An effective antibacterial nanofiber film was prepared through the incorporation of epsilon-polylysine (epsilon-PL) into gelatin/chitosan-based polymers. All nanofiber films had uniformly disordered fibrous structure with good diameter distribution. The weight ratio of the gelatin/chitosan/epsilon-PL (G/C/P) influenced the solution property and nanofiber morphology. The addition of epsilon-PL can decrease the viscosity and increase the conductivity of solutions, which lead to a decrease in the diameter of nanofibers. The three polymers of gelatin, chitosan, and epsilon-PL were interacted by hydrogen bonding, and the crystallinity of nanofiber films was decreased by the electrospinning process. The addition of epsilon-PL can enhance the thermal stability, and decrease the water vapor permeability and oxygen permeability of the films, and epsilon-PL did not easily release from the nanofiber films. The G/C/P (6:1:0.125) nanofiber film was more effective to control six foodborne pathogens than the G/C nanofiber films by destroying the bacterial cell membranes. The result indicated that the gelatin/chitosan/epsilon-PL nanofiber films can be used as a food-packaging material to reduce the risk of foodborne pathogens. (C) 2020 Elsevier B.V. All rights reserved.