Aim: To evaluate the antimicrobial potential of the methanolic extract of the fruit pulp of Hyphaene thebaica (Doum), a famous natural product in Sudan. Methods: Agar-well diffusion test and minimum inhibition diffusion test (MIC). Results: The investigation revealed that the extract showed no antifungal effect against Aspergillus niger ATCC 6275 and Candida albicans ATCC 10231. While it exhibited remarkable antibacterial activity against Staphylococcus aureus ATCC BAA 1026 and Pseudomonas aeruginosa ATCC 10145. Whereas, weak antibacterial activity was recorded with Bacillus cereus ATCC 10876 and Escherichia coli ATCC 1637. Conclusion: The fruit pulp of Hyphaene thebaica (Doum) has good antibacterial activity against some Gram-positive and Gram-negative bacteria. Accordingly, the consumption of this fruit would exert several beneficial effects by virtue of its antibacterial activity, and further studies using different bacterial strains and clinical pathogens are recommended.