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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

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Abstract

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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Acknowledgments

This work was supported by an MS thesis grant from the Philippine Council for Advanced Science and Technology Research and Development of the Department of Science and Technology, Philippines. Part of the study was also made possible through the Asia in Today’s World Program (Kyushu University Summer Program) 2007 with the support of Toshiba Foundation International.

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Authors and Affiliations

  1. Department of Biology, School of Science and Engineering, Loyola Schools, Ateneo de Manila University, Katipunan Avenue, Loyola Heights, 1108, Quezon City, Philippines

    Karen I. Villarante & Evelyn E. de Guzman

  2. National Institute of Molecular Biology and Biotechnology, BIOTECH, University of the Philippines Los Baños, College, 4031, Laguna, Philippines

    Francisco B. Elegado

  3. Laboratory of Microbial Science and Technology, Division of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1, Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan

    Shun Iwatani, Takeshi Zendo & Kenji Sonomoto

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  1. Karen I. Villarante
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  2. Francisco B. Elegado
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  3. Shun Iwatani
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  4. Takeshi Zendo
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  5. Kenji Sonomoto
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  6. Evelyn E. de Guzman
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Correspondence to Francisco B. Elegado.

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Villarante, K.I., Elegado, F.B., Iwatani, S. et al. 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. World J Microbiol Biotechnol 27, 975–980 (2011). https://doi.org/10.1007/s11274-010-0541-1

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  • DOI: https://doi.org/10.1007/s11274-010-0541-1

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