Decreasing acetic acid accumulation by a glycerol overproducing strain of Saccharomyces cerevisiae by deleting the ALD6 aldehyde dehydrogenase gene
Abstract
Glycerol is a major fermentation product of Saccharomyces cerevisiae that contributes to the sensory character of wine. Diverting sugar to glycerol overproduction and away from ethanol production by overexpressing the glycerol 3‐phosphate dehydrogenase gene,GPD2, caused S. cerevisiae to produce more than twice as much acetic acid as the wild‐type strain (S288C background) in anaerobic cell culture. Deletion of the aldehyde dehydrogenase gene, ALD6, in wild‐type and GPD2 overexpressing strains (GPD2‐OP) decreased acetic acid production by three‐ and four‐fold, respectively. In conjunction with reduced acetic acid production, the GPD2‐OP ald6Δ strain produced more glycerol and less ethanol than the wild‐type. The growth rate and fermentation rate were similar for the modified and wild‐type strains, although the fermentation rate for the GPD2 ald6Δ strain was slightly less than that of the other strains from 24 h onwards. Analysis of the metabolome of the mutants revealed that genetic modification affected the production of some secondary metabolites of fermentation, including acids, esters, aldehydes and higher alcohols, many of which are flavour‐active in wine. Modification of GPD2 and ALD6 expression represents an effective strategy to increase the glycerol and decrease the ethanol concentration during fermentation, and alters the chemical composition of the medium such that, potentially, novel flavour diversity is possible. The implications for the use of these modifications in commercial wine production require further investigation in wine yeast strains. Copyright © 2002 John Wiley & Sons, Ltd.
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A laboratory strain of Saccharomyces cerevisiae over-expressing GPD2 and lacking ALD6 produced more glycerol and less ethanol without an increase in acetic acid.
“a laboratory strain of Saccharomyces cerevisiae over-expressing GPD2 and lacking ALD6 had the desired effect of producing more glycerol and less ethanol, without an increase in acetic acid (Remize et al. 2000, Eglinton et al. 2002).”
From Yeast and bacterial modulation of wine aroma and flavour (2005) · cites this paper as Eglinton et al. 2002
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MethodUpregulation ofALD3andGPD1inSaccharomyces cerevisiaeduring Icewine fermentation
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Quantitative trait analysis of yeast biodiversity yields novel gene tools for metabolic engineering
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“side-effect of higher acetate production (Cambon et al., 2006; Eglinton et al., 2002; Ehsani et al., 2009).”
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“It has previously been reported that Ald6p is the main aldehyde dehydrogenase responsible for acetic acid production in Saccharomyces cerevisiae strains during fermentation of glucose media (Eglinton et al. 2002), although most recently, mitochondrial Ald5p was also found to contribute (Saint-Prix et al.”
BackgroundIs there a future in glycerol as a feedstock in the production of biofuels and biochemicals?
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“The problem, however, was alleviated quite simply by knocking out ALD6.”
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“This involves conversion of pyruvate to acetaldehyde by pyruvate decarboxylase, which is in turn oxidized to acetate by aldehyde dehydrogenase (ALD).”
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“By contrast, the fluxes through glycerol synthesis [54–57] or the PPP [42, 44] were more flexible, which makes them more interesting targets to redirect metabolic fluxes.”
BackgroundKim, S. R., Skerker, J. M., Kong, I. I., Kim, H., Maurer, M. J., Zhang, G.-C., Peng, D., Wei, N., Arkin, A. P., & Jin, Y. (2017) · Metabolic Engineering · doi:10.1016/j.ymben.2017.02.006 · Cited by 40 · see citing works
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Oenological Characterization of Native Hanseniaspora uvarum Strains
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“exposed to sugar media or juice (Eglinton et al. 2002; Erasmus et al. 2003; Erasmus and van Vuuren 2009; Noti et al. 2015), although both mitochondrial isoforms (NADP-dependent Ald4p and NAD/NADP-dependent Ald5p) were also found to contribute (Remize et al.”
