Abstract
Multi- and pan-antibiotic-resistant bacteria are a 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.
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Acknowledgments
The authors would like to thank Nicholas Stella for technical assistance and Dr. Yohei Doi, Division of Infectious Diseases, University of Pittsburgh School of Medicine for kindly providing bacterial strains. This work was supported by funding from the Foundation of UMDNJ faculty research grant to D.E.K., and NIH AI085570 and a Research to Prevent Blindness Career Development Award to R.M.Q.S. Additional support was provided by NIH grant EY08098 and the Eye and Ear Foundation of Pittsburgh.
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Communicated by Erko Stackebrandt.
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Shanks, R.M.Q., Dashiff, A., Alster, J.S. et al. Isolation and identification of a bacteriocin with antibacterial and antibiofilm activity from Citrobacter freundii . Arch Microbiol 194, 575–587 (2012). https://doi.org/10.1007/s00203-012-0793-2
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DOI: https://doi.org/10.1007/s00203-012-0793-2