The potential use of banana peel to produce cellulose nanofibers was investigated. A process conditions study to obtain nanofibers by enzymatic hydrolysis was conducted using a 24-1 fractional factorial design. Alkaline treatment was a necessary step to remove the amorphous fraction present in the fiber. Optimization of the enzymatic hydrolysis process conditions enabled production of typical cellulose nanofibers. Banana peel nanoparticles have potential application as reinforcing elements in composites.