Abstract: In this study,hind hoof meat was used to produce bundle hoof with Alcalase as curing agent and the process was optimized by response surface method.Enzymatic hydrolysis of Alcalase on bundle hoof protein was analyzed by the method of SDS-PAGE electrophoresis and free amino acids released. The result showed that Alcalase could promote proteolysis and improve the sensory quality of bundle hoof with pickled temperature at4~16 ℃. Bundle hoof protein could be excessive hydrolysis and had bad sensory quality with the Alcalase dosage higher than 1.6 U / g. The result of bundle hoof processing optimized by test of response surface was Alcalase dosage was 1.32 U / g and pickled temperature was 14.2 ℃ and pickled time was 21.3 d,then PI was 14.81% and sensory evaluation was 92.47. SDS-PAGE electrophoresis showed protein hands with higher than 120,30 ~ 40 ku and 50~70 ku were decomposition significantly,then Ala,Leu,Val,Tyr,Phe,Ile,Gly,His was significant increased and Pro,Glu( p < 0.05) was decreased significantly. It was could be conclude that Alcalase could significantly promote the decomposition of bundle hoof protein and improve the sensory quality of the product.So the Alcalase could be applied to bundle hoof production.