Wild Saccharomyces Cerevisiae Strains Research Articles

The Effect of Yeast, Sugar and Sulfur Dioxide on the Volatile Compounds in Wine

This study compares three yeast strains: two wild Saccharomyces cerevisiae strains (Sc1 and Sc5) and a commercial strain (Lc). The objective is to assess their fermentation efficiency and volatile compound production. The factors examined are yeast strain, initial sugar concentration of the must, and the presence of sulfur dioxide. Volatile aroma compounds, determined via GC–MS, were categorized into aromatic series based on aroma descriptors. Out of the volatile compounds analyzed, the yeast strain influenced 39, while sugar content and sulfur dioxide affected 16 and 23 compounds, respectively. Twelve compounds displayed odor activity values exceeding unity, with notable contributions from ethyl esters, β-damascenone, and β-ionone, impacting fruit, floral, and herbal aromatic series. Overall, the Sc1 yeast strain exhibited higher values in the aromatic series compared to the Lc strain. Multivariate analysis revealed that the Sc1 strain highlighted green fruit, citrus, and spice series, while the Lc strain stood out for smoky and herbal aromas. Cluster and principal component analyses emphasized that the aromatic composition of wines produced with wild yeast strains is more influenced by sulfur dioxide than initial sugar content, whereas the opposite holds true for the commercial strain. The key aroma series distinguishing between yeast strains were fruity, green fruit, and citrus for Sc1, and herbal, floral, and smoky for Lc. In conclusion, the Sc1 wild yeast strain showed similar fermentation behavior to the commercial strain, resulting in increased aroma compound presence. The distinctive aromatic profiles contributed by each strain enable winemakers to leverage this diversity and create wines that emphasize specific aromas.

Purification of Yeast Spores to Investigate Their Dynamics of Activation.

Germination is an important developmental process that supports resumption of growth in dormant spores. The study of the mechanisms underlying germination requires a pure spore population devoid of other cell types. This article describes the sporulation of wild Saccharomyces cerevisiae and Saccharomyces paradoxus strains, and the isolation and purification of ascospores. We also describe a method to synchronously induce germination in a spore population as well as to measure spore activation. This procedure can be applied, for example, to the study of environmental conditions that trigger germination. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Sporulation Basic Protocol 2: Spore purification Basic Protocol 3: Germination induction Support Protocol 1: Flow cytometry analysis Support Protocol 2: Heat-shock resistance measurement.

Microencapsulation of Saccharomyces cerevisiae into Alginate Beads: A Focus on Functional Properties of Released Cells.

Five yeast strains (four wild Saccharomyces cerevisiae strains and a collection strain-S. cerevisiae var. boulardii) were encapsulated in alginate beads. Encapsulation yield was at least 60% (100% for some strains) and yeasts survived in beads for 30 days at 4 °C, although the viability was strongly affected during storage at 25 °C (3 log reduction after 7 days). The kinetic of cell release was studied under static and dynamic conditions, but the results suggest that, after 48 h, beads contained a high number of yeasts. Thus, their use is advisable as re-usable carriers of starter cultures or as a vehicle of probiotics into the gut. Finally, some functional properties (biofilm formation, hydrophobicity, auto-aggregation, survival during the transit into the gut) were evaluated on yeasts released by beads to assess if microencapsulation could negatively affect these traits. The results showed that yeasts’ entrapment in beads did not affect probiotic properties.

Ssd1 and the cell wall integrity pathway promote entry, maintenance, and recovery from quiescence in budding yeast.

Wild Saccharomyces cerevisiae strains are typically diploid. When faced with glucose and nitrogen limitation they can undergo meiosis and sporulate. Diploids can also enter a protective, nondividing cellular state or quiescence. The ability to enter quiescence is highly reproducible but shows broad natural variation. Some wild diploids can only enter cellular quiescence, which indicates that there are conditions in which sporulation is lost or selected against. Others only sporulate, but if sporulation is disabled by heterozygosity at the IME1 locus, those diploids can enter quiescence. W303 haploids can enter quiescence, but their diploid counterparts cannot. This is the result of diploidy, not mating type regulation. Introduction of SSD1 to W303 diploids switches fate, in that it rescues cellular quiescence and disrupts the ability to sporulate. Ssd1 and another RNA-binding protein, Mpt5 (Puf5), have parallel roles in quiescence in haploids. The ability of these mutants to enter quiescence, and their long-term survival in the quiescent state, can be rescued by exogenously added trehalose. The cell wall integrity pathway also promotes entry, maintenance, and recovery from quiescence through the Rlm1 transcription factor.

