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Summary  Although mechanisms involved in response of  Saccharomyces cerevisiae  to osmotic challenge are well described for low and sudden stresses, little is known about how cells respond to a gradual increase of the osmotic pressure (reduced water activity; a w ) over several generations as it could encounter during drying in nature or in food processes. Using glycerol as a stressor, we propagated  S .  cerevisiae  through a ramp of the osmotic pressure (up to high molar concentrations to achieve testing‐to‐destruction) at the rate of 1.5 MPa day ‐1  from 1.38 to 58.5 MPa (0.990–0.635 a w ). Cultivability (measured at 1.38 MPa and at the harvest osmotic pressure) and glucose consumption compared with the corresponding sudden stress showed that yeasts were able to grow until about 10.5 MPa (0.926 a w ) and to survive until about 58.5 MPa, whereas glucose consumption occurred until 13.5 MPa (about 0.915 a w ). Nevertheless, the ramp conferred an advantage since yeasts harvested at 10.5 and 34.5 MPa (0.778 a w ) showed a greater cultivability than glycerol‐shocked cells after a subsequent shock at 200 MPa (0.234 a w ) for 2 days. FTIR analysis revealed structural changes in wall and proteins in the range 1.38–10.5 MPa, which would be likely to be involved in the resistance at extreme osmotic pressure.

Increased xerotolerance of Saccharomyces cerevisiae during an osmotic pressure ramp over several generations