Activation of the yeast Retrograde Response pathway by adaptive laboratory evolution with S-(2-aminoethyl)-L-cysteine reduces ethanol and increases glycerol during winemaking.

Publisher: BioMed Central Country of Publication: England NLM ID: 101139812 Publication Model: Electronic Cited Medium: Internet ISSN: 1475-2859 (Electronic) Linking ISSN: 14752859 NLM ISO Abbreviation: Microb Cell Fact Subsets: MEDLINE

Original Publication: London : BioMed Central, [2002-

Ethanol*/metabolism ; Wine*/analysis ; Glycerol*/metabolism ; Saccharomyces cerevisiae*/metabolism ; Saccharomyces cerevisiae*/genetics ; Fermentation* ; Saccharomyces cerevisiae Proteins*/genetics ; Saccharomyces cerevisiae Proteins*/metabolism ; Cysteine*/metabolism; Directed Molecular Evolution ; Mutation ; Acetic Acid/metabolism

Background: Global warming causes an increase in the levels of sugars in grapes and hence in ethanol after wine fermentation. Therefore, alcohol reduction is a major target in modern oenology. Deletion of the MKS1 gene, a negative regulator of the Retrograde Response pathway, in Saccharomyces cerevisiae was reported to increase glycerol and reduce ethanol and acetic acid in wine. This study aimed to obtain mutants with a phenotype similar to that of the MKS1 deletion strain by subjecting commercial S. cerevisiae wine strains to an adaptive laboratory evolution (ALE) experiment with the lysine toxic analogue S-(2-aminoethyl)-L-cysteine (AEC).
Results: In laboratory-scale wine fermentation, isolated AEC-resistant mutants overproduced glycerol and reduced acetic acid. In some cases, ethanol was also reduced. Whole-genome sequencing revealed point mutations in the Retrograde Response activator Rtg2 and in the homocitrate synthases Lys20 and Lys21. However, only mutations in Rtg2 were responsible for the overactivation of the Retrograde Response pathway and ethanol reduction during vinification. Finally, wine fermentation was scaled up in an experimental cellar for one evolved mutant to confirm laboratory-scale results, and any potential negative sensory impact was ruled out.
Conclusions: Overall, we have shown that hyperactivation of the Retrograde Response pathway by ALE with AEC is a valid approach for generating ready-to-use mutants with a desirable phenotype in winemaking.
(© 2024. The Author(s).)

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PID2021-122370OB-I00 Agencia Estatal de Investigación; PID2021-122370OB-I00 Agencia Estatal de Investigación; PID2021-122370OB-I00 Agencia Estatal de Investigación; PID2021-122370OB-I00 Agencia Estatal de Investigación

Keywords: Saccharomyces cerevisiae; Acetic acid; Adaptive laboratory evolution; Ethanol; Glycerol; Retrograde Response pathway

3K9958V90M (Ethanol)
PDC6A3C0OX (Glycerol)
0 (Saccharomyces cerevisiae Proteins)
K848JZ4886 (Cysteine)
Q40Q9N063P (Acetic Acid)

Date Created: 20240820 Date Completed: 20240821 Latest Revision: 20240823

20240823

PMC11337681

10.1186/s12934-024-02504-z

39164751