In the present work, actinomycete isolates with potential antibacterial activity were sought from animal manure. Twenty-two actinomycete isolates were isolated and nominated on the basis of their cultural properties on starch nitrate agar medium. The antibacterial activity of these isolates against a variety of bacterial and fungal species was examined. Actinomycete isolate A3 showed the highest antibacterial activity among those examined. The morphological, physiological, biochemical, and molecular characteristics of isolate A3 indicate that it belongs to the strain Streptomyces marokkonensis LMG 23016. The optimal conditions for development and antibacterial activity were submerged cultivation at 30 degrees Celsius and a pH of 7 for seven days. The optimal carbon and nitrogen sources are starch and potassium nitrate, respectively, and 1 % (w/v) sodium chloride. The antimicrobial metabolites of isolate A3 were extracted and analyzed using thin-layer chromatography (TLC), which revealed a solitary spot. UV, IR, HPLC, and GC/MS analysis confirmed diisooctyl phthalate (1,2-benzene dicarboxylic acid, diisooctyl ester) to be the major component. The results of the minimum inhibitory concentration and minimum bactericidal concentration tests on the S. marokkonensis strain LMG 23016 A3 extract demonstrated a broad-spectrum action against bacteria, thereby creating new opportunities for the development of potent antibiotic candidates.