The effects of the ultrasound-assisted enzymatic method on yield and dissolution properties of collagen extracted from tilapia skin were studied. A single factor test was used to investigate the influence of ultrasonic power, enzyme dosage, and ultrasonic time on collagen yield. The optimum extraction process was obtained by using the box-Behnken experimental design. The collagen yield, ultraviolet spectrum, and solubility characteristics of ultrasonic pepsin-soluble collagen and pepsin-soluble collagen were compared. The optional extraction conditions were revealed in the study as follows: ultrasonic power at 165 W, ultrasonic time for 26 min, and enzyme dosage at 1.67%. Under these conditions, the yield of UPSC (62.26%) was significantly higher than that of PSC (29.02%). The ultraviolet spectrum showed that both of them had absorption peaks at 230 nm instead of 280 nm, which was consistent with the characteristics of collagen. Under different pH conditions, the relative solubility of UPSC was higher than that of PSC. The results show that the ultrasound-assisted enzymatic method is an efficient, environmentally friendly, and simple method for extracting collogen from tilapia skin, which is suitable for industrial production. This study not only provides the best technology for extracting collagen from tilapia skin by ultrasonic-assisted enzymatic method but also provides the basis for its application in food processing with high yield and good relative solubility of collagen.