The challenge for food industry is developing gentle processes concept, which will prevent food spoilage and leave a food natural, minimally processed, fresh-like and safe. A new technique of food preservation could be usage of combined methods of green process, such as cold gas plasma with bioactive substances, and protective coatings. The aim of this study was to determine the antibacterial activity of chitosan films incorporated with lysozyme exposed to helium plasma treatment as well as evaluate their physical properties. The edible films have been prepared basing on low molecular weight chitosan by casting from lactic acid solution with water solution of lysozyme in three various concentrations (0, 0.5 and 1 %). Dried films were then modified by exposition on cold helium plasma treatment for 0, 5 and 10 min. Obtained films were tested against growth of Listeria monocytogenes, Yersinia enterocolitica and Pseudomonas fluorescens. In order to characterize chitosan-based films, their mechanical properties, theromogravimetric analysis (TGA), dynamic mechanical thermal analysis (DMTA), contact angle measurement, water vapour permeability (WVP) and scanning electron microscopy (SEM) were evaluated. The films with 1 % lysozyme incorporation enhanced the inhibition efficiency of chitosanbased films against gram-positive (L. monocytogenes) and gram-negative (P. fluorescens) bacteria, where reduction zones were 42.5 and 69.8 mm, respectively. Besides that, hydrolytic changes of chitosan chain caused by lysozyme activity were confirmed by TGA and DMTA. Contact angles and WVP of tested films were not significantly affected by helium plasma exposition, nor lysozyme dosage. Hydrophilic natures of chitosan-based films were confirmed by both tests.Microscopy image of cross-section structure was smooth and continuous due to lysozyme addition in film composition. Application of chitosan films incorporated with lysozyme and low-pressure plasma treatment could be used as innovative preservation method in a wide range of food products.