Bilayer films propose to combine the characteristics of different individual polymer sources in a single structure, aiming to improve the properties of the films obtained. The objective of this study was to develop and characterize bilayer films prepared from individual films of zein, wheat gluten, and protein isolate from hake (Cynoscion guatacupa), and evaluate the biodegradability of these polymers. Through the casting technique, individual films and bilayers were prepared. The hake proteins isolate films, as well as the bilayers hake protein isolate/zein and hake protein isolate/wheat gluten (BI PI/WG) showed higher tensile strength, while wheat gluten films presented greater elongation at rupture and lower solubility. In addition to having good tensile strength, BI PI/WG presented the best elongation. All films and bilayers reached melting and glass transition temperatures above 100°C. Electromicrographs of the films presented an irregular surface with pores and grooves, being less pronounced in BI PI/WG. In the bilayers, it was not necessary to use any type of adhesive to form a single structure. Films and bilayers showed total biodegradability in 40 days, with the exception of the individual zein film. BI PI/WG proved to be the most suitable for the development of food packaging materials. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018, 135, 46721.