Abstract: The influence of different dehairing methods on the yield, amino acid composition, secondary structure, gel strength, emulsification, and foaming properties of cowhide gelatin was analyzed. Dehairing methods including hot scalding, enzyme treatment, high-pressure processing, and high-pressure-assisted enzymatic treatment were compared. The dehairing method significantly influenced the structural and functional properties of the gelatin. Compared with hot scalding, high-pressure-assisted enzymatic treatment markedly altered the amino acid composition and secondary structure of cowhide gelatin through the synergistic actions of the enzyme and high-pressure processing. This method resulted in a 65.98% increase in surface hydrophobicity and a 16.47 J/g increase in heat enthalpy, while reducing the thermal denaturation temperature by 4.48 ℃ and the relative crystallinity by 7.77%. The microstructure exhibited numerous fragmented and curled sheet-like structures. These structural changes effectively improved the functional properties of gelatin. Among the methods tested, high-pressure-assisted enzymatic treatment achieved the highest yield (24.06%) and gel strength (632.83 g), and significantly improved the emulsification and foaming properties (P<0.05). Thus, high-pressure-assisted enzymatic treatment represents a promising strategy to enhance gelatin yield and functional properties, laying a foundation for the production of high-quality gelatin with greater industrial value.