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Effect of ethanol perturbation on viscosity and permeability of an inner membrane in Bacillus subtilis spores

Abstract : In this work, we investigated how a combination of ethanol and high temperature (70 degrees C), affect the properties of the inner membrane of Bacillus subtilis spores. We observed membrane permeabilization for ethanol concentrations >= 50%, as indicated by the staining of the spores' DNA by the cell impermeable dye Propidium Iodide. The loss of membrane integrity was also confirmed by a decrease in the peak corresponding to dipicolinic acid using infrared spectroscopy. Finally, the spore refractivity (as measured by phase contrast microscopy) was decreased after the ethanol-heat treatment, suggesting a partial rehydration of the protoplast. Previously we have used fluorescent lifetime imaging microscopy (FLIM) combined with the fluorescent molecular rotor Bodipy-C-12 to study the microscopic viscosity in the inner membrane of B. subtilis spores, and showed that at normal conditions it is characterized by a very high viscosity. Here we demonstrate that the ethanol/high temperature treatment led to a decrease of the viscosity of the inner membrane, from 1000 cP to 860 cP for wild type spores at 50% of ethanol. Altogether, our present work confirms the deleterious effect of ethanol on the structure of B. subtilis spores, as well as demonstrates the ability of FLIM Bodipy-C-12 to measure changes in the microviscosity of the spores upon perturbation.
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Soumis le : lundi 13 février 2017 - 15:01:25
Dernière modification le : lundi 21 novembre 2022 - 03:53:24

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Pauline Loison Loison, Patrick Gervais, Jean-Marie Perrier-Cornet, Marina K. Kuimova. Effect of ethanol perturbation on viscosity and permeability of an inner membrane in Bacillus subtilis spores. Biochimica et Biophysica Acta:Biomembranes, 2016, 1858 (9), pp.2060-2069. ⟨10.1016/j.bbamem.2016.06.003⟩. ⟨hal-01466292⟩



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