Isolation of two cell populations from yeast during high-level alcoholic fermentation that resemble quiescent and nonquiescent cells from the stationary phase on glucose

High-level production of bioethanol (140 g L-1 in 45 h) in aerated fed-batch cultures of Saccharomyces cerevisiae was shown to be linked to the length of a production phase uncoupled to the growth. The induction of this phase was characterized by metabolic and morphologic changes reminiscent of those occurring in the stationary phase of growth on glucose. Global transcriptomic analysis of ethanol-stressed yeast cells in the uncoupling phase harboured features similar to those from stationary-phase cells on glucose. Two distinct cellular populations were isolated by Percoll density-gradient centrifugation in this uncoupling phase. The lower fraction was enriched by yeast cells that were mostly uniform in size and opalescent, containing a large amount of glycogen and trehalose, and exhibiting high respiratory activity. In contrast, the upper fraction was characterized by cells heterogeneous in size, with one to several small buds, which did not contain storage carbohydrates and which exhibited a poor respiratory competence while retaining a high relative glycolytic activity. These results are discussed in terms of a possible induction of a state similar to the quiescence state previously observed from yeast stationary-phase cultures, in response to ethanol toxicity, whose acquisition may be critical for performing high-level alcoholic fermentation.