This chapter reports the most relevant aspects related to the isolation, production, and characterization of bacterial nanocellulose (BC). Furthermore, various used strains for high-yield production of BC were summarized. Some vital parameters (e.g., mode of culture, type of bioreactor, media, etc.) for high-yield production of BC were further exemplified. Moreover, various characterization techniques were reported to understand the physiochemical properties of BC. Specific focus has been given to morphological, structural, and mechanical characterization, to utilize these properties for further application. © 2017 Elsevier Ltd. All rights reserved.