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Purification and Characterization of a New Bacillus thuringiensis Bacteriocin Active Against Listeria monocytogenes, Bacillus cereus and Agrobacterium tumefaciens

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Abstract

This study reports on the identification, characterization and purification of a new bacteriocin, named Bacthuricin F103, from a Bacillus thuringiensis strain BUPM103. Bacthuricin F103 production began in the early exponential phase and reached a maximum in the middle of the same phase. Two chromatographic methods based on high performance liquid chromatography and fast protein liquid chromatography systems were used to purify Bacthuricin F103. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis revealed that this bacteriocin had a molecular weight of approximately 11 kDa. It also showed a wide range of thermostability of up to 80 °C for 60 min and a broad spectrum of antimicrobial activity over a pH range of 3.0–10.0. This bacteriocin was noted, and for the first time, to exhibit potent antimicrobial activity against Agrobacterium subsp. strains, the major causal agents of crown gall disease in tomato and vineyard crops, and against several challenging organisms in food, such as Listeria monocytogenes and Bacillus cereus. Complete killing with immediate impact on cells was observed within a short period of time. The sequence obtained for Bacthuricin F103 by direct N-terminal sequencing shared considerable homology with hemolysin. Bacthuricin F103 was noted to act through the depletion of intracellular ions, which suggest that the cell membrane was a possible target to Bacthuricin F103.

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Acknowledgements

This work was supported by grants from the Tunisian Ministry of Higher Education, Scientific Research and Technology. The ‘Embassy of France’ at Tunis, Tunisia also provided a fellowship to Fakher Kamoun for a 3-month training in the U.G.M.E. INRA, La Minière, Versailles, France. We would like to express our gratitude to Mr. Mohamed Zribi for the supply of PALCAM medium. Special thanks are also due to Mr. Anouar Smaoui from the English Unit at the Sfax Faculty of Science for carefully proofreading and editing the manuscript of the present study.

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Authors and Affiliations

  1. Biopesticides Team (LPIP), Centre of Biotechnology of Sfax, P.O. Box ‘1177’, 3018, Sfax, Tunisia

    Fakher Kamoun, Ines Ben Fguira & Najeh Belguith Ben Hassen

  2. Faculty of Science, Soukra Road km 4, 3038, Sfax, Tunisia

    Hafedh Mejdoub

  3. Unité Génétique Microbienne et Environnement (UGME), INRA, La Minière, Versailles, France

    Didier Lereclus

  4. Biological and Environmental Sciences Department, College of Arts and Sciences, Qatar University, P.O. Box ‘2713’, Doha, Qatar

    Samir Jaoua

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  1. Fakher Kamoun
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  2. Ines Ben Fguira
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  3. Najeh Belguith Ben Hassen
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  4. Hafedh Mejdoub
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  5. Didier Lereclus
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  6. Samir Jaoua
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Correspondence to Samir Jaoua.

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Kamoun, F., Fguira, I.B., Hassen, N.B.B. et al. Purification and Characterization of a New Bacillus thuringiensis Bacteriocin Active Against Listeria monocytogenes, Bacillus cereus and Agrobacterium tumefaciens . Appl Biochem Biotechnol 165, 300–314 (2011). https://doi.org/10.1007/s12010-011-9252-9

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  • DOI: https://doi.org/10.1007/s12010-011-9252-9

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