Production of bacterial cellulose (BC) by Gluconacetobacter xylinus was investigated in static culture form, using glycerol and biodiesel residue as the carbon sources and as raw material for technological purposes. The cellulosic material, obtained as a membrane, was characterized by spectroscopic techniques and its morphological aspects were investigated by scanning electron microscopy (SEM). In a medium containing 3% (w/v) glycerol, the BC yield was higher than that from a glucose and mannitol based-medium. After 10 days, using a medium containing biodiesel residue, we obtained a satisfactory amount of BC. FT-IR spectra revealed that all the BC samples showed characteristic peaks, associated with the usual functional groups for this cellulosic material. These results suggested that glycerol could be a potential low-cost substrate for BC industrial scale production, based on its high performance and economic viability.