Linkage mapping of yeast cross protection connects gene expression variation to a higher-order organismal trait.

Gene expression variation is extensive in nature, and is hypothesized to play a major role in shaping phenotypic diversity. However, connecting differences in gene expression across individuals to higher-order organismal traits is not trivial. In many cases, gene expression variation may be evolutionarily neutral, and in other cases expression variation may only affect phenotype under specific conditions. To understand connections between gene expression variation and stress defense phenotypes, we have been leveraging extensive natural variation in the gene expression response to acute ethanol in laboratory and wild Saccharomyces cerevisiae strains. Previous work found that the genetic architecture underlying these expression differences included dozens of “hotspot” loci that affected many transcripts in trans. In the present study, we provide new evidence that one of these expression QTL hotspot loci affects natural variation in one particular stress defense phenotype—ethanol-induced cross protection against severe doses of H2O2. A major causative polymorphism is in the heme-activated transcription factor Hap1p, which we show directly impacts cross protection, but not the basal H2O2 resistance of unstressed cells. This provides further support that distinct cellular mechanisms underlie basal and acquired stress resistance. We also show that Hap1p-dependent cross protection relies on novel regulation of cytosolic catalase T (Ctt1p) during ethanol stress in a wild oak strain. Because ethanol accumulation precedes aerobic respiration and accompanying reactive oxygen species formation, wild strains with the ability to anticipate impending oxidative stress would likely be at an advantage. This study highlights how strategically chosen traits that better correlate with gene expression changes can improve our power to identify novel connections between gene expression variation and higher-order organismal phenotypes.

Wild Saccharomyces cerevisiae strains display biofilm-like morphology in contact with polyphenols from grapes and wine

Polyphenols are a major component of wine grapes, and contribute to color and flavor, but their influence upon yeast growth forms has not been investigated. In this work we have studied the effect of polyphenols on the ability of natural isolates of wine-related Saccharomyces cerevisiae strains to form biofilms attaching to plastic surfaces, to grow as mat colonies, to invade media, and to display filamentous growth. The use of carbon- and nitrogen-rich or deficient media simulated grape juice fermentation conditions. The addition of wine polyphenols to these media affected biofilm formation, and cells exhibited a wide variety of invasiveness and mat formation ability with associated different growth and footprint patterns. Microscopic observation revealed that some strains switched to filamentous phenotypes which were able to invade media. The wide range of phenotypic expression observed could have a role in selection of strains suitable for inoculated wine fermentations and may explain the persistence of yeast strains in vineyard and winery environments.

Stress tolerance and growth physiology of yeast strains from the Brazilian fuel ethanol industry

Improved biofuels production requires a better understanding of industrial microorganisms. Some wild Saccharomyces cerevisiae strains, isolated from the fuel ethanol industry in Brazil, present exceptional fermentation performance, persistence and prevalence in the harsh industrial environment. Nevertheless, their physiology has not yet been systematically investigated. Here we present a first systematic evaluation of the widely used industrial strains PE-2, CAT-1, BG-1 and JP1, in terms of their tolerance towards process-related stressors. We also analyzed their growth physiology under heat stress. These strains were evaluated in parallel to laboratory and baker's strains. Whereas the industrial strains performed in general better than the laboratory strains under ethanol or acetic acid stresses and on industrial media, high sugar stress was tolerated equally by all strains. Heat and low pH stresses clearly distinguished fuel ethanol strains from the others, indicating that these conditions might be the ones that mostly exert selective pressure on cells in the industrial environment. During shake-flask cultivations using a synthetic medium at 37°C, industrial strains presented higher ethanol yields on glucose than the laboratory strains, indicating that they could have been selected for this trait-a response to energy-demanding fermentation conditions. These results might be useful to guide future improvements of large-scale fuel ethanol production via engineering of stress tolerance traits in other strains, and eventually also for promoting the use of these fuel ethanol strains in different industrial bioprocesses.

