Emilia Matallana

University of Valencia | UV · Biochemistry and Molecular Biology

The bulk of grape juice fermentation is carried out by the yeast Saccharomyces cerevisiae, but non-Saccharomyces yeasts can modulate many sensorial aspects of the final products in ways not well understood. In this study, some of such non-conventional yeasts were screened as mixed starter cultures in a defined growth medium in both simultaneous and...

The bulk of grape juice fermentation is carried out by the yeast Saccharomyces cerevisiae , but non- Saccharomyces yeasts can modulate many sensorial aspects of the final products in ways not well understood. In this study, some of such non-conventional yeasts were screened as mixed starter cultures in a fermentation defined medium in both simultan...

Wine strains of Saccharomyces cerevisiae have to adapt their metabolism to the changing conditions during their biotechnological use, from the aerobic growth in sucrose-rich molasses for biomass propagation to the anaerobic fermentation of monosaccharides of grape juice during winemaking. Yeast have molecular mechanisms that favor the use of prefer...

Modern enology relies on the use of selected yeasts, both Saccharomyces and non-conventional, as starters to achieve reliable fermentations. That allows the selection of the right strain for each process and also the improvement of such strain, by traditional methods or approaches involving genetic manipulation. Genetic engineering allows deletion,...

Peroxiredoxins are a family of peroxide-degrading enzymes for challenging oxidative stress. They receive their reducing power from redox-controlling proteins called thioredoxins, and these, in turn, from thioredoxin reductase. The main cytosolic peroxiredoxin is Tsa1, a moonlighting protein that also acts as protein chaperone a redox switch control...

The potential of yeasts isolated from traditional chichas as starter cultures, either for controlled production of the native beverage or for industrial beer production, has been investigated. Three S. cerevisiae strains and one T. delbrueckii strain isolated from four different Ecuadorian chichas were compared to ale and lager beer strains with re...

The ability of the yeast Saccharomyces cerevisiae to adapt to the changing environment of industrial processes lies in the activation and coordination of many molecular pathways. The most relevant ones are nutrient signaling pathways because they control growth and stress response mechanisms as a result of nutrient availability or scarcity and, the...

Background: Saccharomyces cerevisiae wine strains can develop stuck or sluggish fermentations when nutrients are scarce or suboptimal. Nutrient sensing and signaling pathways, such as PKA, TORC1 and Snf1, work coordinately to adapt growth and metabolism to the amount and balance of the different nutrients in the medium. This has been exhaustively...

Non-Saccharomyces wine yeasts are useful tools for producing wines with complex aromas or low ethanol content. Their use in wine would benefit from their production as active dry yeast (ADY) starters to be used as co-inocula alongside S. cerevisiae. Oxidative stress during biomass propagation and dehydration is a key factor in determining ADY perfo...

Yeasts involved in the spontaneous fermentation of traditional beverages like chicha (indigenous Andean beer) may have the potential to be used as starter cultures to improve the quality and microbiological safety of these products, but also as non-conventional alternatives to other food alcoholic fermentations. In this research, we isolated, ident...

Yeasts are very important microorganisms for food production. The high fermentative capacity, mainly of the species of the genus Saccharomyces, is a key factor for their biotechnological use, particularly to produce alcoholic beverages. As viability and vitality are essential to ensure their correct performance in industry, this review addresses th...

Oxidative stress is a common hazardous condition that cells have to face in their lifetime. Oxidative damage may diminish cell vitality and viability by reducing metabolism and eventually leading to aging and ultimate death. Wine yeast Saccharomyces cerevisiae also faces oxidative attack during its biotechnological uses. One of the main yeast antio...

The thioredoxin system plays a predominant role in the control of cellular redox status. Thioredoxin reductase fuels the system with reducing power in the form of NADPH. The TORC1 complex promotes growth and protein synthesis when nutrients, particularly amino acids, are abundant. It also represses catabolic processes, like autophagy, which are act...

Biomass propagation for the production of active dry yeasts (ADY) is an economically important industrial process where cellular oxidative stress significantly limits yield and fermentative capacity in the final product. Oxidative stress affects macromolecular cell components, such as lipid and proteins, thus impairing many different cellular proce...

Several yeast species, belonging to Saccharomyces and non-Saccharomyces genera, play fundamental roles during spontaneous must grape fermentation, and recent studies have shown that mixed fermentations, co-inoculated with S. cerevisiae and non-Saccharomyces strains, can improve wine organoleptic properties. During active dry yeast (ADY) production,...

Glufosinate ammonium (GA) is a widely used herbicide that inhibits glutamine synthetase. This inhibition leads to internal amino acid starvation which, in turn, causes the activation of different nutrient sensing pathways. GA also inhibits the enzyme of the yeast Saccharomyces cerevisiae in such a way that, although it is not used as a fungicide, i...

