Novosphingobium sp. PP1Y as a novel source of outer membrane vesicles

Journal of Microbiology volume 57, pages498–508(2019)Cite this article

Abstract

Outer membrane vesicles (OMVs) are nanostructures of 20–200 nm diameter deriving from the surface of several Gram-negative bacteria. OMVs are emerging as shuttles involved in several mechanisms of communication and environmental adaptation. In this work, OMVs were isolated and characterized from Novosphingobium sp. PP1Y, a Gram-negative non-pathogenic microorganism lacking LPS on the outer membrane surface and whose genome was sequenced and annotated. Scanning electron microscopy performed on samples obtained from a culture in minimal medium highlighted the presence of PP1Y cells embedded in an extracellular matrix rich in vesicular structures. OMVs were collected from the exhausted growth medium during the mid-exponential phase, and purified by ultracentrifugation on a sucrose gradient. Atomic force microscopy, dynamic light scattering and nanoparticle tracking analysis showed that purified PP1Y OMVs had a spherical morphology with a diameter of ca. 150 nm and were homogenous in size and shape. Moreover, proteomic and fatty acid analysis of purified OMVs revealed a specific biochemical “fingerprint”, suggesting interesting details concerning their biogenesis and physiological role. Moreover, these extracellular nanostructures do not appear to be cytotoxic on HaCaT cell line, thus paving the way to their future use as novel drug delivery systems.

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Affiliations

  1. Department of Biology, University of Naples Federico II, Naples, Italy

    Federica De Lise, Francesca Mensitieri, Valeria Cafaro & Alberto Di Donato

  2. Department of Physics, University of Naples Federico II, Naples, Italy

    Giulia Rusciano & Antonio Sasso

  3. Department of Medicine, Surgery and Dentistry, University of Salerno, Fisciano, Italy

    Fabrizio Dal Piaz, Amelia Filippelli & Viviana Izzo

  4. Department of Pharmacy, University of Salerno, Fisciano, Italy

    Giovanni Forte

  5. Department of Chemical Sciences, University of Naples Federico II, Naples, Italy

    Flaviana Di Lorenzo, Antonio Molinaro, Armando Zarrelli & Valeria Romanucci

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  1. Federica De Lise
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  2. Francesca Mensitieri
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  3. Giulia Rusciano
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Correspondence to Viviana Izzo.

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De Lise, F., Mensitieri, F., Rusciano, G. et al. Novosphingobium sp. PP1Y as a novel source of outer membrane vesicles. J Microbiol. 57, 498–508 (2019). https://doi.org/10.1007/s12275-019-8483-2

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Keywords

  • outer membrane vesicles
  • exocytosis
  • bacterial secretion
  • sphingomonadales
  • Novosphingobium
  • outer cell membrane