Abstract: In the present study, the soil samples were collected from Corn and Soya fields; near MCAS. Actinomycetes strains were isolated in specific medium using Arginine-Glycerol-Salt (AGS). These Actinomycetes were screened with regard to potential against Gram-positive and Gram-negative bacteria. The purified Actinomycetes strains were performed in biochemical tests such as, Fermentation of citrate, Starch hydrolysis, Vogus-proskauer, Triple sugar iron test etc. The positive isolate was immobilized using sodium alginate beads and the immobilized cells were cultivated in specific fermentation liquid medium for 120 h. Antibacterial compound was purified from the filtrate by solvent extraction method. A comparative study on the total antibiotic sensitivity of the free cells and immobilized cells showed that the immobilized strains were found to be effective against the tested microorganisms. The immobilized cell of Actinomycetes was found to be more efficient for the production of secondary metabolites with batch fermentation. The Actinomycetes strains were prepared in crude protein and the crude protein was determined by using SDS PAGE method. From the results, we are concluded that the positive immobilized Actinomycetes strain showed high antibacterial activity against tested pathogens.