In recent years, active biomolecules such as chitosan and its derivatives are undergoing a significant and very fast development in food application area. Due to recent outbreaks of contaminations associated with food products, there have been growing concerns regarding the negative environmental impact of packaging materials of antimicrobial biofilms, which have been studied. Chitosan has a great potential for a wide range of applications due to its biodegradability, biocompatibility, antimicrobial activity, nontoxicity and versatile chemical and physical properties. It can be formed into fibers, films, gels, sponges, beads or nanoparticles. Chitosan films have been used as a packaging material for the quality preservation of a variety of foods. Chitosan has high antimicrobial activities against a wide variety of pathogenic and spoilage microorganisms, including fungi, and Gram-positive and Gram-negative bacteria. A tremendous effort has been made over the past decade to develop and test films with antimicrobial properties to improve food safety and shelf-life. This review highlights the preparation, mechanism, antimicrobial activity, optimization of biocide properties of chitosan films and applications including biocatalysts for the improvement of quality and shelf-life of foods.