K Haritha, P Udayasri, J Madhavi, KK Pulicherla and KRS Sambasiva Rao
Keywords: Cationc synthetic peptide, immobilized metal affinity chromatography system, placket burman model, antibacterial activity.
Abstract: Cationic antimicrobial peptides are the upcoming therapeutic molecules as alternative drugs to the antibiotics. These peptides have a good scope in current antibiotic research. In the present study E. coli strain GJ1158 host was chosen for the expression of gene for Insilco designed synthetic peptide, Using Modified M9 medium. Various trails were carried out to optimize the recombinant peptide production in modified M9 medium by following the Placket Burman model. The optimal media was chosen for further expression studies and the expressed antimicrobial peptide was purified using Immobilized Metal Affinity Chromatography (IMAC) system. The product was visualized on 16% Tricine SDS-PAGE. It was identified that 30% of the bacterial proteins as the recombinant protein. The expressed antimicrobial peptide was purified using Immobilized Metal Affinity Chromatography (IMAC) system. The antimicrobial activity of purified peptide using Top agar assay showed that the recombinant antimicrobial peptide has high antibacterial activity against both Gram-positive and -negative bacteria.
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