A thermotolerant Kluyveromyces marxianus mutant was developed by exposing yeast cultures repeatedly to 48°C incubation temperature, and the strain was characterized with a significantly increased trehalose content. Unexpectedly, the strain was sensitive to alcohol, osmotic and oxidative stress, which correlated with the increases in the trehalose concentrations. Intracellular glutathione levels declined in both wild-type and mutant cells when exposed to elevating incubation temperatures. Finally, we reached the surprising conclusion that neither trehalose nor glutathione metabolisms should be aimed at in future strain development programs with K. marxianus.
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