Quantification of outer membrane vesicles: a potential tool to compare response in Pseudomonas putida KT2440 to stress caused by alkanols

Appl Microbiol Biotechnol. 2019 May;103(10):4193-4201. doi: 10.1007/s00253-019-09812-0. Epub 2019 Apr 10.

Abstract

The bacterial release of outer membrane vesicles (OMVs) is an important physiological mechanism of Gram-negative bacteria playing numerous key roles. One function of the release of OMVs is related to an increase in surface hydrophobicity. This phenomenon initiates biofilm formation, making bacteria more tolerant to environmental stressors. Recently, it was qualitatively shown for Pseudomonas putida that vesicle formation plays a crucial role in multiple stress responses. Yet, no quantification of OMVs for certain stress scenarios has been conducted. In this study, it is shown that the quantification of OMVs can serve as a simple and feasible tool, which allows a comparison of vesicle yields for different experimental setups, cell densities, and environmental stressors. Moreover, the obtained results provide insight to the underlying mechanism of vesicle formation as it was observed that n-alkanols, with a chain length of C7 and longer, caused a distinct and steep increase in vesiculation (12-19-fold), compared to shorter chain n-alkanols (2-4-fold increase).

Keywords: Benchmark test for stress response; Biofilm formation; Gram-negative bacteria; Outer membrane vesicles; n-alkanols.

MeSH terms

  • Alkanes / toxicity*
  • Bacterial Outer Membrane Proteins / analysis*
  • Bacteriological Techniques / methods
  • Biomarkers / analysis*
  • Extracellular Vesicles / chemistry*
  • Pseudomonas putida / drug effects*
  • Pseudomonas putida / physiology*
  • Stress, Physiological*

Substances

  • Alkanes
  • Bacterial Outer Membrane Proteins
  • Biomarkers