Fresh beef was used to study the effects of Curdlan and alcalase on shear force, cooking loss and sensory quality of chilled prepared beef, then the process was optimized. The result showed that, compared the control, Curdlan could has significant effect on the shear force, cooking loss and sensory quality of chilled prepared beef (P<0.05), but no significant on protein and alcalase had a significant hydrolysis of myofibrillar protein. Different level of Curdlan and alcalase had a significant (P<0.05) impact on the shear force, cooking loss of chilled prepared beef and could improve the sensory quality significantly (P<0.05). At last, the best tender process was Curdlan 5.43 mg/g, Alcalase 4.29 U/g and curing time 73.4 h from response surface results by quadratic regression and through the result of verification test, and the shear force, cooking loss, sensory evaluation was (0.96±0.006) kg, (19.96±0.36)%, 92.81±0.75, respectively. The relative error is less than 5%, which compared with the theoretical value. Then, the processes were accurate and apply to chilled prepared beef.