Fruit and vegetable juice were used to improve the tenderness and prolong the shelf life of beef. Six twenty-month old Jin Jiang yellow cattles were selected, and their longissimus were taken as experimental materials after slaughter. the single factor experiment was carried out with the added tomato juice at 4%, 8%, 12%, 16% and 20%, respectively, ginger juice at 2%, 4%, 6%, 8% and 10%, respectively, kiwi fruit juice at 3%, 6%, 9%, 12% and 15%, respectively, papain juice at 5%, 10 %, 15%, 20% and 25%, respectively. Based on the single factor experiment, the pH, TVB-N (tomato and ginger processing group), tenderness (kiwifruit and papaya processing group) and sensory scores (storage at 3 d) were used as indicators to get the optimal conditions by response surface methodology. According to the Box-Behnken response surface optimization test, the optimum ratio of the composite vegetable and fruit modifying agent was 11.10% tomato, 6.53% ginger, 6.70% kiwifruit and 10.15% papaya juice, and the sensory score of beef was 95.10. The addition of tomato, ginger, kiwi and papaya significantly affected the sensory score of beef. Fruit and vegetable juice composite modifier optimized by response surface methodology was effective. It could be used as a kind of natural food additive to improve beef quality and prolong the shelf life.