Oxygen Response of the Wine Yeast Saccharomyces cerevisiae EC1118 Grown under Carbon-Sufficient, Nitrogen-Limited Enological Conditions
Felipe F. AceitunoMarcelo OrellanaJorge TorresSebastián N. MendozaAlex W. SlaterFrancisco MeloEduardo Agosín
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Discrete additions of oxygen play a critical role in alcoholic fermentation. However, few studies have quantitated the fate of dissolved oxygen and its impact on wine yeast cell physiology under enological conditions. We simulated the range of dissolved oxygen concentrations that occur after a pump-over during the winemaking process by sparging nitrogen-limited continuous cultures with oxygen-nitrogen gaseous mixtures. When the dissolved oxygen concentration increased from 1.2 to 2.7 μM, yeast cells changed from a fully fermentative to a mixed respirofermentative metabolism. This transition is characterized by a switch in the operation of the tricarboxylic acid cycle (TCA) and an activation of NADH shuttling from the cytosol to mitochondria. Nevertheless, fermentative ethanol production remained the major cytosolic NADH sink under all oxygen conditions, suggesting that the limitation of mitochondrial NADH reoxidation is the major cause of the Crabtree effect. This is reinforced by the induction of several key respiratory genes by oxygen, despite the high sugar concentration, indicating that oxygen overrides glucose repression. Genes associated with other processes, such as proline uptake, cell wall remodeling, and oxidative stress, were also significantly affected by oxygen. The results of this study indicate that respiration is responsible for a substantial part of the oxygen response in yeast cells during alcoholic fermentation. This information will facilitate the development of temporal oxygen addition strategies to optimize yeast performance in industrial fermentations.Keywords:
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Remarks Concerning the Expression of Certain Parameters of Must and Wine Composition. Preface to the First Edition. Preface to the Second Edition. 1. Cytology, Taxonomy and Ecology of Grape and Wine Yeasts. 2. Biochemistry of Alcoholic Fermentation and Metabolic Pathways of Wine Yeasts. 3. Conditions of Yeast Development. 4. Lactic Acid Bacteria. 5. Metabolism of Lactic Acid Bacteria. 6. Lactic Acid Bacteria Development in Wine. 7. Acetic Acid Bacteria. 8. The Use of Sulfur Dioxide in Must and Wine Treatment. 9. Products and Methods Complementing the Effect of Sulfur Dioxide. 10. The Grape and its Maturation. 11. Harvest and Pre-Fermentation Treatments. 12. Red Winemaking. 13. White Winemaking. 14. Other Winemaking Methods. Index.
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Abstract BACKGROUND Malolactic fermentation ( MLF ) is a critical step in modern winemaking and the co‐inoculation of lactic acid bacteria with yeasts represents an emerging approach to improve the quality of wine. This study aims to evaluate the effect of inoculation timing on the chemical and sensory characteristics of two commercial Cabernet Franc wines: (i) a control wine in which MLF was inoculated after the completion of alcoholic fermentation ( AF ); and (ii) a co‐inoculated wine where simultaneous alcoholic and malolactic fermentations occurred. RESULTS Besides the expected full conversion of malic acid into lactic acid, the MLF with co‐inoculum of yeast/bacteria at initial AF allowed for reduced fermentation times, i.e. faster winemaking process. Although important changes in the chemical parameters of wines were found, especially color intensity and volatile compounds, there was a trend on the sensory evaluation of wines with a greater perception of red and ripe fruits in the case of co‐inoculation wine, while the control wine was dominated by notes of spice and herbs. The results of GC / MS analysis showed the largest differences for the 3‐hydroxy‐2‐butanone (acetoin, the reduced form of diacetyl) and isoamyl alcohol concentrations, the former was 3.2 times higher under co‐inoculum conditions, whereas the latter showed the opposite behavior. CONCLUSION The co‐inoculation protocol proposed is a valuable choice for innovative winemaking of Cabernet Franc wines, and results from a large‐scale winemaking process are provided for the first time. © 2015 Society of Chemical Industry
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The grape used in wine making has many wild microorganisms like lactic acid bacteria, yeast, acetic acid bacteria. During the alcoholic fermentation, the evaluation of these microorganisms depends on their activity. There is an interaction between yeast and lactic acid bacteria during this period of wine making. In this study, we have made wine from the autochthonous Albanian grape Kallmet variety using the spontaneous fermentation and inoculated fermentation with the yeast Saccharomyces bayannus. Yeasts carry out the alcohol fermentation, and lactic acid bacteria make malolactic fermentation in wine. With this fermentation, lactic acid bacteria convert malic acid to lactic acid, reducing the acidity of the wine and create a microbiological stability. During the alcoholic fermentation, the evaluation of lactic acid bacteria is not required. The aim of our study is to evaluate the first quantity of lactic acid bacteria to Kallmet grape, their performance during the two fermentations, spontaneous and inoculated fermentations.
