This paper presents the results of research on the preparation and properties of GO/BC nanocomposite from bacterial cellulose (BC) modified with graphene oxide (GO) using the in situ method. Two bacterial strains were used for the biosynthesis of the BC: Komagataeibacter intermedius LMG 18909 and Komagataeibacter sucrofermentans LMG 18788. A simple biosynthesis method was developed, where GO water dispersion was added to reinforced acetic acid-ethanol (RAE) medium at concentrations of 10 ppm, 25 ppm, and 50 ppm at 24 h and 48 h intervals. As a result, a GO/BC nanocomposite membrane was obtained, characterized by tensile strength greater by 150% as compared with the pure BC (̴ 50 MPa) and lower volume resistivity of ~4 ∙ 109 Ω × cm. Moreover, GO addition increases membrane thickness up to ~10% and affects higher mass production, especially with low GO concentration. All of this may indicate the possibility of using GO/BC membranes in fuel cell applications.