BackgroundPre-fermentative supplementation of fatty acids alters the metabolic activity of wine yeasts
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“In addition, acetic acid can also be formed by the metabolism of heterofermentative LAB and wild yeasts (Eglinton et al. 2002; Endo and Dicks 2014).”
BackgroundWine Yeasts Selection: Laboratory Characterization and Protocol Review
Sidari, R., Ženišová, K., Tobolková, B., Belajová, E., Cabicarová, T., Bučková, M., Puškárová, A., Planý, M., Kuchta, T., & Pangallo, D. (2021) · Microorganisms · doi:10.3390/microorganisms9112223 · Cited by 32 · see citing works
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“However, deletion of the aldehyde dehydrogenase gene, ALD6, in wild-type and GPD2 overexpressing strains decreased acetic acid production by three- and fourfold, respectively (Eglinton et al. 2002).”
BackgroundEffects ofADH2Overexpression inSaccharomyces bayanusduring Alcoholic Fermentation
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“Genetic engineering strategies aimed at modifying glycerol accumulation in wines have exposed the difficulty of altering yeast metabolism in the desired direction without introducing unwanted side effects (15, 26, 32).”
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Torulaspora delbrueckii produces high levels of C5 and C6 polyols during wine fermentations
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“Yeast is also an attractive host for the production of propionic acid (PA) that is commonly used as a food preservative and a chemical intermediate, since PA can be formed as a by‐product of yeast fermentation (Eglinton et al., 2002).”
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Bio-Dealcoholization of Wines: Can Yeast Make Lighter Wines?
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“Ald aldehyde dehydrogenase enzymes later help to restore the yeast’s redox balance by reducing coenzymes NAD + or NADP + during the oxidation of acet-aldehyde to acetic acid and acetoin (Eglinton et al. 2002).”
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“The microbiological approach includes the use of non-GMO (genetically modified organisms) strategies like sequential inoculations and co-inoculations of S. cerevisiae with non-Saccharomyces yeasts, as well as GMO-based strategies with the use of genetically modified yeasts.”
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Yeast selection for wine flavour modulation
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- Authors
- Eglinton, J. M., Heinrich, A. J., Pollnitz, A. P., Langridge, P., Henschke, P. A., & de Barros Lopes, M.
- Affiliations
- Australian Wine Research Institute (AU)
- The University of Adelaide (AU)
- Corresponding author
- Eglinton, Jeffrey M.
- ORCID
- Langridge, Peter
- Venue
- Yeast
- Publisher
- Wiley
- Published
- 2002-01-18
- Volume
- 19
- Issue
- 4
- Pages
- 295-301
- ISSN
- 0749-503X (ISSN-L)
- 1097-0061
- OpenAlex
- W2080704515
- Cited by
- 160
- References
- 23
- Citations by year
- 2026: 1 · 2025: 5 · 2024: 14 · 2023: 2 · 2022: 4 · 2021: 6 · 2020: 8 · 2019: 8 · 2018: 5 · 2017: 10 · 2016: 10 · 2015: 5 · 2014: 4 · 2013: 4 · 2012: 11
- FWCI
- 10.74
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Yeast (publishedVersion)
- PubMed (publishedVersion)
- Adelaide Research & Scholarship (AR&S) (University of Adelaide) (submittedVersion)
- Topics
- Life Sciences › Agricultural and Biological Sciences › Food Science › Fermentation and Sensory Analysis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Keywords
- Saccharomyces cerevisiae, acetate accumulation, aldehyde dehydrogenase, glycerol-3-phosphate dehydrogenase, wine yeast, wine flavor, ethanol production, secondary metabolites, anaerobic fermentation
- MeSH terms
- Aldehyde Dehydrogenase, Fermentation, Glycerol, Glycerolphosphate Dehydrogenase, Saccharomyces cerevisiae, Gene Deletion, Acetic Acid
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