Abstract: In this study, three kinds of biological preservatives（ε-polylysine, nisin, natamycin） were selected to extend the shelf life of cake, in order to improve the edible safety. The preservation effects of ε-polylysine,nisin and natamycin used alone on chiffon cake were investigated by single factor tests, and the optimal amounts of three preservatives used together were obtained by response surface test design, and the preservation effects were compared with that of potassium sorbate and sodium dehydroacetate commonly used in cake. Results showed that independently used natamycin and ε-polylysine could significantly reduce the total number of mold in cake,nisin and ε-polylysine could remarkably decrease the total number of colonies in cake when used alone. The optimal dosages of the three kinds of preservatives used together were determined as follows: ε-polylysine addition of0.106 g/kg, nisin spraying of 0.108 g/L, and natamycin spraying of 0.114 g/L. The combination could significantly extend the shelf life of cake from 2 days to 5 days at 37 ℃, and the combination showed superior fresh-keeping effects compared with potassium sorbate and sodium dehydroacetate. This study could provide a certain theoretical reference for the applications of biological preservatives in cake, and open a new way for the preservation of cake.