This study was carried out in Khartoum state, during November, 2019. Acacia nilotica was chosen for this study because of its using traditionally in treatment of many diseases. The Phytochemical constitutions of A. nilotica were investigated with aim to identify the antimicrobial effects of this plant. The dried fruit cover of Acacia nilotica was extracted successively with petroleum-ether, chloroform, methanol and distilled water. The phytochemical screening carried out on different extracts of Acacia nilotica fruit cover showed high amount of Glycosides, Flavonoids and Terpenoids (in all extracts) and moderate amount of Tannin in methanol and distilled water extracts, Alkaloids (acid and base) in methanol extract and Saponin in methanol and petroleum-ether extracts . The antimicrobial activity of extracts were evaluated against four standard bacteria species (gram positive bacteria; Staphylococcus aureus, Bacillus subtilis) and (gram negative bacteria; Pseudomonas aeruginosa, Escherichia coli). The plates were inoculated for sensitivity testing, minimum inhibitory concentration (MIC) was measured. The results of antimicrobial investigation show that the distilled water and methanolic extracts inhibited the growth of all microorganisms (Specified by the zone of inhibition). The results provide promising baseline information for potential use of these crude extracts in drug development programs in the pharmaceutical industries.