A Physiogenomic Study of the Tolerance of <i>Saccharomyces cerevisiae</i> to Isoamyl Alcohol

Abstract

Isoamyl alcohol is a clear, unpleasantly odorous, colorless liquid of higher alcohol that emits a fruity aroma when heavily diluted. It has received much attention in recent years as a new fuel with a high energy density. Isoamyl alcohol can be produced industrially by microbial fermentation. Still, its toxicity to host cells has limited its potential for industrial production, and the molecular mechanism of its toxic effects has not yet been elucidated. In this study, RNA-Seq technology was used to analyze the transcripts of Saccharomyces cerevisiae under normal conditions and in the presence of isoamyl alcohol (0.5 g/L and 2.5 g/L). The results showed that the expression of the cell wall (CCW12, BGL2, NCW2 and SUN4), cell membrane (ELO1, ERG2, FAA1, and OPI3), translation and other structural genes were significantly down-regulated. The expression of genes related to ATP biosynthesis, NADPH biosynthesis (ZWF1), and metal ion transport (PMC1) proteins were up-regulated. Strains with key genes knocked out were cultured without isoamyl alcohol. Combined results suggested that isoamyl alcohol may affect cell wall stability and cell membrane fluidity, and the expression of genes related to ion homeostasis and energy production may play a protective role against isoamyl alcohol stress. By maintaining cell wall stability/membrane fluidity under isoamyl alcohol pressure, improving certain ion homeostasis, and generating energy/NADPH, it is possible to overcome the toxicity of isoamyl alcohol in industrial fermentation processes to a certain extent.

Keywords

• isoamyl alcohol
• <i>Saccharomyces cerevisiae</i>
• RNA-Seq
• gene knockout
• growth inhibition