One of the major issues with a sustainable environment is oil spills. Membrane technologies are increasingly being used to purify water and treat wastewater. A hydrophilic and extremely porous substance, bacterial cellulose (BC) has promise for oil-water separation from contaminants. In our investigation, Komagataeibacter intermedius MO generated BC. Based on graphene oxide (GO) and bacterial cellulose (BC), an oil-water separation sheet has been created. The modified Hummer method was used to prepare GO, which was then added to BC sheets. The prepared sheets have been investigated by FTIR, SEM, and XRD. Scanning electron microscope analysis showed that the BC surface appears as a homogenous surface with small and middle fibers, while these fibers disappeared due to the presence of oil on the surface of BC/GO sheet after oil separation. Results revealed that in just five seconds, the generated sheets demonstrated remarkable efficiency in separating water from oil. Therefore, this study offers a promising technique for the application of BC-GO membranes for water and oil separation.