Assessment of competition in wine fermentation among wild Saccharomyces cerevisiae strains isolated from Sangiovese grapes in Tuscany region

In this study we analyzed Saccharomyces cerevisiae isolates obtained from spontaneous fermentations of grapes collected in one vineyard (same rows), during four harvest seasons (2006–2010). A total of 160 isolates were characterized by PCR amplification of inter-δ region. Six strains, representative of the different inter-δ profiles obtained, were tested in single and mixed fermentations at laboratory scale. In all the mixed fermentations, the strain possessing profile A (the only biotype found for two consecutive years) was tested in co-cultures with each of strains showing the other profiles. The strain “A” dominated in almost all the mixed fermentations. The experimental wines obtained from single and co-culture fermentations were analyzed for the content of main secondary compounds. The wines produced by mono-cultures were very different from the wines obtained by co-fermentations, except the wine by strain “A”, which grouped together with mixed-culture wines. Furthermore, the comparison between volatile components determined in mixed culture and blended wines, obtained by mixing monoculture wines, can suggest the existence of yeast metabolic interactions during mixed fermentations. The strain exhibiting the profile A resulted a good “competitor” not only among natural yeast population, but it was also capable to dominate mixed fermentations at lab scale.

Phenotypic selection of a wild Saccharomyces cerevisiae strain for simultaneous saccharification and co-fermentation of AFEX™ pretreated corn stover

BackgroundSimultaneous saccharification and co-fermentation (SSCF) process involves enzymatic hydrolysis of pretreated lignocellulosic biomass and fermentation of glucose and xylose in one bioreactor. The optimal temperatures for enzymatic hydrolysis are higher than the standard fermentation temperature of ethanologenic Saccharomyces cerevisiae. Moreover, degradation products resulting from biomass pretreatment impair fermentation of sugars, especially xylose, and can synergize with high temperature stress. One approach to resolve both concerns is to utilize a strain background with innate tolerance to both elevated temperatures and degradation products.ResultsIn this study, we screened a panel of 108 wild and domesticated Saccharomyces cerevisiae strains isolated from a wide range of environmental niches. One wild strain was selected based on its growth tolerance to simultaneous elevated temperature and AFEX™ (Ammonia Fiber Expansion) degradation products. After engineering the strain with two copies of the Scheffersomyces stipitis xylose reductase, xylitol dehydrogenase and xylulokinase genes, we compared the ability of this engineered strain to the benchmark 424A(LNH-ST) strain in ethanol production and xylose fermentation in standard lab medium and AFEX pretreated corn stover (ACS) hydrolysates, as well as in SSCF of ACS at different temperatures. In SSCF of 9% (w/w) glucan loading ACS at 35°C, the engineered strain showed higher cell viabilities and produced a similar amount of ethanol (51.3 g/L) compared to the benchmark 424A(LNH-ST) strain.ConclusionThese results validate our approach in the selection of wild Saccharomyces cerevisiae strains with thermo-tolerance and degradation products tolerance properties for lignocellulosic biofuel production. The wild and domesticated yeast strains phenotyped in this work are publically available for others to use as genetic backgrounds for fermentation of their pretreated biomass at elevated temperatures.

What do we know about the yeast strains from the Brazilian fuel ethanol industry?

