First Report of an Anti-Legionella Peptide Produced by Staphylococcus warneri

Yann Héchard; Sébastien Ferraz; Emilie Bruneteau; Michael Steinert; Jean-Marc Berjeaud

doi:10.1128/9781555815660.ch96

Chapter 96 : First Report of an Anti-Legionella Peptide Produced by Staphylococcus warneri

Authors: Yann Héchard¹, Sébastien Ferraz², Emilie Bruneteau³, Michael Steinert⁴, Jean-Marc Berjeaud⁵

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Affiliations: 1: Equipe de Microbiologie, Laboratoire de Chimie de l’Eau et de l’Environnement, UMR CNRS 6008, Université de Poitiers, 40 avenue du recteur Pineau, 86022 Poitiers, France; 2: Equipe de Microbiologie, Laboratoire de Chimie de l’Eau et de l’Environnement, UMR CNRS 6008, Université de Poitiers, 40 avenue du recteur Pineau, 86022 Poitiers, France; 3: Equipe de Microbiologie, Laboratoire de Chimie de l’Eau et de l’Environnement, UMR CNRS 6008, Université de Poitiers, 40 avenue du recteur Pineau, 86022 Poitiers, France; 4: Institut für Molekulare Infektionbiologie, Universität Wurzburg, 97070 Wúrzburg, Germany; 5: Equipe de Microbiologie, Laboratoire de Chimie de l’Eau et de l’Environnement, UMR CNRS 6008, Université de Poitiers, 40 avenue du recteur Pineau, 86022 Poitiers, France

Content Type: Monograph

Publication Year: 2006

Category: Bacterial Pathogenesis

Book DOI: 10.1128/9781555815660

Chapter DOI: 10.1128/9781555815660.ch96

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Abstract:

The chapter describes the partial characterization of a new bacteriocin, produced by Staphylococcus warneri RK, active against Legionella pneumophila. A bacterial strain, isolated from the environment, displayed an anti-Legionella activity by a spot on lawn assay. Various bacterial strains, gram-positive and gram-negative, were used as target organisms to test their sensitivity to S. warneri RK by a spot on lawn assay. The anti-Legionella molecule was then purified from the raw extract during a three-step purification procedure. First, the raw extract was diluted (50:50) with sodium acetate buffer (20 mM, pH 5) and applied to a weak cation exchange chromatography column (Hiprep 16/10 Carboxy-Methyl FF, Amersham Biosciences). Second, the latter fraction was applied onto a solid phase extraction C18 cartridge (Sep-pak plus, Waters), washed successively with 5 ml of 0, 10, 20, 30, 40, and 80% acetonitrile containing 0.1% trifluoroacetic acid. Third, the active fraction (80% acetonitrile) was lyophilized, solubilized with 1 ml of 40% acetonitrile, and injected on a Kromasil C8 reverse-phase HPLC analytical column (5μm, 100 Å, 4.6X250 mm, A.I.T.). The anti-Legionella activity has been eluted, during the last step, at 100% acetonitrile, indicating that the corresponding peptide is highly hydrophobic. This anti-Legionella-active fraction was subjected to mass spectrometry analysis, giving two molecular masses of 2613.8 Da and 2449.4 Da. This result shows that two peptides were present in the fraction, one of which likely corresponded to the anti-Legionella bacteriocin. The anti-Legionella molecule is therefore an original peptide.

Citation: Héchard Y, Ferraz S, Bruneteau E, Steinert M, Berjeaud J. 2006. First Report of an Anti-Legionella Peptide Produced by Staphylococcus warneri, p 411-413. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), Legionella. ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch96

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First Report of an Anti-Legionella Peptide Produced by Staphylococcus warneri

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/content/10.1128/9781555815660.ch96.ch96fig01

FIGURE 1

Spot on lawn assay. L. pneumophila Lens was spread on BCYE plate before spotting 5 μl of S. warneri RK culture. After 3 days of incubation at 37°C, a zone of inhibition of Legionella growth was noticed around the S. warneri RK colony.

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FIGURE 1

References

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1. Abu Kwaik, Y.,, L. Y. Gao,, B. J. Stone,, C. Venkataraman, and, O. S. Harb. 1998. Invasion of protozoa by Legionella pneumophila and its role in bacterial ecology and pathogenesis. Appl. Environ. Microbiol. 64: 3127– 3133.

2. Barker, J.,, M. R. Brown,, P. J. Collier,, I. Far-rell, and, P. Gilbert. 1992. Relationship between Legionella pneumophila and Acanthamoeba polyphaga: physiological status and susceptibility to chemical inactivation. Appl. Environ. Microbiol. 58: 2420– 2425.

3. Fields, B. S.,, R. F. Benson, and, R. E. Besser. 2002. Legionella and Legionnaires’ disease: 25 years of investigation. Clin. Microbiol. Rev. 15: 506– 526.

4. Kim, B. R.,, J. E. Anderson,, S. A. Mueller,, W. A. Gaines, and, A. M. Kendall. 2002. Literature review: efficacy of various disinfectants against Legionella in water systems. Water Res. 36: 4433– 4444.

5. Molofsky, A. B., and, M. S. Swanson. 2004. Differentiate to thrive: lessons from the Legionella pneumophila life cycle. Mol. Microbiol. 53: 29– 40.

6. Muder, R. R.,, V. L. Yu, and, A. H. Woo. 1986. Mode of transmission of Legionella pneumophila. A critical review. Arch. Intern. Med. 146: 1607– 1612.

7. Riley, M. A., and, J. E. Wertz. 2002. Bacteri-ocins: evolution, ecology, and application. Annu. Rev. Microbiol. 56: 117– 137.

8. Steinert, M.,, U. Hentschel, and, J. Hacker. 2002. Legionella pneumophila: an aquatic microbe goes astray. FEMS Microbiol. Rev. 26: 149– 162.

Tables

First Report of an Anti-Legionella Peptide Produced by Staphylococcus warneri

/content/10.1128/9781555815660.ch96.ch96tab01

TABLE 1

Antibacterial spectrum of S. warneri raw extracts, assessed by well diffusion assays

TABLE 1

Antibacterial spectrum of S. warneri raw extracts, assessed by well diffusion assays

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