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Electric field-induced effects on yeast cell wall permeabilization.

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Pulsed electric fields (PEF) increase yeast cell permeability to tetraphenylphosphonium cations (TPP+). This enhanced uptake, crucial for electroporation analysis, is controllable via PEF amplitude and duration.

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Area of Science:

  • Microbiology
  • Biophysics
  • Cell Biology

Background:

  • Yeast cell membranes present a barrier to lipophilic molecules.
  • Electroporation is a method to temporarily increase cell membrane permeability.
  • Understanding membrane permeability is key for various biotechnological applications.

Purpose of the Study:

  • To analyze the effect of pulsed electric fields (PEF) on yeast cell permeability to tetraphenylphosphonium cations (TPP+).
  • To investigate the controllability of TPP+ uptake by adjusting PEF parameters.
  • To assess the utility of TPP+ uptake measurements for studying yeast electroporation.

Main Methods:

  • Treatment of Saccharomyces cerevisiae with single square-shaped strong electric field pulses (5–150 µs, 0–10 kV/cm).
  • Analysis of TPP+ absorption using an ion-selective microelectrode (ISE).
  • Determination of plasma membrane permeability via calcein blue dye release assay.
  • Assessment of cell viability and inactivation using optical absorbance.

Main Results:

  • PEF treatment significantly increased TPP+ uptake by yeast cells after a 3-minute incubation period.
  • The extent of TPP+ uptake was directly controllable by manipulating PEF amplitude and pulse duration.
  • The kinetics of TPP+ absorption followed a second-order absolute rate equation.
  • Increased TPP+ uptake correlated with plasma membrane poration, suggesting cell wall charge changes are secondary.

Conclusions:

  • PEF effectively enhances yeast cell membrane permeability to TPP+.
  • TPP+ uptake measurements provide a viable method for analyzing the electroporation process in yeast cells.
  • The study elucidates the mechanism of PEF-induced permeability changes, involving plasma membrane poration.