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Indentation with atomic force microscope,Saccharomyces cerevisiae cell gains elasticity under ethanol stress
Institution:1. Departamento de Biotecnología, Instituto de Agroquímica y Tecnología de los Alimentos, CSIC, Carrer del Catedràtic Agustín Escardino Benlloch, 7, 46980 Paterna, Valencia, Spain;2. Instituto de Investigación y Desarrollo en Ingeniería de Procesos, Biotecnología y Energías Alternativas, PROBIEN, Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina, Universidad Nacional del Comahue, Buenos Aires 1400, 8300, Neuquén, Argentina;3. Facultad de Ciencias Médicas, Universidad Nacional del Comahue, 8324 Cipolletti, Río Negro, Argentina;4. Facultad de Ciencias Agrarias, Universidad Nacional del Comahue, 8303 Cinco Saltos, Río Negro, Argentina
Abstract:During bioethanol fermentation process, Saccharomyces cerevisiae cell membrane is the first target to be attacked by the accumulated ethanol. In such a prominent position, S. cerevisiae cell membrane could reversely provide protection through changing fluidity or elasticity secondary to remodeled membrane components or structure during the fermentation process. However, there is yet to be a direct observation of the real effect of the membrane compositional change. In this study, atomic force microscope-based strategy was performed to determine Young's modulus of S. cerevisiae to directly clarify ethanol stress-associated changes and roles of S. cerevisiae cell membrane fluidity and elasticity. Cell survival rate decreased while the cell swelling rate and membrane permeability increased as ethanol concentration increased from 0% to 20% v/v. Young's modulus decreased continuously from 3.76 MPa to 1.53 MPa while ethanol stress increased from 0% to 20% v/v, indicating that ethanol stress induced the S. cerevisiae membrane fluidity and elasticity changes. Combined with the fact that membrane composition varies under ethanol stress, to some extent, this could be considered as a forced defensive act to the ethanol stress by S. cerevisiae cells. On the other hand, the ethanol stress induced loosening of cell membrane also caused S. cerevisiae cell to proactively remodel membrane to make cell membrane more agreeable to the increase of environmental threat. Increased ethanol stress made S. cerevisiae cell membrane more fluidized and elastic, and eventually further facilitated yeast cell’s survival.
Keywords:Ethanol tolerance  Membrane fluidity and elasticity  Young's modulus  Atomic force microscope
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