Production of bacterial cellulose (BC) using sugar cane molasses (MO) with/without the presence of lignosulfonate (MOL) as a sole carbon source in a Hestrin-Schramm medium (HS) was investigated. Six strains of Acetobacter xylinum [American Type Culture Collection 10245 and Institute of Fermentation in Osaka (IFO) 13693, 13772, 13773, 14815, and 15237] were screened for their BC production. The yield of the BC among all the strains from both the MO and MOL media was much higher than that from the HS medium. Acetobacter xylinum IFO 13772 was the best BC producer for all media. Furthermore, physical properties of these BC from the HS, MO, and MOL media were studied using Fourier-transform infrared spectroscopy, X-ray diffractometer, and cross polarization/magic angle spinning C-13 nuclear magnetic resonance. There are no significant differences in the crystallinity and the recorded I (alpha) fraction among the BC produced from the different media. A remarkable difference was only recorded in terms of viscosity. These results indicate that MO is a better carbon source than glucose for most of the strains investigated.