A total of two marine actinomycetes were isolated from different locations of the Manakudi Estuary of Arabian Sea in Tamilnadu, India. They exhibited higher antagonistic activity against the Gram positive bacteria; methicillin resistant and susceptible Staphylococcus aureus, Enterobacter sp, Salmonella typhi, Bacillus subtilis, Klebsiella pneumoniae, Proteus vulgaris. For Gram negative organism Pseudomas auregionosa it showed intermediate activity and no antagonistic effect towards yeast like Candida albicans. Pink actinomycetes (PJS) with white aerial mycelium and pink substrate mycelium and black colonies (BJS) of white aerial mycelium and yellowish white substrate mycelium showed potent inhibiting effect of other microorganisms. Biochemical analysis of PJS and BJS revealed these organisms are Gram positive, starch, mannose, glucose, sucrose, fructose, lipase and urease positive, amylase, lipase enzyme positive and urease negative. All the isolated actinomycetes were resistant to nalidixic acid, methicillin and penicillin. 16S rDNA phylogenetic typing gave ~1500 bp amplified product and it was cloned in pGEMT easy vector. Sequencing of amplified product will give the phylogeny of isolated actinomycetes and the further study on this organism may provide a new antibiotic for the welfare of human being.