Objective: In this study, a Box-Behnken design was employed in order to optimize the concentration of sodium alginate (SA), low acyl gellan (LAG) and glycerol (GLY) as a function of tensile strength (TS) and elongation at break (EB) of the biofilms. Methods: A texturometer Shimadzu model EZ-Test EZ-S was employed to evaluate TS and EB by using constant displacement control with a crosshead speed of 30 mm/min. Rafael E. González-Cuello et al. Results: The regression model developed herein indicates that the selected variables were statistically determinants (p < 0.05) for the studied mechanical properties. The optimal concentration to obtain high TS values (134.38 MPa) were 1.00% (w/v) of SA, 1.00% (w/v) of LAG and 8.00% (v/v) of GLY. However, low values of EB (15.95%) were also obtained using the same concentrations of AS, GBA and GLY. Conclusions: By increasing the concentration of SA and LAG result in highly resistant biofilms with low flexibility while increasing the glycerol concentration and reducing the concentration of biopolymers yielded the opposite effect. The Box-Behnken design is a useful statistical method for improving the mechanical properties studied depending on the final application of the biofilms.