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Purification and Characterization of a Novel Anti-Campylobacter Bacteriocin Produced by Lactobacillus curvatus DN317

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Abstract

The lactic acid bacteria (LAB) microbiota of Saudi chicken ceca was determined. From 60 samples, 204 isolates of lactic acid bacteria were obtained. Three isolates produced antimicrobial activities against Campylobacter jejuni, Listeria monocytogenes, and Bacillus subtilis. The isolate DN317, which had the highest activity against Campylobacter jejuni ATCC 33560, was identified as Lactobacillus curvatus (GenBank accession numbers: KX353849 and KX353850). Full inhibitory activity was observed after a 2-h incubation with the supernatant at pH values between 4 and 8. Only 16% of the activity was conserved after a treatment at 121 °C for 15 min. The use of proteinase K, pepsin, chymotrypsin, trypsin, papain, and lysozyme drastically reduced the antimicrobial activity. However, lipase, catalase, and lysozyme had no effect on this activity. The active peptide produced by Lactobacillus curvatus DN317 was purified by precipitation with an 80% saturated ammonium sulfate solution, and two steps of reversed phase HPLC on a C18 column. The molecular weight of this peptide was 4448 Da as determined by MALDI-ToF. N-terminal sequence analysis using Edman degradation revealed 47 amino acid residues (UniProt Knowledgebase accession number C0HK82) revealing homology with the amino acid sequences of sakacin P and curvaticin L442. The antimicrobial activity of the bacteriocin, namely curvaticin DN317, was found to be bacteriostatic against Campylobacter jejuni ATCC 33560. The use of microbial antagonism by LAB is one of the best ways to control microorganisms safely in foods. This result constitutes a reasonable advance in the antimicrobial field because of its potential applications in food technology.

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Acknowledgments

This work was supported by the deanship of Research, King Faisal University.

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Authors and Affiliations

  1. Unité de Protéomique Fonctionnelle et Potentiel Nutraceutique de la Biodiversité de Tunisie, Institut Supérieur des Sciences Biologiques Appliquées de Tunis, Rue Z. Essafi, 1006, Tunis, Tunisia

    Mohamed Zommiti & Mounir Ferchichi

  2. Department of Medical Microbiology and Parasitology, College of Medicine, King Faisal University, P.O. Box: 400, Al-Ahsa, 31982, Saudi Arabia

    Hamdan Almohammed

  3. College of Applied Medical Sciences, Clinical Laboratory Department, King Faisal University, P.O. Box: 401, Al-Ahsa, 31982, Saudi Arabia

    Mounir Ferchichi

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  1. Mohamed Zommiti
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Correspondence to Mounir Ferchichi.

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All Institutional and National Guidelines for the care and use of chickens were followed.

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Zommiti, M., Almohammed, H. & Ferchichi, M. Purification and Characterization of a Novel Anti-Campylobacter Bacteriocin Produced by Lactobacillus curvatus DN317. Probiotics & Antimicro. Prot. 8, 191–201 (2016). https://doi.org/10.1007/s12602-016-9237-7

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