D-Glucose was converted into the 1,2:5,6-di-O-isopropylidene-α-D-glucofuranose (1) in good yield. Removal of 5,6-O-isopropylidene group from diacetone glucose (1) was achieved by careful hydrolysis which provided the monoacetal glucose (2). Here we reported the selective acylation of 1,2-O-isopropylidene-α- D-glucofuranose (2) by the direct method using a number of acylating agents furnished the corresponding 3,5,6-tri-O-acyl derivatives in reasonable yields. The structures of the newly synthesized compounds were ascertained by FTIR, 1H-NMR spectroscopy and elemental analysis. All the synthesized compounds were employed as test chemicals for in vitro antibacterial functionality test against six human pathogenic bacteria. The evaluation study revealed that the tested chemicals exhibited moderate to good antibacterial activities. It was also observed that the test chemicals were more effective against Gram-positive bacteria than that of the Gram-negative microorganisms.