Cellulose nanofibers were isolated from banana peel using a combination of chemical and mechanical treatments with different number of passages through the high-pressure homogenizer (0, 3, 5, and 7 passages). New nanocomposites were then prepared from a mixed suspension of banana starch and cellulose nanofibers using the casting method and the effect of the addition of these nanofibers on the properties of the resulting nanocomposites was investigated. The cellulose nanofibers homogeneously dispersed in the starch matrix increased the glass transition temperature, due to the strong intermolecular interactions occurring between the starch and cellulose. The nanocomposites exhibited significantly increased the tensile strength, Young's modulus, water-resistance, opacity, and crystallinity as the number of passages through the homogenizer augmented. However, a more drastic mechanical treatment (seven passages) caused defects in nanofibers, deteriorating the nanocomposite properties. The most suitable mechanical treatment condition for the preparation of cellulose nanofibers and the corresponding nanocomposite was five passages through the high-pressure homogenizer. In general, the cellulose nanofibers improved the features of the starch-based material and are potentially applicable as reinforcing elements in a variety of polymer composites.