Antibiotics-resistant infections have appeared in numerous regions in the world, which draws attention to the urgent need for novel antimicrobial compounds. The current investigation aims to evaluate the activity of crude extracts from some actinomycetes isolates as potential antibacterial and antifungal agents. Fifteen actinomycetes were isolated from cultivated soil samples collected from various regions at Suez governorate. The antimicrobial properties of the crude extracts were tested against a variety of pathogenic bacteria and fungi. The selected ethyl acetate extract (EtOAc) of the isolate SUN1 showed strong antifungal activity against both Aspergillus fumigatus and Fusarium solani with inhibition zones 28 and 30 mm respectively, while showed moderate antibacterial activity against Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus “MRSA" with inhibition zones 14 and 18 mm respectively. Based on the physiological, biochemical, and microscopic characterization of the isolate SUN1, it was characterized and identified on the genus level as Streptomyces sp. The EtOAc extract of Streptomyces SUN1 was characterized using the principal GC-MS chemical fingerprint. The net GC-MS data revealed that the primary concentrations are represented by phenolic and oxygenated chemicals. Eleven bioactive compounds were identified and classified as the principal components. These compounds included tributyl acetyl citrate, 1,4-benzenedicarboxylic acid, bis(2-ethylhexyl) ester, bis(2-ethylhexyl) phthalate and butyl citrate with relative amounts of 34.81, 28.39, 10.25, and 10.08% respectively. The investigation indicates that the isolated actinomycetes offer a valuable source of antimicrobial compounds that can be applied as novel antibiotics.