The development of food packaging films from nanocellulose has received attention from researchers. In this study, the chemo-mechanical method has been used to obtain nanocellulose from sugarcane bagasse. The shape and size of the nanocellulose were examined by field emission scanning electron microscopy (FE-SEM) using ImageJ software, and the result was found to be in good agreement with that of dynamic light scattering (DLS) analysis.Various characterizations such as X-ray diffraction (XRD), Fourier transform infrared (FTIR), thermo-gravimetric analysis (TGA), and ultraviolet visible (UV-Vis) spectrometer for the analysis of crystallinity, chemical structure, thermal stability, and optical properties were performed for fibers and nanocelluloses. Moreover, antibacterial evaluation was also carried out using the disc diffusion method with Bacillus and Escherichia coli bacteria. The removal of noncellulosic elements from fibers after treatment and grinding was revealed by FTIR analysis. Furthermore, nanocellulose exhibited not only higher thermal stability and crystallinity but also acceptable antibacterial properties. Based on the results of this study, polymeric films and composites of the presented nanocellulose might be utilized in food packaging, plastic packaging, and container fabrication.