We provide an optimized protocol for a double staining technique to analyze superoxide dismutase enzymatic isoforms Cu-Zn SOD (Sod1) and Mn-SOD (Sod2) and catalase in the same polyacrylamide gel. The use of NaCN, which specifically inhibits yeast Sod1 isoform, allows the analysis of Sod2 isoform while the use of H2O2 allows the analysis of catalase...

Grape juice fermentation is a harsh environment with many stressful conditions, and Saccharomyces cerevisiae adapts its metabolism in response to those environmental challenges. Many nutrient-sensing pathways control this feature. The Tor/Sch 9p pathway promotes growth and protein synthesis when nutrients are plenty, while the transcription factor...

Wine yeast deals with many stress conditions during its biotechnological use. Biomass production and its dehydration produce major oxidative stress, while hyperosmotic shock, ethanol toxicity and starvation are relevant during grape juice fermentation. Most stress response mechanisms described in laboratory strains of Saccharomyces cerevisiae are u...

Glycerol is a key yeast metabolite in winemaking because it contributes to improve the organoleptic properties of wine. It is also a cellular protective molecule that enhances the tolerance of yeasts to osmotic stress and promotes longevity. Thus, its production increases by genetic manipulation, which is of biotechnological and basic interest. Gly...

The tolerance of the yeast Saccharomyces cerevisiae to desiccation is important for the use of this microorganism in the wine industry, since active dry yeast (ADY) is routinely used as starter for must fermentations. Both biomass propagation and dehydration cause cellular oxidative stress, therefore negatively affecting yeast performance. Protecti...

Saccharomyces cerevisiae chronological life span (CLS) is determined by a wide variety of environmental and genetic factors. Nutrient limitation without malnutrition, i.e. dietary restriction, expands CLS through the control of nutrient signaling pathways, of which TOR/Sch9 has proven to be the most relevant, particularly under nitrogen deprivation...

Mitochondria are the cell's powerhouse when organisms are grown in the presence of oxygen. They are also the source of reactive oxygen species that cause damage to the biochemical components of the cell and lead to cellular aging and death. Under winemaking conditions, Saccharomyces yeasts exclusively have a fermentative metabolism due to the high...

During yeast biomass production, cells are grown through several batch and fed-batch cultures on molasses. This industrial process produces several types of stresses along the process, including thermic, osmotic, starvation and oxidative stress. It has been shown that Saccharomyces cerevisiae strains with enhanced stress resistance present enhanced...

The production of active dried yeast (ADY) is a common practice in industry to maintain yeast starters and facilitate its storage for long periods, however, it causes multiple cell injuries, oxidative damage being one of the most important stresses. Dehydration tolerance is then a highly appreciated property in yeast for ADY production. In the pres...

Cell-to-cell and cell-to-environment interactions of microorganisms are of substantial relevance for their biotechnological use. In the yeast Saccharomyces cerevisiae, flocculation can be an advantage to clarify final liquid products after fermentation, and biofilm formation may be relevant for the encapsulation of strains of interest. The adhesion...

The whole genome analysis has demonstrated that wine yeasts undergo changes in promoter regions and variations in gene copy number, which make them different to lab strains and help them better adapt to stressful conditions during winemaking, where oxidative stress plays a critical role. Since cytoplasmic thioredoxin II, a small protein with thiol-...

Loss of function mutations in the DJ-1 gene are linked to rare autosomal recessive forms of Parkinsonism. In Drosophila, two DJ-1 orthologs have been identified, DJ-1α and DJ-1β. Several studies have shown that DJ-1β mutant flies are viable and fertile but exhibit age-dependent locomotor defects, shortened lifespan as well as enhanced sensitivity t...

Aerobic organisms have devised several enzymatic and non-enzymatic antioxidant defenses to deal with reactive oxygen species (ROS) produced by cellular metabolism. To combat such stress, cells induce ROS scavenging enzymes such as catalase, peroxidase, superoxide dismutase (SOD) and glutathione reductase. In the present research, we have used a dou...

Background Yeast viability and vitality are essential for different industrial processes where the yeast Saccharomyces cerevisiae is used as a biotechnological tool. Therefore, the decline of yeast biological functions during aging may compromise their successful biotechnological use. Life span is controlled by a variety of molecular mechanisms, ma...

Sugar consumption during grape juice fermentation by mutants in mRNA binding proteins (A) and apoptosis –related genes (B). The data reflect the experiments shown in Figure 4B and Figure 7A respectively.

Yeast strains used in this work.

Oligonucleotides used in this work.

