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Effect of a highly concentrated lipopeptide extract of Bacillus subtilis on fungal and bacterial cells

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Abstract

Lipopeptides produced by Bacillus subtilis are known for their high antifungal activity. The aim of this paper is to show that at high concentration they can damage the surface ultra-structure of bacterial cells. A lipopeptide extract containing iturin and surfactin (5 mg mL−1) was prepared after isolation from B. subtilis (strain OG) by solid phase extraction. Analysis by atomic force microscope (AFM) showed that upon evaporation, lipopeptides form large aggregates (0.1–0.2 μm2) on the substrates silicon and mica. When the same solution is incubated with fungi and bacteria and the system is allowed to evaporate, dramatic changes are observed on the cells. AFM micrographs show disintegration of the hyphae of Phomopsis phaseoli and the cell walls of Xanthomonas campestris and X. axonopodis. Collapses to fungal and bacterial cells may be a result of formation of pores triggered by micelles and lamellar structures, which are formed above the critical micelar concentration of lipopeptides. As observed for P. phaseoli, the process involves binding, solubilization, and formation of novel structures in which cell wall components are solubilized within lipopeptide vesicles. This is the first report presenting evidences that vesicles of uncharged and negatively charged lipopeptides can alter the morphology of gram-negative bacteria.

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Abbreviations

AFM:

Atomic force microscope

NRPS:

Non-ribosomal peptide synthetase

OM:

Optical microscopy

PKS:

Polyketide synthase

PE:

Plant emergence

PH:

Plant height

PDW:

Plant dry weight

Xac:

Xanthomonas axonopodis pv. citri

Xcc:

Xanthomonas campestris pv. campestris

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Acknowledgments

This research has been funded by the Fundação de Pesquisa de São Paulo (Fapesp grants 96/11193-7 and 03/12529-4) and the Conselho Nacional de Pesquisa (CNPq). We would like to thank the valuable research assistance of L. O. Bonugli and J. R. Castro. M. E. Silva-Stenico was the recipient of postdoctoral fellowship from FAPESP (Grant 2004/16042-5)

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Authors and Affiliations

  1. Faculdade de Química, PUC-Campinas, C. Postal 317, Campinas, SP, 13012-970, Brazil

    Augusto Etchegaray & Carolina de Castro Bueno

  2. Laboratório de Microbiologia, Embrapa Meio Ambiente, Jaguariuna, SP, Brazil

    Itamar Soares de Melo

  3. CENA, Universidade de São Paulo, C. Postal 96, Piracicaba, SP, 13400-970, Brazil

    Siu Mui Tsai, Marli de Fátima Fiore & Maria Estela Silva-Stenico

  4. Departamento de Química, Universidade de São Paulo, Av. Bandeirantes, Ribeirão Preto, SP, 14100-000, Brazil

    Luiz Alberto Beraldo de Moraes

  5. Laboratório de Nanoestruturas e Interfaces, Instituto de Física, UNICAMP, Campinas, SP, 13083-970, Brazil

    Omar Teschke

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  1. Augusto Etchegaray
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  2. Carolina de Castro Bueno
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  3. Itamar Soares de Melo
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  4. Siu Mui Tsai
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  6. Maria Estela Silva-Stenico
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  7. Luiz Alberto Beraldo de Moraes
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  8. Omar Teschke
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Correspondence to Augusto Etchegaray.

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Communicated by Jorge Membrillo-Hernández.

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Etchegaray, A., de Castro Bueno, C., de Melo, I.S. et al. Effect of a highly concentrated lipopeptide extract of Bacillus subtilis on fungal and bacterial cells. Arch Microbiol 190, 611–622 (2008). https://doi.org/10.1007/s00203-008-0409-z

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  • DOI: https://doi.org/10.1007/s00203-008-0409-z

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