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

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First Report of an Anti- Peptide Produced by , Page 1 of 2

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Abstract:

The chapter describes the partial characterization of a new bacteriocin, produced by RK, active against . A bacterial strain, isolated from the environment, displayed an anti- 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 RK by a spot on lawn assay. The anti- 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- 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- bacteriocin. The anti- molecule is therefore an original peptide.

Citation: Héchard Y, Ferraz S, Bruneteau E, Steinert M, Berjeaud J. 2006. First Report of an Anti- Peptide Produced by , 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), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch96

Key Concept Ranking

Legionella pneumophila
0.6361617
Legionella longbeachae
0.605577
0.6361617
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Figures

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FIGURE 1

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

Citation: Héchard Y, Ferraz S, Bruneteau E, Steinert M, Berjeaud J. 2006. First Report of an Anti- Peptide Produced by , 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), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch96
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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. 64:31273133.
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: 24202425.
3. Fields, B. S.,, R. F. Benson, and, R. E. Besser. 2002. Legionella and Legionnaires’ disease: 25 years of investigation. Clin. Microbiol. Rev. 15:506526.
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: 44334444.
5. Molofsky, A. B., and, M. S. Swanson. 2004. Differentiate to thrive: lessons from the Legionella pneumophila life cycle. Mol. Microbiol. 53:2940.
6. Muder, R. R.,, V. L. Yu, and, A. H. Woo. 1986. Mode of transmission of Legionella pneumophila. A critical review. Arch. Intern. Med. 146:16071612.
7. Riley, M. A., and, J. E. Wertz. 2002. Bacteri-ocins: evolution, ecology, and application. Annu. Rev. Microbiol. 56:117137.
8. Steinert, M.,, U. Hentschel, and, J. Hacker. 2002. Legionella pneumophila: an aquatic microbe goes astray. FEMS Microbiol. Rev. 26:149162.

Tables

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TABLE 1

Antibacterial spectrum of raw extracts, assessed by well diffusion assays

Citation: Héchard Y, Ferraz S, Bruneteau E, Steinert M, Berjeaud J. 2006. First Report of an Anti- Peptide Produced by , 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), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch96

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