Abstract: The research work was conducted to investigate the in vitro antimicrobial activity of the compound Aglycon of Deoxy-Niazimicine (N-benzyl,S-ethyl thioformate) isolated from Moringa oleifera Lam. in comparison with crude chloroform extract. Antibacterial and antifungal activities of the compound-1 and crude extract were observed against fourteen pathogenic bacteria and six pathogenic fungi. Compound-1 isolated from the chloroform (CHCl3) fraction of ethanol extract showed comparatively more in vitro antibacterial activity against Shigella boydii, Shigella dysenteriae and Staphylococcus aureus than the crude chloroform (CHCl3) extract and produced zone of inhibition in between 09 and 13 mm. The MIC values of the compound-1 and crude extract were also determined against B. megaterium, St. aureus, Sh. dysenteriae and Sh. boydii. The values were found to be between 32 to 128 μg ml-1. The compound-1 also showed moderate antifungal activity against Candida albicans and Aspergillus flavus than the crude chloroform extract although the crude extract showed moderate activity against Trichoderma species. Thus compound-1 has antibacterial and antifungal activity and suggested its therapeutic use as an antimicrobial agent.