Osmoporation: a simple way to internalize hydrophilic molecules into yeast

Abstract

Internalization of hydrophilic molecules into yeast cytosol is required for different applications such as cell transformation or preservation of water soluble components by bioencapsulation. However, these molecules are not able to cross the plasma membrane and strategies have to be developed. Recent works revealed that osmotic perturbations could induce non-lethal transient permeabilization of the plasma membrane. In this work, we endeavored to clarify the phenomenon of permeabilization during rehydration after a mild hyperosmotic perturbation in order to evaluate the possibility of hydrophilic molecule internalization in yeast by this treatment. Rehydration step is particularly interesting because the large entry of water into the cells could help the internalization of molecules. The internalization of a fluorescent molecule [fluorescein isothiocyanate Dextran (FITC-Dextran), 20 kDa], added during the rehydration after a sublethal hyperosmotic treatment, was studied in Saccharomyces cerevisiae yeast cells. The internalization kinetic and the localization of the fluorescent molecules were studied by flow cytometry and fluorescence confocal microscopy. Our results show that the rehydration leads to the rapid internalization of FITC-Dextran due to a transient plasma membrane permeabilization. Thus, osmoporation, i.e. plasma membrane poration by modifications of osmotic pressure of the extracellular medium, could be a new and simple way to deliver molecules of particular interest into yeasts.

Curcumin and fisetin internalization into Saccharomyces cerevisiae cells via osmoporation: impact of multiple osmotic treatments on the process efficiency

“…Glycerol solutions were prepared to the final osmotic pressures ( π ) of 1·4 MPa ( iso ‐osmotic) and 30·0 MPa, respectively, according to da Silva Pedrini et al . ().…”

“…Despite the high EE, the loading amount of nanocapsules' internal space represented only 2Á8% of capsules. Previous studies (da Silva Pedrini et al 2014;Câmara Junior et al 2016) have shown that osmoporation is an efficient technique regarding the use of available space in the yeast cells, as the encapsulated molecule is able to occupy and diffuse throughout the space of cell's cytoplasm.…”

“…Food-grade S. cerevisiae (Fleischmann, Jundia ı, Brazil) was cultivated according to the conditions described by da Silva Pedrini et al (2014). At harvesting conditions (cell density approximately 10 8 cells per ml), 40 ml of the cell suspension was centrifuged at 2200 g at 25°C for 5 min (Solab, Piracicaba, Brazil).…”

“…We demonstrated that the osmotic treatment promoted a more efficient internalization when compared to natural endocytosis and that encapsulated content was dispersed throughout cell cytoplasm (da Silva Pedrini et al . ). Furthermore, we also reported that using suitable process conditions, hydrophobic low molecular weight bioactive molecules (fisetin) could also be incorporated into yeast cells via osmoporation (Câmara Junior et al .…”

Can biological structures be natural and sustainable capsules?

“…Therefore, there are several strategies between passive transfer and solvent-based “cleaning” of the shell of the structure as it is done for instance for pollen hollow shell (Diego-Taboada et al, 2014 ). A treatment of osmoporation has also been proposed to increase the transfer (da Silva Pedrini et al, 2014 ) showing an interest for the highly hydrosoluble chlorogenic acid (Shi et al, 2007 ) but not for the less polar curcumin (Paramera et al, 2011 ).…”