Fermentation in synthetic grape juice of HST3 overproducing strains (A) Dry biomass production. (B) Ethanol production. *p<0.05, ***p<0.005, unpaired t-test, two-tailed.

Background Viability in a non dividing state is referred to as chronological life span (CLS). Most grape juice fermentation happens when Saccharomyces cerevisiae yeast cells have stopped dividing; therefore, CLS is an important factor toward winemaking success. Results We have studied both the physical and chemical determinants influencing yeast C...

Figure S1. CLS analysis of the EC1118 strain in water containing ethanol (8 g/L), acetate (0.4 g/L) and acetaldehyde (0.12 g/L). The assays were performed as described in Figure 1. Experiments were done in triplicate, and the mean and standard deviation are provided.

Figure S2. Production of ethanol (A), acetaldehyde (B) and acetic acid (C) during the grape juice fermentation in synthetic grape juice containing 2 mg/L resveratrol or 9 mg/L quercetin described in Figure 2A. Experiments were done in triplicate, and the mean and standard deviation are provided.

Table S1. Initial viability of the chronological life span experiments (in cfu/mL x106)

Grape juice fermentation by wine yeast is an interesting model to understand aging under conditions closer to those in nature. Grape juice is rich in sugars and, unlike laboratory conditions, the limiting factor for yeast growth is nitrogen. We tested the effect of deleting sirtuins and several acetyltransferases to find that the role of many of th...

ABSTRACT: Following publication of this work [Gomez-Pastor et al, Microbial Cell Factories 2010, 9:9] we have noticed a production error in the article. Figure 1 in the original version showed incorrect results, with graphs having been duplicated in error from another figure. The correct results for Figure 1 are shown below. Legend to Figure 1 Impr...

Most grape juice fermentation takes place when yeast cells are in a nondividing state called the stationary phase. Under such circumstances, we aimed to identify the genetic determinants controlling longevity, known as the chronological life span. We identified commercial strains with both short (EC1118) and long (CSM) life spans in laboratory grow...

In the yeast biomass production process, protein carbonylation has severe adverse effects since it diminishes biomass yield and profitability of industrial production plants. However, this significant detriment of yeast performance can be alleviated by increasing thioredoxins levels. Thioredoxins are important antioxidant defenses implicated in man...

Carbonylation profile of the different protein functional categories during biomass propagation process. Relative carbonyl content measured as CI/PI of each defined functional category at 15 h and 80 h of growth among the three strains T73, TTRX2 and trx2. All the data are expressed as means ± standard deviation. Comparisons among multiple groups w...

Carbonyls levels after drying process and YPGF re-inocculation. (A) anti-DNP monodimensional western blot at different time points of the industrial process and after drying and re-inocculation in YPGF for T73 and TTRX2 strains. (B) Relative carbonyl levels were quantified by using the QuantityOne software (Bio-Rad) and carbonylation levels were no...

Trx2p visualization during biomass propagation. (A). Magnified regions of the two-dimensional gels where Trx2p can be observed in the different strains (A) T73 and (B) TTRX2 at 15 h of the biomass propagation process. (B) Western blot anti-Trx2p during biomass propagation. Coomassie-stained membranes are shown as a control of protein amount.

In the industrial yeast biomass production process, cells undergo an oxidative and other stresses which worsen the quality of the produced biomass. The overexpression of the thioredoxin codifying gene TRX2 in a wine Saccharomyces cerevisiae strain increases resistance to oxidative stress and industrial biomass production yield. We observed that var...

This chapter discusses the microbiological aspects of saccharomyces yeasts in wine production. Yeasts, which play a central role in the winemaking process, are unicellular fungi that reproduce by budding. Most yeast belongs to the phylum Ascomycota on the basis of their sexual development. The selection of wine yeasts with specific genetic markers...

Transcriptome and proteome profiles have been established for the commercial wine yeast strain T73 during an important industrial process: yeast biomass propagation. The data from both analyses reveal that the metabolic transition from fermentation to respiration is the most critical step in biomass propagation. We identified 177 ORFs and 56 protei...

The beneficial effect of improving yeast redox response by increasing thioredoxin levels has been shown. Decreased lipid and protein oxidation is reflected in an increased biomass yield. In addition, increased redox defenses like glutathione and ROS scavenging enzymes are observed. Furthermore, the wine produced with the modified strain presented m...

Mutations in the DJ-1 gene cause autosomal recessive, early-onset Parkinsonism. The DJ-1 protein exerts a protective role against oxidative stress damage, working as a cellular oxidative stress sensor, and it seems to regulate gene expression at different levels. In Drosophila, two DJ-1 orthologs have been identified: DJ-1β and DJ-1β. Several studi...

