Nicotiana tabacum has been widely used as a therapeutic plant in Asian countries including China, India, Cambodia, Nepal, Malaysia and Iran. The leaves of Nicotiana tabacum have been reported to activate biological mechanisms such as antibacterial, antifungal, antimicrobial, anthelmintic and anti-Alzheimer’s activities. This study was conducted to investigate the proximate, antimicrobial and phytochemical composition of tobacco leaves (Nicotiana tabacum) extract. Nicotiana tabacum leaves were subjected to proximate analysis and the alcoholic extract was investigated for antimicrobial inhibition against a wide range of bacteria using agar well diffusion method. The extract was also investigated for phytochemical composition. The alcohol extract subjected to analyses was prepared using 95% ethanol as extracting solvent. The proximate analysis showed that the leaves of Nicotiana tabacum contain significantly high protein, carbohydrate and fibre while the fat, ash and moisture content were significantly low. Phytochemical analysis of the alcohol extract of