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Production of Anti-Methicillin-Resistant Staphylococcus Activity from Bacillus subtilis sp. Strain B38 Newly Isolated from Soil

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Abstract

B38 bacterial strain, isolated from Tunisian soil showed a strong antimicrobial activity. Based on biochemical characterization and 16S rDNA sequence analysis, B38 strain was identified as Bacillus subtilis. Cell culture supernatant showed antibacterial activity against clinical isolates of methicillin-resistant Staphylococcus species and several Gram-positive and Gram-negative bacteria. Antifungal activity against phytopathogenic fungi was also observed. Antibacterial activity production started at early exponential growth phase, and maximum activity was reached at the stationary phase. This antibacterial activity was neither affected by proteases, lipase, and organic solvents, nor by surfactants. It was stable over a wide pH range and still active after autoclaving at 121 °C during 20 min. Thin layer chromatography followed by bioautography assay allowed the detection of four active spots with R f values of 0.30, 0.47, 0.70, and 0.82. The single spot with R f 0.30 showed antifungal activity, whereas the spots with R f values of 0.47, 0.70, and 0.82 exhibited antibacterial activity.

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Abbreviations

AU:

Activity unit

CFU:

Colony forming unit

CS:

Culture supernatant

LB:

Luria–Bertani broth

MTT:

Methylthiazolyltetrazolium bromide

NB:

Nutrient broth

PCR:

Polymerase chain reaction

PDB:

Potato dextrose broth

rDNA:

ribosomal DNA

R f :

Retention factor

TCA:

Trichloroacetic acid

TLC:

Thin layer chromatography

TSB:

Trypticasein soy broth

TY:

Tryptic Yeast

UV:

Ultra violet

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Acknowledgements

This work was supported by grants from the “Ministère de l’Enseignement Supérieur, de la Recherche Scientifique et de la Technologie” of Tunisia. We thank Prof. E. Aouani for valuable discussion and critical reading of the manuscript.

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Authors and Affiliations

  1. Laboratoire Interactions Légumineuses-Microorganismes, Centre de Biotechnologie, Technopole Borj-Cedria, BP-901, 2050, Hammam-lif cedex, Tunisia

    Olfa Tabbene, Imen Ben Slimene, Faten Bouabdallah & Ferid Limam

  2. Unità di Diagnostica Molecolare Avanzata, II Facoltà di Medicina e Chirurgia, Azienda Ospedaliera S. Andrea, Roma, Italy

    Maria-Luisa Mangoni

  3. Laboratoire de Microbiologie, UMR 5248, ENITA de Bordeaux, BP-201, 33175, Gradignan, France

    Maria-Camino Urdaci

Authors
  1. Olfa Tabbene
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  2. Imen Ben Slimene
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  3. Faten Bouabdallah
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  4. Maria-Luisa Mangoni
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  5. Maria-Camino Urdaci
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  6. Ferid Limam
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Correspondence to Ferid Limam.

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Tabbene, O., Ben Slimene, I., Bouabdallah, F. et al. Production of Anti-Methicillin-Resistant Staphylococcus Activity from Bacillus subtilis sp. Strain B38 Newly Isolated from Soil. Appl Biochem Biotechnol 157, 407–419 (2009). https://doi.org/10.1007/s12010-008-8277-1

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  • DOI: https://doi.org/10.1007/s12010-008-8277-1

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