The objectives of this study were analysis of the secondary metabolite products from Aspergillus fumigatus and evaluation antibacterial activity. Fifteen bioactive compounds were identified in the methanolic extract of Aspergillus fumigatus. The identification of bioactive chemical compounds is based on the peak area, retention time molecular weight and molecular formula. GC-MS analysis ofAspergillus fumigatus revealed the existence of the 9-Hexadecenoic acid ,2,4-Dimethyl-5-methylthiopent-4-en-2-ol , E-11-Hexadecenoic acid , ethyl ester , D-Glucose , 6-O-α-D-galactopyranosyl- , α-D-Glucopyranoside , O-α-D-glucopyranosyl-(1.fwdarw.3)-β- , 6-Acetyl –β-d-mannose , 4H-Pyran-4-one,2,3-dihydro-3,5- dihydroxy-6-methyl- , 5-Hydroxymethylfurfural , β-D-Glucopyranoside, methyl , Tetraacetyl-d-xylonic nitrile , n-Hexadecanoic acid , 9-Octadecenoic acid , (2-phenyl-1,3-dioxolan-4-yl)methyl ester , Octadecanoic acid ,9,10-Secocholesta-5,7,10(19)-triene-3,24,25-triol,(3β,5Z,7E)- and Pyrimidin-2-ol,4-(3,4- dimethoxyphenyl)-6-phenyl. Gramineae poaceae was very highly antifungal activity (8.32±0.09) mm while
Streptococcus faecalis has a maximum zone formation (6.92±0.24) mm against Aspergillus fumigatus.