4.5 Article

Purification, characterization and in vitro cytotoxicity of the bacteriocin from Pediococcus acidilactici K2a2-3 against human colon adenocarcinoma (HT29) and human cervical carcinoma (HeLa) cells

Journal

WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
Volume 27, Issue 4, Pages 975-980

Publisher

SPRINGER
DOI: 10.1007/s11274-010-0541-1

Keywords

Bacteriocin; Purification; Characterization; Cytotoxicity; Pediocin; Pediococcus acidilactici

Funding

  1. Philippine Council for Advanced Science and Technology Research and Development of the Department of Science and Technology, Philippines
  2. Toshiba Foundation International

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A bacteriocinogenic lactic acid bacterium (designated K2a2-3) isolated from the intestine of Philippine water buffalo was identified as Pediococcus acidilactici by 16S rRNA gene sequence analysis. The bacteriocin was purified by hydrophobic interaction chromatography, cation-exchange chromatography and reverse phase-high performance liquid chromatography. The purified protein has a molecular mass of 4,625.91 Da, quantified by electrospray ionization time-of-flight mass spectrometry. Based on a BLAST homology search of a partial sequence of 39 amino acid residues and the presence of the structural gene papA, detected through polymerase chain reaction, it was identified as very similar to pediocin PA-1. It was active against a wide spectrum of lactic acid bacteria and Listeria innocua. Partially-purified bacteriocin samples, conducted using pH-mediated bacteriocin extraction method, were found to be cytotoxic against human colon adenocarcinoma (HT29) and human cervical carcinoma (HeLa) cells in vitro, as determined by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay.

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