Packaging is first protective against spoilage of food after producing. One of the ways to improve the protective properties of packaging material is to utilize antimicrobial components as the major component. Nowadays, because of increasing of population, used material for packaging of food have high volume of waste. Most of these components are petroleum derivatives which are time- consuming and expensive in recycling that lack of them leads to environmental consequents. This agent reinforces attempts to use biodegradable components which nano-technology with new ideas paves the ways to cover imperfects of packaging industry. In this study, biodegradable polymer chitosan and ethylene vinyl alcohol (EVOH) for film matrix and ZnO as filing material was used. Agent glycerol was used as softening material. The film was produced by dissolving in solvent and oven was utilized for evaporating.in addition, chitosan and nano zinc oxide have antimicrobial properties. in this study, five level of 100: 0, 75: 25, 50: 50, 25: 75 and 0: 100 of chitosan / ethylene vinyl alcohol and three levels of 0, 1% and 2% nano zinc oxide and three levels of 30%, 40% and 50% glycerol was used. After producing film, mechanical and physical and antimicrobial activity properties were evaluated. By adding EVOH to chitosan water vapor and oxygen permeability significantly decreased. Also traarency and elongation of film was improved. EVOH reduce necessity force to rupture of tear film and time was increased (increase flexibility) increasing in amount of glycerol had same action. Adding chitosan to EVOH leads to enhance resistance against water and moisture.by adding nano particle amount of water vapor and oxygen permeability and was decreased which had best case in 1%. Furthermore, nanoparticle improved mechanical properties.adding glycerol leds to increase of permeability of films and was increased through increase of concentration. Glycerol reduced tensile strength. Produced and possible bonds caused from interaction of these components were examined by FT-IR. The physical and mechanical variations in film were explained by these components. Cross-section of film was examined by FESEM microscope befor and after mixing. Also dispersion of nanoparticle was examined.antimicrobial propertie of these components were examined by Aspergillus niger mold and Escherichia coli bacterium which had significant inhibition versus mold and bacterium growth.chitosan because of amino groups have positive charge and zinc nanooxide have antimicrobial properties because of influence on microbe DNA . By increasing chitosan and zinc nano oxide, antimicrobial and mold properties was increased. Zinc nano oxide had high mold and antimicrobial properties.