In the present study a 9 amino acid synthetic anti microbial peptide was designed byusing the bioinformatics tools viz., ProtParam, Hierarchical Neural Network, Anti microbialpeptide database. The designed peptide was back translated to core nucleotide sequence and thegene for the peptide was constructed by over lapping PCR methodology. Synthesized genewas cloned into pRSET-A vector and further the peptide was expressed in E.coli host systems-BL (21) DE3 and GJ1158. A comparative study was performed and identified GJ1158 as the bestexpression system and the anti bacterial activity was analyzed on various Gram +Ve and Gram –Ve bacteria.