Background: A fungus isolated from soil displayed considerable antibacterial activity. The fungus was identified as Stemphylium radicinum (Meier, Drechsler and Eddy, 1922) based on morphological characterization. Methods: Fungal extraction was carried out by ethyl acetate solvent. The metabolite showed Bioactivity against five bacterial strains. Esherichia coli from urine, Staphylococcus aureus from otitis media, Streptococcus pyogenes from Throat, Pseudomonas aeruginosa and Klebsiella pneumoniae from burns by using a disc diffusion technique was examined. Results: The inhibition zones exhibited by fungal extract were 14.0 - 26.0 mm MIC test revealed that the extract of S. radicinum exhibited a minimal inhibition values ranging between (3.12 - 12.5 ug/ml) and MBC ranging between (6.25 - 50.0 ug/ml) against bacterial strains. A verification of non- toxicity of the fungal extract against human blood revealed a negative test. The chemical analysis of the fungal crude extract showed that extract S. radicinum contains Tannins group, phenol compounds and amino acid, absent flavenoide. Conclusion: The metabolite produced by the fungi could be an alternative source of antimicrobial agents against clinical pathogens.