In this study, the effect of modified Hestrin Schramm (HS) medium supplemented with different carbon sources viz., glucose, fructose, galactose and lactic acid on the yield and physicochemical properties of bacterial cellulose (BC) produced from Gluconacetobacter xylinus strain MTCC 7795 in shake flask culture conditions was investigated. Growth studies indicated that all carbon sources supported the growth of bacteria, though specific growth rate and doubling time differs. Fructose gave the highest cellulose yield of 7.72 mg/ml after 130 h of fermentation, while yield in glucose and galactose supplemented medium were 4.49 mg/ml and 3.38 mg/ml, respectively. X-ray powder diffraction (XRD) analysis revealed that all BC samples were amorphous in comparison to commercial cellulose. Fourier transform infrared (FTIR) spectroscopic investigations of bacterial cellulose (BC) samples affirm the purity of the cellulose produced. No significant variations in physicochemical properties of cellulose samples produced with different carbon sources were observed. This study for the first time has investigated the effect of carbon sources on physicochemical properties of bacterial cellulose produced by G. xylinus MTCC 7795 and provides a strategy for economical production of BC with anticipated application in therapeutics and tissue engineering.