Yellowfin tuna skin waste is a potential collagen source for gelatin through hydrolysis. Enzymatic hydrolysis is eco‐friendly but expensive, necessitating efficiency improvements. Ultrasound pre‐treatment enhances efficiency via cavitation effects. This study evaluated the effects of ultrasound amplitude, pre‐treatment duration, enzyme concentration, and hydrolysis duration on yield; determined optimal conditions for ultrasound‐assisted enzymatic extraction on yield, viscosity, and gel strength; and assessed the gelatin's physicochemical and functional properties. This research consists of three stages, factor screening using one variable at a time, optimization using response surface methodology with Box‐Behnken design, and gelatin characterization. The study included three replicates. Screening identified factor ranges for optimization: 5–25 min pre‐treatment, 1%–3% enzyme concentration, and 1–5 h hydrolysis. Optimization involved 15 experimental runs, focusing on yield, viscosity, and gel strength. Optimal conditions were 25 min pre‐treatment, 1% enzyme concentration, and 2.17 h hydrolysis, yielding 64.08%, 9.29 mPa·s viscosity, and 166.28 bloom gel strength. The optimized gelatin showed emulsifying properties with EAI 50.5 m2/g and ESI 23.80 min, water‐oil holding capacities of 192% and 406%, respectively, antioxidant activity of 60.16%, and molecular weight range of 17–160 kDa. The amino acid composition was mainly glycine (29.26%) and proline (18.26%), with 76.95% digestibility. This innovative, environmentally friendly extraction method improved gelatin extraction efficiency from yellowfin tuna skin, supporting a sustainable industry.