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Purification and identification of the pediocin produced by Pediococcus acidilactici MM33, a new human intestinal strain
Author(s) -
Millette M.,
Dupont C.,
Shareck F.,
Ruiz M.T.,
Archambault D.,
Lacroix M.
Publication year - 2008
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2007.03583.x
Subject(s) - pediococcus acidilactici , bacteriocin , microbiology and biotechnology , pediococcus , biology , plasmid , listeria monocytogenes , antimicrobial , bacteria , biochemistry , lactobacillus , gene , lactic acid , fermentation , lactobacillus plantarum , genetics
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.