Screening, characterisation and production of bacteriocin from Klebsiella pneumoniae

<p>Bacteriocins are antimicrobial peptides produced by bacteria to control growth of rival strains of bacteria. A potential use of bacteriocins is as an alternative antibiotic, as they usually have narrow specificity, pathogenic bacteria can be eradicated without disturbing the normal microbiome. The aim of this project was to distinguish bacteriocin producing strains of Klebsiella pneumoniae from nosocomial samples, characterise the bacteriocins and use molecular cloning to allow large scale production of bacteriocin without use of pathogenic bacteria.Bacteriocin producing strains of bacteria were identified by their inhibition of sensitive strains in an agar overlay. A selection of bacteriocins produced were chosen to provide a range of activity, and were then first purified by ion exchange chromatography, then size exclusion chromatography, followed by SDS-PAGE. Once isolated, bacteriocins from three strains were analysed by MALDI MS/MS.Primers were designed for a previously identified klebicin, allowing amplification of the bacteriocin genes by PCR and cloning into non-pathogenic BL21 (DE3) E. coli. After being cloned into BL21 (DE3) E. coli, the production of purified bacteriocin without use of pathogenic bacteria was possible.This allows future investigations into the possible applications of bacteriocins, such as alternative antibiotics, water or air filtration, or as components in bacterial contamination detecting devices.</p>