Regioselective acylation of monosaccharides are highly significant for the preparation of both natural and non-natural carbohydrate compounds and their synthetic intermediates. In this respect, an efficient simple method for site selective 6-O-hexanoylation of octyl β-D-glucopyranoside (OG) at low temperature is described. The 6-O-hexanoylglucoside, thus formed, was then treated with five acyl halides and obtained novel corresponding O-acylglupyranosides at C-2, C-3 and C-4 positions with high yields. Activity spectra prediction for substances (PASS) indicated that these glucoside esters are potential against antimicrobials especially against fungi. In vitro antibacterial test indicated them as weak to moderate inhibitor of Gram-positive organisms. ADMET prediction indicated that these OGs are overall safe to use. Density functional theory (DFT) optimized electronic energy, enthalpy (ΔH), Gibbs free energy, entropy, dipole-moment (μ), and molecular electrostatic potential (MEP) are also discussed.