The present work was conducted for characterization of antagonistic Bacillus sp. LBF–01 and its control mechanisms against 9 pathogenic fungi. The antifungal strains LBF–01 formed the highest inhibition zone diameters of 21mm – 12mm against the fungal strains in PDA media and exhibited 78% inhibition in NA media followed by 89% PDA and MEA, 67% CMA and 56% OMA media against A. alternata, C. gloeosporioides, F. moliniforme, F. oxysporum, Fusarium sp., A. parasiticus, and C. albicans indicating broad spectrum antifungal activity. Microscopic evaluation of the strain against treated Fusarium oxysporum showed that the antifungal secondary metabolites affect on growing mycelia in terms of swelling, cytoplasm’s coagulation and lysis of the mycelia. The strain is sensitive to common antibiotics used for human therapy indicating their non–virulent nature.