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
2011
Villarante, Karen I. | Elegado, Francisco B. | Iwatani, Shun | Zendo, Takeshi | Sonomoto, Kenji | de Guzman, Evelyn E.
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 4,625.91Â 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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