The production of fuel ethanol from sugarcane-based raw materials in Brazil is a successful example of a large-scale bioprocess that delivers an advanced biofuel at competitive prices and low environmental impact. Two to three fed-batch fermentations per day, with acid treatment of the yeast cream between consecutive cycles, during 6-8months of uninterrupted production in a nonaseptic environment are some of the features that make the Brazilian process quite peculiar. Along the past decades, some wild Saccharomyces cerevisiae strains were isolated, identified, characterized, and eventually, reintroduced into the process, enabling us to build up knowledge on these organisms. This information, combined with physiological studies in the laboratory and, more recently, genome sequencing data, has allowed us to start clarifying why and how these strains behave differently from the better known laboratory, wine, beer, and baker's strains. All these issues are covered in this minireview, which also presents a brief discussion on future directions in the field and on the perspectives of introducing genetically modified strains in this industrial process.

Polymorphism detection among wild Saccharomyces cerevisiae strains of different wine origin

In this study, wild Saccharomyces cerevisiae strains, isolated from spontaneously fermenting grapes of different varieties and origins, were submitted to genetic analysis using different molecular techniques, such as amplification of genes coding for cell wall proteins and containing minisatellite-like sequences, karyotyping, mtDNA-RFLP, and analysis of the δ region. The lowest discriminative power was obtained by minisatellites analysis, in particular the amplification of AGA1 genes. Karyotyping and mtDNA-RFLP analysis yielded the same differentiation among the strains, whereas the PCR amplification of δ sequences resulted the best method as it was fast and it showed a very high discriminative power. In any case, it has to be underlined that some strains, showing the same delta profiles, exhibited a different mtDNA restriction profile and electrophoretic karyotype, suggesting that more than one molecular marker is required for reliable strain discrimination. Although the techniques used revealed a different resolution power, they all revealed a genetic relationship among strains isolated from spontaneous fermentation of grapes of different origins. In fact, none of the typing methods was able to discriminate some strains isolated from different areas.

PSI+] Prion Transmission Barriers Protect Saccharomyces cerevisiae from Infection: Intraspecies 'Species Barriers'

[PSI+] is a prion of Sup35p, an essential translation termination and mRNA turnover factor. The existence of lethal [PSI+] variants, the absence of [PSI+] in wild strains, the mRNA turnover function of the Sup35p prion domain, and the stress reaction to prion infection suggest that [PSI+] is a disease. Nonetheless, others have proposed that [PSI+] and other yeast prions benefit their hosts. We find that wild Saccharomyces cerevisiae strains are polymorphic for the sequence of the prion domain and particularly in the adjacent M domain. Here we establish that these variations within the species produce barriers to prion transmission. The barriers are partially asymmetric in some cases, and evidence for variant specificity in barriers is presented. We propose that, as the PrP 129M/V polymorphism protects people from Creutzfeldt-Jakob disease, the Sup35p polymorphisms were selected to protect yeast cells from prion infection. In one prion incompatibility group, the barrier is due to N109S in the Sup35 prion domain and several changes in the middle (M) domain, with either the single N109S mutation or the group of M changes (without the N109S) producing a barrier. In another, the barrier is due to a large deletion in the repeat domain. All are outside the region previously believed to determine transmission compatibility. [SWI+], a prion of the chromatin remodeling factor Swi1p, was also proposed to benefit its host. We find that none of 70 wild strains carry this prion, suggesting that it is not beneficial.

Exploiting Natural Variation inSaccharomyces cerevisiaeto Identify Genes for Increased Ethanol Resistance

Ethanol production from lignocellulosic biomass holds promise as an alternative fuel. However, industrial stresses, including ethanol stress, limit microbial fermentation and thus prevent cost competitiveness with fossil fuels. To identify novel engineering targets for increased ethanol tolerance, we took advantage of natural diversity in wild Saccharomyces cerevisiae strains. We previously showed that an S288c-derived lab strain cannot acquire higher ethanol tolerance after a mild ethanol pretreatment, which is distinct from other stresses. Here, we measured acquired ethanol tolerance in a large panel of wild strains and show that most strains can acquire higher tolerance after pretreatment. We exploited this major phenotypic difference to address the mechanism of acquired ethanol tolerance, by comparing the global gene expression response to 5% ethanol in S288c and two wild strains. Hundreds of genes showed variation in ethanol-dependent gene expression across strains. Computational analysis identified several transcription factor modules and known coregulated genes as differentially expressed, implicating genetic variation in the ethanol signaling pathway. We used this information to identify genes required for acquisition of ethanol tolerance in wild strains, including new genes and processes not previously linked to ethanol tolerance, and four genes that increase ethanol tolerance when overexpressed. Our approach shows that comparative genomics across natural isolates can quickly identify genes for industrial engineering while expanding our understanding of natural diversity.

