The goal of this study is to evaluate physical and antimicrobial properties of biopolymers prepared from gluten (WG) and methyl cellulose (MC) with incorporation of activated lactoferrin (ALF), rosemary extract (RE) and natamycin (NA). The effectiveness of agents and antimicrobial films were investigated by agar disc diffusion method. Mechanical and water vapor permeability (WVP) of antimicrobial films were also examined. Film structure was investigated by scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR). Application of films was studied on Kashar cheese inoculated with target molds during 30 days storage at 10oC.ALF and RE did not show inhibitory activity against tested microorganisms.Minimum inhibitory concentration (MIC) of NA was found 750 ppm against Aspergillus niger and Penicillium roquefortii. Both films containing NA showed antifungal activity concentrations at and above 2 and 1 mg NA/10 g film solution (fs) against A. niger and P. roquefortii, respectively. RE in combination with NA reduced MIC of NA against A. niger to 1.5 mg NA/10g fs in both films. FTIR spectroscopy did not indicate any interaction between NA and both films. SEM observations showed that NA crystallizes at high concentrations in biopolymers. There was no significant change in WVP of films containing NA. Incorporation of NA to WG film did not cause major changes in its mechanical properties. Reduction in tensile strength of MC films was observed at high NA concentration. Application studies demonstrated the efficacy of WG and MC films including NA against A. niger and P.roquefortii inoculated on Kashar cheese.