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Yeasts are predominant in the ancient and complex process of winemaking. In spontaneous fermentations, there is a progressive growth pattern of indigenous yeasts, with the final stages invariably being dominated by the alcohol-tolerant strains of Saccharomyces cerevisiae. This species is universally known as the 'wine yeast' and is widely preferred for initiating wine fermentations. The primary role of wine yeast is to catalyze the rapid, complete and efficient conversion of grape sugars to ethanol, carbon dioxide and other minor, but important, metabolites without the development of off-flavours. However, due to the demanding nature of modern winemaking practices and sophisticated wine markets, there is an ever-growing quest for specialized wine yeast strains possessing a wide range of optimized, improved or novel oenological properties. This review highlights the wealth of untapped indigenous yeasts with oenological potential, the complexity of wine yeasts' genetic features and the genetic techniques often used in strain development. The current status of genetically improved wine yeasts and potential targets for further strain development are outlined. In light of the limited knowledge of industrial wine yeasts' complex genomes and the daunting challenges to comply with strict statutory regulations and consumer demands regarding the future use of genetically modified strains, this review cautions against unrealistic expectations over the short term. However, the staggering potential advantages of improved wine yeasts to both the winemaker and consumer in the third millennium are pointed out.
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Wine is the product of complex interactions between yeasts and bacteria in grape must. Amongst yeast populations, two groups can be distinguished. The first, named non-Saccharomyces (NS), colonizes, with many other micro-organisms, the surface of grape berries. In the past, NS yeasts were primarily considered as spoilage micro-organisms. However, recent studies have established a positive contribution of certain NS yeasts to wine quality. Amongst the group of NS yeasts, Brettanomyces bruxellensis, which is not prevalent on wine grapes, plays an important part in the evolution of wine aroma. Some of their secondary metabolites, namely volatile phenols, are responsible for wine spoilage. The other group contributing to wine aroma, which is also the main agent of alcoholic fermentation (AF), is composed of Saccharomyces species. The fermenting must is a complex microbial ecosystem where numerous yeast strains grow and die according to their adaptation to the medium. Yeast-yeast interactions occur during winemaking right from the onset of AF. The aim of this study was to describe the interactions between B. bruxellensis, other NS and Saccharomyces cerevisiae during laboratory and practical scale winemaking.Molecular methods such as internal transcribed spacer-restriction fragment length polymorphism and polymerase chain reaction and denaturing gradient gel electrophoresis were used in laboratory scale experiments and cellar observations. The influence of different oenological practices, like the level of sulphiting at harvest time, cold maceration preceding AF, addition of commercial active dry yeasts on B. bruxellensis and other yeast interactions and their evolution during the initial stages of winemaking have been studied. Brettanomyces bruxellensis was the most adapted NS yeast at the beginning of AF, and towards the end of AF it appeared to be more resistant than S. cerevisiae to the conditions of increased alcohol and sugar limitation.Among all NS yeast species, B. bruxellensis is better adapted than other wild yeasts to resist in must and during AF. Moreover, B. bruxellensis appeared to be more tolerant to ethanol stress than S. cerevisiae and after AF B. bruxellensis was the main yeast species in wine.Brettanomyces bruxellensis interacts with other yeast species and adapts to the wine medium as the dominant yeast species at the end of AF. Contamination of B. bruxellensis might take place at the beginning of malolactic fermentation, which is a critical stage in winemaking.
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