Abstract: Background: Medicinal plants are now used as antibacterial compound that is safe for human use and overcome the multidrug resistant phenomenon. Objective: This study was done to compare the antibacterial activity of two fixed oils, purslane and MO. Methodology: The antimicrobial activity was done using oil diffusion method. The GC-MS analysis was done for the effective oil and protein profile and TEM scanning was done for the highly affected bacteria. Results: Purslane oil was the most effective especially against E. coli. Protein profile and TEM scanning revealed that the effect was on the outer bacterial membrane with no change in protein profile. The GC-MS analysis revealed that the oil contained Omega-3 fatty acid α-linolenic (ALA, C18:3 n-3); 9, 12, 15-octadecatrienoic acid methyl ester (30.83%). Conclusion: These fatty acids are bactericidal compounds that could disrupt the outer bacterial cell membrane and do not affect the internal protein content.