Development of biodegradable and biocompatible films based on the proteinpolymer with strong antibacterial activities is gradually obtained extensive concern in the world. The aim of this study was to evaluate the antioxidant and antibacterial properties of gelatin nano films merged with different concentrations of Zataria multiflora essential oil. Gelatin films were prepared from gelatin solutions (10% w/v) containing Zataria multiflora essential oil [ZMO] (2, 4, 6 and 8% w/w), glycerol (25% w/w) as plasticizer, and glutaraldehyde (2% w/w) as cross-linker. The mechanical, water solubility, water swelling, water vapor permeability, antioxidant and antibacterial properties of the films were measured according to the American Society for Testing and Materials. Gelatin films exhibited good tensile strength and elongation at break, water solubility, swelling, and water vapor permeability. Incorporation of ZMO into the gelatin films caused a significant decrease in tensile strength and swelling, and a significant increase in elongation at break, water solubility, water vapor permeability and whiteness of the films. Gelatin films exhibited low antioxidant activity while gelatin films incorporated with ZMO exhibited excellent antioxidant properties. The films incorporated with ZMO also exhibited excellent antibacterial properties against both Gram-positive and Gram-negative bacteria. Our results suggest that the gelatin/ZMO films could be used as a very attractive alternative to traditional materials for different biomedical applications.