The main objective of the present study was isolation, purification, and characterization of actinomycetes from soil samples. Soil samples were taken from different niche habitats of Vellore district, Tamil Nadu, India. Potential isolate was screened morphologically and biochemically characterised, further were subjected to extraction for the creation of the antibacterial compound. The strains JS26, JS27, and JS28 were discovered profoundly potential, Isolate JS27 displayed the most noteworthy activity against Bacillus cereus (22 mm) and Enterococcus faecalis (25 mm). Ethyl acetate extract showed 89% restraint towards free radicals DPPH (1,1-Diphenyl-2-picrylhydrazyl). The gas chromatography-mass spectroscopy (GC-MS) demonstrated the few unstable mixes. Consequently the outcome showed that the ethyl acetate concentrate of confine groups have tendency to prevent cancer and could be utilized as a part of the advancement of new bioactive metabolites for pharmaceutical purposes. © RJPT All right reserved.