The aim of this study was isolation and characterization of antimicrobial metabolite producing actinomycetes from forest soil. A new actinomycete designated ERIAS-10 was selected based on antibacterial and antifungal activity in the primary screening. Biochemical, physiological, morphological and carbohydrate fermentation pattern of the isolate was characterized and confirmed that the isolate belong to Strepomyces species. The 16S rRNA gene sequence shared 100% sequence similarity to Streptomyces roseoverticillatus. The antimicrobial substances were extracted with organic solvents from the spent medium in which ERIAS-10 was cultivated. Antimicrobial activity was assessed using the broth micro dilution technique. The minimum inhibitory concentration (MIC) of the extracts against Enterococcus faecalis was 7.81 µg/mL and Aspergillus niger, A. flavus, A. clavatus and Fusarium oxysporum was 15.63 and 31.25 µg /mL respectively, whereas, Humicola grisea showed MIC at 150 µg/mL level. Cloning and sequence analysis of ketoacyl synthase gene revealed similarity to the type II polyketide synthase gene of Streptomyces species. The results showed that the antimicrobial potential of ERIAS-10 isolate could be suitable for pharmaceutical applications.