Bananas are one of the most important crops worldwide. However, a large amount of residual lignocellulosic biomass is generated during its production and is currently undervalued. These residues have the potential to be used as feedstock in bio-based processes with a biorefinery approach. This work is based on the valorization of banana leaf and has the following objectives (i) to determine the effect of certain physical and environmental factors on the concentration of glucose present in banana leaf extract (BLE), using a statistical regression model; (ii) to obtain Bacterial Nanocellulose (BNC), using BLE (70% v/v) and kombucha tea as fermentation medium. In addition, the physicochemical properties of BNC were evaluated by X-ray diffraction (XRD), Fourier transform infrared (FTIR), and thermogravimetric analysis (TGA). The results indicate that storage time, location, leaf color, and petiole type are factors related to BLE concentration, which is reduced by approximately 28.82% and 64.32% during storage times of five days. Regarding BNC biosynthesis, the results indicate that the highest yield, 0.031 g/g, was obtained at 21 days. Furthermore, it was determined that the highest production rate was 0.11 gL−1h−1 at 11 days of fermentation. By FTIR, it was determined that the purification step with NaOH (3M) should be carried out for approximately two hours. This research supports the development of a circular bioeconomy around the banana value chain, as it presents a way of bioprocessing residual biomass that can be used to produce bioproducts.