Growth parameters for the improved TTRX2 strain compared to control T73 and T73Yep352 strains. Growth analysis for T73 (black circle), TTRX2 (black triangle) and T73Yep352 (white square) strains on YPD plates (A) or liquid (B) medium. No significant differences were observed. Plates were spotted with 5 μl of exponential cultures equaled to OD = 0.1...

Protein carbonylation along bench-top trials of biomass propagation for T73 and TGSH1 strain. Western analysis of oxidatively damaged proteins (top panels) and total protein stain (bottom panels) for T73 (left panels) and TGSH1 (right panels) strains (A). Panel B shows quantification of protein carbonyl content of T73 (black circles) and TGSH1 (ope...

Wine Saccharomyces cerevisiae strains, adapted to anaerobic must fermentations, suffer oxidative stress when they are grown under aerobic conditions for biomass propagation in the industrial process of active dry yeast production. Oxidative metabolism of sugars favors high biomass yields but also causes increased oxidation damage of cell components...

The tolerance of the yeast Saccharomyces cerevisiae to desiccation is important for the use of this microorganism in the wine industry, since active dry wine yeast is routinely used as starter for must fermentations. Many studies have shown the complexity of the cellular effects caused by water loss, including oxidative injuries on macromolecular c...

Trehalose accumulation is a common response to several stresses in the yeast Saccharomyces cerevisiae. This metabolite protects proteins and membrane lipids from structural damage and helps cells to maintain integrity. Based on genetic studies, degradation of trehalose has been proposed as a required mechanism for growth recovery after stress, and...

The role of the acid trehalase encoded by the ATH1 gene in the yeast Saccharomyces cerevisiae is still unclear. In this work, we investigated the regulation of ATH1 transcription and found a clear involvement of the protein kinase Hog1p in the induction of this gene under severe stress conditions, such as high salt. We also detected changes in the...

Induction of the oxidative stress response has been described under many physiological conditions in Saccharomyces cerevisiae, including industrial fermentation for wine yeast biomass production where cells are grown through several batch and fed-batch cultures on molasses. Here, we investigate the influence of aeration on the expression changes of...

Flor yeasts are a particular kind of Saccharomyces cerevisiae strains involved in Sherry wine biological ageing. During this process, yeasts form a film on the wine surface and use ethanol as a carbon source, producing acetaldehyde as a by-product. Acetaldehyde induces BTN2 transcription in laboratory strains. Btn2p is involved in the control of th...

Sulfite treatment is the most common way to prevent grape must spoilage in winemaking because the yeast Saccharomyces cerevisiae is particularly resistant to this chemical. In this paper we report that sulfite resistance depends on sulfur and adenine metabolism. The amount of adenine and methionine in a chemically defined growth medium modulates su...

Physiological capabilities and fermentation performance of Saccharomyces cerevisiae strains to be employed during industrial wine fermentations are critical for the quality of the final product. During the process of biomass propagation, yeast cells are dynamically exposed to a mixed and interrelated group of known stresses such as osmotic, oxidati...

Wine production is a classic example of biotechnology in which 2 organisms, yeast and grape, contribute to the production of an alcoholic beverage. Whilst genetic engineering techniques have been applied to both of these organisms in recent years, much greater progress has been made in modifying yeast. By virtue of the development of GRAS (generall...

We used metabolic engineering to produce wine yeasts with enhanced resistance to glucose deprivation conditions. Glycogen metabolism was genetically modified to overproduce glycogen by increasing the glycogen synthase activity and eliminating glycogen phosphorylase activity. All of the modified strains had a higher glycogen content at the stationar...

The role of trehalose as cell protector against oxidative stress induced by H(2)O(2) has been studied in Saccharomyces cerevisiae mutants in which the two trehalase genes ATH1 and NTH1 are deleted. The addition of low H(2)O(2) concentrations to proliferating cultures of either strain did not harm cell viability and induced a marked activity to Nth1...

When yeast cells are inoculated into grape must for vinification they find stress conditions because of osmolarity, which is due to very high sugar concentration, and pH lower than 4. In this work an analysis of the expression of three osmotic stress induced genes (GPD1, HSP12 and HSP104) under microvinification conditions is shown as a way to prob...

Genetic manipulation of industrial wine yeast strains has become an essential tool for both the study of the molecular mechanisms underlaying their physiology and the improvement of their fermentative properties. The construction of null mutants for any gene in these usually diploid strains, by using a procedure based on sporulation of a heterozygo...

The exceptionally close packing of many yeast genes and other chromosomal elements raises the question of how those elements are functionally insulated. All published work shows that natural insulators are very effective, but transcriptional interference (TI) occurs if they are mutated or if their natural context is altered. Mechanisms to avoid TI...