Abstract:The tenderization of pre-conditioned hot pot beef using a mixture of papain and neutral protease was optimized by one-factor-at-a-time approach and response surface methodology. Mixing ratio between the two enzymes, enzyme dosage and treatment time were considered as independent variables, and water-holding capacity and hardness of beef as responses. It was found that the optimal process conditions were determined as follows: mixing ratio between papain and neutral protease 1:1.19, enzyme dosage 0.085%, and treatment time 3.1 h. Compared with the conventional product, the waterholding capacity of enzymatically tenderized beef increased by 27%, the myofibrillar fragmentation index (MFI) increased by 45%, the solubility of collagen increased by 29%, the shear force decreased by 29%, the hardness dropped by 69%, and the tenderness was significantly improved.