The environmental problems caused by the disposal of synthetic polymers in the environment are of interest in the study of the development of biodegradable films that can act as packaging or as components of the same. Among the natural biopolymers most used to obtain films are proteins from renewable sources, capable of forming continuous and cohesive matrix. Among the various renewable sources, fish stands out. The aim of this research was to extract myofibrillar protein (MP) from byproducts of the yellow hake processing (Cynoscion acoupa) to evaluate the functional properties of the biodegradable film with different concentrations of glycerol. The MP was extracted from the yellow hake, lyophilized and later elaborated the biodegradable film in the concentrations of 5, 10, 15, 30 and 40% of glycerol. Water vapor permeability (WVP-g.mm/m2d.KPa), tensile strength (RT-MPa) and elongation (E%) of the films were performed. The results were treated statistically (ANOVA; Tukey's test). WVP presented the results (2.36±0.16b, 2.66±0.194bc, 2.78±0.14bc, 3.33±0.30ac, 3.37±0.17ac), RT (28,06±0,48a; 26,28±2,50ac; 20,76±4,73cd; 15,84±0,23bd; 10,32±0,41b), and E (108±0,41bcd; 110,47±1,27bcd; 111,57±1,12bc; 112,35±0,20ac; 113,96±0,67a) at the concentrations of 5, 10, 15, 30 and 40% glycerol, respectively. Under these conditions, the films prepared with higher concentrations of plasticizers showed differences (p≤0.05) in comparison to the film with a lower concentration, resulting in higher WVP. RT there was a significant influence (p≤0.05) for all the results, presenting a higher RT in the lowest concentrations of plasticizers. While for the E, a difference (p≤0.05) was obtained at the extremes, with higher percentage of E in the highest concentration of plasticizer. The properties of the biodegradable films were affected by the composition of the film-forming solution, in particular glycerol. The barrier and mechanical properties of the films depend on the characteristics of the biopolymer and on the interaction between the polymer and the permeant, associated with polarity.