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  • Title: Purification and identification of the pediocin produced by Pediococcus acidilactici MM33, a new human intestinal strain.
    Author: Millette M, Dupont C, Shareck F, Ruiz MT, Archambault D, Lacroix M.
    Journal: J Appl Microbiol; 2008 Jan; 104(1):269-75. PubMed ID: 17927747.
    Abstract:
    AIMS: The aim of this study was to purify and identify the bacteriocin produced by Pediococcus acidilactici MM33, a strain previously isolated from human gut. METHODS AND RESULTS: Purification of the bacteriocin was performed by cationic exchange chromatography followed by a reverse phase step. Biochemical and mass spectrometry analysis showed homology with pediocin PA-1. To verify if P. acidilactici MM33 carried the pediocin PA-1 gene, total DNA was used to amplify the pediocin gene. The PCR product obtained was then sequenced and the nucleotide sequence revealed to be identical to that of pediocin PA-1. Treatment of P. acidilactici MM33 with novobiocin resulted in a plasmid-cured strain without bacteriocin-producing capacity. Antimicrobial assay and molecular analysis demonstrated that this strain was ped(-) suggesting that the ped cluster is plasmid encoded. Antimicrobial assay revealed that pediocin was bactericidal against Listeria monocytogenes, showing a minimal inhibitory concentration (MIC) of 200 AU ml(-1). CONCLUSIONS: A two-step purification procedure was elaborated in this study. The bacteriocin secreted by the human strain P. acidilactici MM33 is carried on a plasmid and the amino acid sequence is identical to pediocin PA-1. SIGNIFICANCE AND IMPACT OF THE STUDY: Pediococcus acidilactici MM33 is the first human pediocin-producing strain reported and could be used as probiotic to prevent enteric pathogen colonization.
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