Interest in nanocellulose obtained from natural resources has grown, mainly due to the characteristics that these materials provide when incorporated in biodegradable films as an alternative for the improvement of the properties of nanocomposites. The main purpose of this work was to investigate the effect of the incorporation of nanocellulose obtained from different fibers (corncob, corn husk, coconut shell, and wheat bran) into the chitosan/glycerol films. The nanocellulose were obtained through acid hydrolysis. The properties of the different nanobiocomposites were comparatively evaluated, including their barrier and mechanical properties. The nanocrystals obtained for coconut shell (CS), corn husk (CH), and corncob (CC) presented a length / diameter ratio of 40.18, 40.86, and 32.19, respectively. Wheat bran (WB) was not considered an interesting source of nanocrystals, which may be justified due to the low percentage of cellulose. Significant differences were observed in the properties of the films studied. The water activity varied from 0.601 (WB Film) to 0.658 (CH Film) and the moisture content from 15.13 (CS Film) to 20.86 (WB Film). The highest values for tensile strength were presented for CC (11.43 MPa) and CS (11.38 MPa) films, and this propriety was significantly increased by nanocellulose addition. The results showed that the source of the nanocrystal determined the properties of the chitosan/glycerol films.