Antibacterial agents are essential drugs for combating the bacterial infections. In this study, the potent actinomycetes strain active against pathogenic bacteria and fungi was isolated from Solapur region of Maharashtra. Based on morphological, physiological, biochemical and 16S rRNA sequence, isolate was identified as Streptomyces sp. JF714876. The nutritional requirement and production condition were optimized to increase the antibiotic production. The crude compound from optimized media was extracted using ethyl acetate at pH -3, the active fraction was purified and separated using silica gel column chromatography and thin layer chromatography with Rf value 0.8. The MIC of active compound tested against Aspergillus species was recorded at 40 μg/ml whereas against dermatophytes at 50 μg/ml concentration. Physicochemical characteristic of bioactive agent indicate that the compound belongs to alkaloid group. © 2023 International Journal of Pharmaceutical Sciences and Nanotechnology. All rights reserved.