Ser625 of msn2 transcription factor is indispensable for ethanol tolerance and alcoholic fermentation process.

The genetic modification of yeast strains can be used as an approach for the improvement of ethanol fermentation. msn2p transcription factor is implicated in yeast stress response and its activation is controlled by protein kinase A (PKA). PKA activation inhibits the translocation of msn2p to the nucleus. An in silico analysis of msn2 protein sequence revealed serine residue at position 625 as a potent target of PKA. Thus, substitution of this serine residue with alanine increases the susceptibility of the cells to ethanol challenge reducing IC50 from 3% vol/vol to 2.42% vol/vol. Additionally, cells carrying this substitution were shown a significantly reduced fermentation rate at 30°C and 18°C increasing the total fermentation time by approximately two and three times, respectively. These results clearly indicate that Ser625 is absolutely necessary for yeast to retain its fermentation ability and ethanol tolerance.