In order to improve mechanical properties and extend food preservation performance of nanofibers, electrospun gelatin-glycerin-e-Poly-lysine nanofiber has been engineered in this study. The optimized ratio of gelatin: glycerin: e-Poly-lysine was found to be 25: 6: 10 (w/v/v) after the evaluation of electrospinning solutions and nanofiber physical properties. The tensile strength of nanofibers reached 1.63 +/- 0.04 MPa with addition of glycerin. The results of FTIR, SEM and AFM confirmed that nanofibers have been fabricated successfully and obtained uniform diameter at 204 nm. TGA and DSC were also analyzed, which revealed the nanofibers have stronger thermal stabilities than pure components. At last, the nanofiber was utilized to preserve beef and exhibited outstanding antibacterial activity against Listeria monocytogenes during 10-days storage at 4 degrees C and 7-days storage at 12 degrees C, without impact on the surface color and sensory properties. Therefore, electrospun gelatinglycerin- e-Poly-lysine nanofiber could be a prospective packaging material in food industry.