Antibiotics are heterogeneous group of small organic molecules obtained from microbial origin. At low concentrations they are effective against most of the microbial pathogens that cause infections in animals and humans. Over past two decades numerous antibiotics, like acidophilin, bulgaricin, colicins, natamycin, reuterin etc. have been produced by bacteria. Certain members of bacteria can develop drug resistance, which has been a major drawback in the field. This has paved way for the present study on isolation of antimicrobial compound producing bacteria. The bacterial cultures were obtained from soil of pine forest, Western ghats (Kodaikanal) and lake water. The isolate VITRAS3 was effective against all the tested pathogens. The isolates were mass multiplied for the production of antimicrobial compounds. Further the compound produced by the isolate was extracted using polar and non-polar solvents. The compound was identified using gas chromatography-mass spectrometry (GCMS). The bacteria obtained were capable of producing antimicrobial compound and effectively inhibited bacterial pathogens.