Abstract
Acid stress is one of the most common adverse conditions during fermentation of fruit wines, and the acid tolerance of yeasts is, therefore, critical for fruit wine production. However, the biological mechanism underlying the acquired tolerance of yeasts against acid stress is poorly understood. We have previously obtained an evolved Saccharomyces cerevisiae strain ET008-c54 with increased tolerance against acid stress, and potentially, it serves as a promising yeast strain for greengage wine fermentation. In the current study, we further revealed the alterations responsible for the adaptation of ET008-c54 to low pH by whole-genome re-sequencing, transcriptomic, and metabolic analyses. Results confirmed the outstanding fermenting performance of ET008-c54 at low pH as compared with the parental ET008. More specifically, the growth rate of ET008-c54 at low pH was increased by 6.24 times and the fermentation time was shortened by 70%. Differences were also observed in the physiology of the strains through ergosterol, H+-ATPase activity, and aroma determinations. By integrating both RNA-seq and whole-genome re-sequencing data, we demonstrated some metabolic pathways in ET008-c54, namely ergosterol synthesis and ferrous iron uptake, in which several acid-responsive genes were involved being upregulated. Also, upregulation of the pathways responsible for aroma compound formation, including fatty acid ethyl ester synthesis and aromatic amino acid biosynthesis, was identified. Thus, the enhanced fermentation ability of ET008-c54 at low pH should be, at least partly, contributed by the altered gene expressions associated with the aforementioned pathways. By elucidating the biological mechanism of yeasts against acid stress, this current study allows better-defined targets for future studies of genetic improvement of wine yeasts and enhancement of the fermentation processes.
Key points
• Metabolic analysis confirmed the excellent fermentation performance of ET008-c54.
• Acid tolerance genes for ergosterol synthesis and ferrous iron uptake were upregulated.
• Aroma genes for fatty acid ethyl ester and aromatic amino acid synthesis were upregulated.
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Funding
This work was supported by the National Natural Science Foundation of China (grant number 31701588), the Natural Science Foundation of Jiangsu Province (grant number BK20170178), the Fundamental Research Funds for the Central Universities (grant number JUSRP11965 and JUSRP21914), and the Program of Introducing Talents of Discipline to Universities (111 Project) (111-2-06).
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TT, DW, and JL conceived and designed research. TT, HY, and JL conducted experiments. TT and JS analyzed data. TT and CN wrote the manuscript. All authors read and approved the manuscript.
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Tian, T., Wu, D., Ng, CT. et al. Uncovering mechanisms of greengage wine fermentation against acidic stress via genomic, transcriptomic, and metabolic analyses of Saccharomyces cerevisiae. Appl Microbiol Biotechnol 104, 7619–7629 (2020). https://doi.org/10.1007/s00253-020-10772-z
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DOI: https://doi.org/10.1007/s00253-020-10772-z