In this work, the enzymatic hydrolysis of bovine bone collagen was studied by pre-purified bone-specific collagenase (BSC). As evaluated in terms of degree of hydrolysis. The enzymatic hydrolysis process was studied through single factor and quadratic general rotary unitized design experiments. The results showed that the optimum hydrolysis conditions were as follows: reaction temperature 46 ℃, the amount of bovine bone collagen 5.14 g/100 mL, the amount of BSC enzyme 0.42 g/100 mL, reaction time 6 h, pH 6.5. Under these conditions, the degree of hydrolysis value was 34.98%. The structural properties of bovine bone collagen and its hydrolysate were characterized by ultraviolet, fluorescence, fourier transform infrared spectroscopy, and differential scanning calorimetry. The UV analysis showed that the BSC degradation enzyme damage the bone collagen three helix structure, a large number of free amino acid; FS analysis showed that the collagen molecular surface C═O, CONH₂ and COOH gradually increased, collagen peptide chromophore distribution also changes; FT-IR analysis showed that the degrada-tion of collagen from bovine bone collagen peptide after the the peptide chain of —NH⁺₂— repulsion effect gradually weakened, mainly in the beta angle; SEM showed that the enzyme damaged the surface of bovine collagen molecular structure, and made it loose.