General factors important for the formation of structured biofilm-like yeast colonies

The lifestyle of wild and laboratory yeast strains significantly differs. In contrast to the smooth colonies of laboratory strains, wild Saccharomyces cerevisiae strains form biofilm-like, strikingly structured colonies possessing distinctive traits enabling them to better survive in hostile environments in the wild. Here, comparing three sets of strains forming differently structured colonies (fluffy, semi-fluffy and smooth), each derived from ancestors with distinct genetic backgrounds isolated from natural settings (BR-88, BR-99 and BR-103), we specified the factors essential for the formation of structured colonies, i.e. for the lifestyle most likely to be preferred in the wild. The ability to form an abundant extracellular matrix (ECM) is one of the features typical for structured colonies. ECM influences colony architecture and many other physiological properties, such as the capability to retain water in a 2-fold surplus to wet cell biomass. ECM composition, however, differs among distinct strains, depending on their particular genetic background. We further show that the expression of certain genes ( AQY1, FLO11) is also strictly related to the particular colony morphology, being highest in the most structured colonies. Flo11p adhesin, important for cell–cell and cell-surface adhesion, is essential for the formation of fluffy colonies and thus significantly contributes to the phenotype variability of wild yeast strains. On the other hand, surprisingly, neither the cell shape nor budding pattern nor the ability to form pseudohyphae directly influences the formation of three-dimensional fluffy colony architecture.

Volatile compounds produced in wine by Colombian wildSaccharomyces cerevisiae strains

Some tropicalSaccharomyces cerevisiae strains, isolated fromChampús, a traditional Colombian low alcoholic fermented beverage, were characterised in order to select yeasts for aroma improvement in wine. H2S production, volatile acidity, β-glucosidase activity, higher alcoholesters and terpenes production were evaluated in this study. These tropical strains were characterised by a considerable production of ethyl hexanoate, 2-phenylethanol, 2-phenylethyl acetate, and geraniol, detected by SPME-GC-MS. Odor activity values were calculated to analyse the effects of yeasts strains on wine aroma, resulting in six distinctive wine groups, as evidenced by discriminant analysis. These results suggest thatSaccharomyces strains isolated fromChampús can be an important source for new tropical yeast biotypes with potential winemaking applications, producing a wide range of aroma compounds.

β-Glucosidase activity in a Saccharomyces cerevisiae wine strain

β-Glucosidase activity contributes to aroma formation during the winemaking process. This study investigated whether β-glucosidase activity was expressed by wild Saccharomyces cerevisiae strains and by a laboratory strain. β-Glucosidase activity was assayed on several culture media and under various growth conditions. The highest activities were obtained in Yeast Extract Peptone medium, but activity was also detected using grape juice as the growth medium, although a 25% drop activity was observed when anaerobic conditions were employed. A number of parameters affecting β-glucosidase activity were evaluated. Optimal conditions for activity were pH 4 and a temperature of 40–50 °C. The results showed β-glucosidase activity to be present during the process of winemaking, although different from the optimal conditions.

Genetic diversity and geographical distribution of wild Saccharomyces cerevisiae strains from the wine-producing area of Charentes, France.

Electrophoretic karyotyping, mitochondrial DNA restriction fragment length polymorphism analysis, and PCR amplification of interspersed repeats were used to study the variability, phylogenetic affinities, and biogeographic distribution of wild Saccharomyces cerevisiae enological yeasts. The survey concentrated on 42 individual wine cellars in the Charentes area (Cognac region, France). A limited number (35) of predominant S. cerevisiae strains responsible for the fermentation process have been identified by the above molecular methods of differentiation. One strain (ACI) was found to be distributed over the entire area surveyed. There seemed to be little correlation between geographic location and genetic affinity.