title: Isolation and identification of a bacteriocin with antibacterial and antibiofilm activity from Citrobacter freundii.
abstract: Multi- and pan-antibiotic-resistant bacteria area major health challenge in hospital settings. Furthermore,when susceptible bacteria establish surface-attached biofilm populations, they become recalcitrant to antimicrobial therapy. Therefore, there is a need for novel antimicrobials that are effective against multi-drug-resistant and surface-attached bacteria. A screen to identify prokaryote-derived antimicrobials from a panel of over 100 bacterial strains was performed. One compound isolated from Citrobacter freundii exhibited antimicrobial activity against a wide range of Gram-negative bacteria and was effective against biofilms. Random transposon mutagenesis was performed to find mutants unable to produce the antimicrobial compound.Transposons mapped to a bacteriocin gene located on a small plasmid capable of replication in Escherichia coli. The plasmid was sequenced and found to be highly similar to a previously described colicinogenic plasmid.Expression of the predicted bacteriocin immunity gene conferred bacteriocin immunity to E. coli. The predicted bacteriocin gene, colA-43864, expressed in E. coli was sufficient to generate anti-microbial activity, and purified recombinant ColA-43864 was highly effective in killing E. coli, Citrobacter species, and Klebsiella pneumoniae cells in a planktonic and biofilm state. This study suggests that bacteriocins can be an effective way to control surface-attached pathogenic bacteria.
authors.affiliation: Campbell Laboratory of Ophthalmic Microbiology, Department of Ophthalmology, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Robert M Q
authors.initials: RM
authors.lastname: Shanks
authors.forename: Aliza
authors.initials: A
authors.lastname: Dashiff
authors.forename: Jason S
authors.initials: JS
authors.lastname: Alster
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Kadouri
mesh_terms: D000900:Anti-Bacterial Agents / Q000302:isolation & purification* / Q000494:pharmacology*; D001430:Bacteriocins / Q000302:isolation & purification* / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects*; D016971:Citrobacter freundii / Q000737:chemistry*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D006090:Gram-Negative Bacteria / Q000187:drug effects* / Q000302:isolation & purification; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D001430:Bacteriocins; D011994:Recombinant Proteins