Adaptive evolution of an industrial strain of Saccharomyces cerevisiae for combined tolerance to inhibitors and temperature

INTRODUCTION: Development of industrial yeast strains with high tolerance towards the inhibitors released during biomass pretreatment is critical for bioethanol production. Combining this trait with increased thermotolerance would result in a more efficient production via Simultaneous Saccharification and Fermentation (SSF) as well as reduced cooling costs. The aim of the present work was to develop a yeast strain combining these traits. RESULTS: Using a long-term adaptation strategy a stable Saccharomyces cerevisiae isolate (ISO12) was evolved from the industrial strain Ethanol Red (ER). ISO12, contrary to the parental strain, is capable of growing and fermenting the liquid fraction of non-detoxified spruce hydrolysate at 39°C with an ethanol yield of 0.38 g ethanol . g hexoses(-1). In contrast with previous studies, the superior phenotype of ISO12 does not rely on higher reductase activities for furaldehyde inhibitor conversion, but rather on a higher thermotolerance. ISO12 shows a higher capacity to ferment hydrolysate at 39°C and higher viability during heat-shock at 52°C than ER. In the absence of inhibitors, however, both ER and ISO12 displayed similar growth phenotype at 39°C. CONCLUSIONS: The evolved isolate ISO12 shows a superior phenotype than the parental strain ER when both stresses, temperature and inhibition by hydrolysate-derived compounds, are applied together. The results suggest that the presence of inhibitors depress the maximum temperature permissible for growth to a value below 39°C. As a result of the adaptation process and acquired improved thermotolerance, ISO12 is able to overcome this synergistic effect. Robust strains, such as ISO12, are interesting candidates for second generation ethanol production by SSF, as well as in tropical countries where fermentations at higher temperature can positively impact the production costs.