AbstractThe production conditions were optimized, and the characteristics of biodegradable chitosan-gelatin films crosslinked with various microbial transglutaminases (transglutaminase 1, which contains maltodextrin, and transglutaminase 2, which contains maltodextrin, a milk protein) were studied. The film properties depended on both the cross-linking conditions and the type and ratio of components in the mixture. Gels and films prepared from chitosan and gelatin with transglutaminase 1 had very low characteristics as compared to films obtained with transglutaminase 2. Infrared spectroscopy, X-ray tomography, and scanning atomic force electron microscopy showed that transglutaminase 2 forms cross-links between both gelatin and chitosan amino groups, which improves the film characteristics. Films prepared from chitosan, gelatin, and glycerol crosslinked with transglutaminase 2 had the following characteristics: a strength of 15.8 MPa, a tensile strength of 220.0%, a thickness of 73.1 μm, and a density of 1.9 g/cm3. The biodegradability of the films depended on the composition and ranged from 20 to 30 days. The resulting films can serve as a matrix to obtain medical materials with antibacterial and wound-healing properties.