There is an increasing interest toward biodegradable active packaging because of consumer demand and environmental concerns. Despite this interest, poor thermal, mechanical, and water barrier properties of biodegradable polymers such as gelatin limit their application in food packaging. In this study, to prevail these limitations, the gelatin-based nanocomposite containing chitosan nanofiber (CHNF) and ZnO nanoparticles (ZnONPs) were fabricated and characterized by FTIR, SEM, and DSC analyses. The results showed the appropriate interactions between gelatin matrix, CHNF and ZnONPs due to their good compatibility. Additionally, the nanocomposite showed high mechanical and water barrier properties due to its high dense and less permeable structure. The incorporation of CHNF compensated the negative effect of ZnONPs on the color properties of gelatin film. In addition, the synergistic effect between CHNF and ZnONPs improved the antibacterial activity of nanocomposite. In conclusion, the fabricated bio nanocomposite indicated considerable potential for food packaging.