The aim of the present study was to investigate structural modifications and physicochemical properties of bacterial nanocellulose (BNC) pellicles produced statically in a beetroot peel medium as a mixture of C-sources (8% sucrose, 2% glucose, and 0.86% fructose). Characteristics of these BNC samples were compared to those produced using a chemically defined and modified Hestrin–Schramm medium with 3% (w/v) glucose, fructose, and sucrose as sole carbon sources. Compared with the beetroot peel medium, where 11.57 g/L BNC (wet weight) was obtained, glucose gave the highest BNC yield (13.07 g/L), sucrose 10.55 g/L, and fructose 7.9 g/L. FT-IR spectra showed almost identical chemical profiles assigned to cellulose I. XRD analysis revealed that the BNC produced in beetroot peel medium had a larger crystallite size than other BNC samples. The crystallinity index of beetroot peel BNC was higher than that of Hestrin–Schramm BNC. The water-holding capacity, water release rate, and iodine sorption value from the beetroot peel medium were greater than those of other BNCs. Furthermore, as there is a necessity today to find cheaper carbon sources to obtain valuable products at a lower cost, beetroot peels represent an interesting alternative to producing BNC as a new functional material for industrial applications.