This study investigated the effects of different combinations of sous-vide cooking temperature (11, 14 and 17 h) and time (57, 60 and 63 ℃ ) on the eating quality of beef Semimembranous muscle in terms of pH value, internal color of cooked meat, cooking loss, tenderness, and sensory scores. The results showed that as cooking time and cooking temperature increased, the pH value, shear force, cooking loss, L* value, and H* value of bovine Semimembranous muscle all showed an increasing trend, while the a*, b*, C*, and overall sensory score significantly decreased. The cooking loss increased from 28.98% to 33.06% over 11 to 17 hours, and from 27.2% to 36.5% over 57 to 63 ℃ ; the shear force increased from 40.14 to 61.08 N over 57 to 63 ℃ , indicating that as the cooking strength increased, both the water-holding capacity and tenderness decreased. At 57 ℃ and 11 hours, cooked beef had the lowest shear force of 42.58 N, which meets the demand of Chinese consumers for “tender meat”, and the highest scores for overall acceptability and internal color. Therefore, cooking at 57 ℃ for 11 hours can effectively improve the tenderness and water-holding capacity of beef Semimembranous muscles while avoiding some problems such as the reddish color and bland flavor of cooked meat and imparting it with better sensory quality and higher yield.