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Despite the fact that Saccharomyces cerevisiae has suicide tendencies since its product affects cell function, it is a key player in alcoholic fermentation. The presence of ethanol in the medium affects membrane integrity and fluidity, as well as the rate of ethanol production. The Msn2/4p transcription factors are key regulators in stress response and play a critical role in cell response to ethanol challenge. Protein kinase A (tpk1/2/3) is controlling the activation/inactivation of a multitude of proteins through phosphorylation at specific serine residues. Targets of Protein Kinase A (PKA) are also msn2/4 and phosphorylation of these two transcription factors by PKA resulting in obstruction of their translocation to the nucleus. This work attempts to reveal the significance of specific serine residues of Msn2/4p, as possible targets of PKA, through substitution of these serine residues with alanine. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2759, 2019.

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.