase inhibition activity by single factor and response surface method. Methods: The response surface method was used to optimize the preparation of collagen peptide with hyaluronidase inhibition activity. In this paper, the enzyme-soluble collagen was extracted from swordfish skin by pepsin, and then the salting-out method was carried out to purify the collagen initially. The specie of collagen was identified by polyacrylamide gel chromatography with 7.5% separation gel. After the basic test carried out by single-factor method according to the degree of hydrolysis and hyaluronidase inhibition rate, the hyaluronidase inhibition rate was selected as the response value for the optimization of hydrolysis conditions of collagen peptide derived from the skin of swordfish via Box-Behnken response surface experimental method. Results: The results showed that the pepsin-soluble collagen was identified as the type I collagen. The optimized hydrolysis conditions for the type I collagen peptide with hyaluronidase inhibition activity were: 6 h of hydrolysis time, 8∶2 of complex enzyme ratio, 32 ℃ of hydrolysis temperature and 6% of enzyme mixture addition. The hyaluronidase inhibition rate of the type I collagen peptide under these conditions was 42.35%. According to the statistical analysis, the response surface model fitted well and there was no significant difference between the predicted and actual values of hyaluronidase inhibition rate （P>0.05）. Conclusion: Therefore, this hydrolysis process could be applied to the production of type I collagen peptide with hyaluronidase inhibition activity derived from the skin of swordfish.