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Objectives:The current research is aimed to investigate the natural antimicrobial potential of Durio zibethinus ethanol leaves extracts (DZL).Methods: DZL was subjected to the preliminary phytochemical screening along with a quantitative analysis of phenols and flavonoids. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were recorded. The agar well diffusion method was used to measure the antibacterial activity against Gram-positive and Gram-negative bacteria. The microorganisms used in the study were ATCC strains of Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus, Streptococcus pyogenes, Neisseria gonorrhoeae, Pseudomonas aeruginosa, and Escherichia coli.Results: DZL exhibited the highest MIC of 0.1 mg/ml and MBC of 0.25 mg/ml against Gram-negative bacteria, P. aeruginosa and E. coli. At MIC of 0.1 mg/ml, DZL displayed significant zone of inhibition against P. aeruginosa and E. coli compared to gentamycin. Conclusion:This research has shown that DZL has natural antibacterial properties against Gram-negative human pathogens.