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A special issue of Fermentation (ISSN 2311-5637). This special issue belongs to the section "Microbial Metabolism, Physiology & Genetics".

Deadline for manuscript submissions: closed (31 July 2022) | Viewed by 28263

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Dr. M. Margarida Baleiras-Couto

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Instituto Nacional de Investigação Agrária e Veterinária – INIAV, Quinta da Almoínha, 2565-191 Dois Portos, Portugal
Interests: yeast diversity; DNA markers; population dynamics; wine yeast

Special Issue Information

Dear Colleagues,

Winemaking involves the sequential development of various species of yeast and bacteria. The production of wine was traditionally based on spontaneous fermentations with yeasts playing the major role. The use of active dry yeast has gradually become common practice.

Information on this microbiota, whether added or indigenous, during the fermentation process is crucial to understand and define wine sensory characteristics.

The study of microbial diversity in the vineyard (soil and plant) and in the winery was first carried out with traditional methods of microbiology, but the introduction of molecular biology techniques, and more recently metagenomics, has broadened the scope and led to the possibility of furthering the study of the microbial structure in wine fermentation. In-depth knowledge of the microbiology of wine and its relationship to the wine metabolome is key to producing unique wines.

Current identification techniques allow the study of microbial diversity and population dynamics at all stages of the winemaking process, including possible microbial contamination during bottling, which is also relevant for the control of the final quality of the wine.

Dr. M. Margarida Baleiras-Couto
Guest Editor

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Keywords

wine
microbiology
yeast
bacteria
population
identification
markers
metagenomic

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Published Papers (10 papers)

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Research

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17 pages, 2873 KiB

Open AccessArticle

A Genome-Wide Phenotypic Analysis of Saccharomyces cerevisiae’s Adaptive Response and Tolerance to Chitosan in Conditions Relevant for Winemaking

by Patrícia Lage, Bárbara B. Coelho, Nuno P. Mira and Ana Mendes-Ferreira

Fermentation 2023, 9(2), 172; https://doi.org/10.3390/fermentation9020172 - 14 Feb 2023

Cited by 1 | Viewed by 2301

Abstract

In the wine industry, the use of chitosan, a non-toxic biodegradable polysaccharide with antimicrobial properties, has been gaining interest with respect to envisaging the reduction in the use of sulfur dioxide (SO₂). Although the mechanisms of toxicity of chitosan against fungal cells have been addressed before, most of the studies undertaken used other sources of chitosan and/or used conditions to solubilize the polymer that were not compatible with winemaking. Herein, the effect of a commercial formulation of chitosan approved for use in winemaking over the growth of the spoilage yeast species Dekkera anomala, Saccharomycodes ludwigii, Zygosaccharomyces bailii, and Pichia anomala was assessed. At the legally allowed concentration of 0.1 g/L, chitosan inhibited the growth of all spoilage yeasts, except for the tested Pichia anomala strains. Interestingly, the highly SO₂-tolerant yeasts S. ludwigii and Z. bailii were highly susceptible to chitosan. The growth of commercial Saccharomyces cerevisiae was also impacted by chitosan, in a strain-dependent manner, albeit at higher concentrations. To dissect this differential inhibitory potential and gain further insight into the interaction of chitosan over fungal cells, we explored a chemogenomic analysis to identify all of the S. cerevisiae genes conferring protection against or increasing susceptibility to the commercial formulation of chitosan. Among the genes found to confer protection against chitosan, a high proportion was found to encode proteins required for the assembly and structuring of the cell wall, enzymes involved in the synthesis of plasma membrane lipids, and components of signaling pathways that respond to damages in the plasma membrane (e.g., the Rim101 pathway). The data obtained also suggest that the fungal ribosome and the vacuolar V-ATPase could be directly targeted by chitosan, since the deletion of genes encoding proteins required for the structure and function of these organelles was found to increase tolerance to chitosan. We also demonstrated, for the first time, that the deletion of ITR1, AGP2 and FPS1, encoding plasma membrane transporters, prominently increased the tolerance of S. cerevisiae to chitosan, suggesting that they can serve as carriers for chitosan. Besides providing new insights into the mode of action of chitosan against wine yeasts, this study adds relevant information for its rational use as a substitute/complementary preservative to SO₂. Full article

(This article belongs to the Special Issue Wine Microbiology)

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18 pages, 3808 KiB

Open AccessArticle

Untargeted Metabolomics Discriminates Grapes and Wines from Two Syrah Vineyards Located in the Same Wine Region

by M. Margarida Baleiras-Couto, Rita Guedes, Filomena L. Duarte, Ana Margarida Fortes and Maria-Luísa Serralheiro

Fermentation 2023, 9(2), 145; https://doi.org/10.3390/fermentation9020145 - 1 Feb 2023

Cited by 3 | Viewed by 2481

Abstract

The influence of terroir in determining wine sensory properties is supported by the specific grape microbiome and metabolome, which provide distinct regional wine characteristics. In this work, the metabolic composition of grapes, must and wine of the Syrah grape variety cultivated on two sites in the same region was investigated. Concomitantly, a sensorial analysis of the produced wines was performed. Ultra-high-resolution liquid chromatography coupled with tandem mass spectrometry (UHPLC-Q-ToF-MS/MS) was applied to identify grape and wine metabolites. Untargeted metabolomics was used to identify putative biomarkers for terroir differentiation. More than 40 compounds were identified, including 28 phenolic compounds and 15 organic acids. The intensity evolution of the analyzed chemical compounds showed similar behavior during the fermentation process in both terroirs. However, the metabolic analysis of the grape, must and wine samples enabled the identification of an anthocyanin, chrysanthemin, as a putative biomarker of terroir 1. The overall sensorial quality of the wines was also evaluated, and according to the hitherto reported results, the wines from site 1 scored better than the wines from site 2. The results highlight the potential of metabolomics to assess grape and wine quality, as well as terroir association. Full article

(This article belongs to the Special Issue Wine Microbiology)

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11 pages, 723 KiB

Open AccessArticle

Effects of Different Aging Methods on the Phenolic Compounds and Antioxidant Activity of Red Wine

by Chao Wang, Chenhui Wang, Ke Tang, Zhiming Rao and Jian Chen

Fermentation 2022, 8(11), 592; https://doi.org/10.3390/fermentation8110592 - 31 Oct 2022

Cited by 8 | Viewed by 2447

Abstract

In this study, oak barrels, glazed pottery altars, unglazed pottery altars, and stainless-steel tanks were selected as aging containers for red wine, and phenolic compounds and antioxidant activity were analyzed and compared. The color of red wine in unglazed pottery altars and glazed pottery altars changed to brick red and brownish yellow, respectively; the color of red wine in oak barrels did not change significantly; and color retention was best in stainless-steel tanks. The total content of anthocyanins and nonanthocyanin phenolic compounds was higher in the unglazed pottery altar group (227.68 mg/L and 288.88 mg/L, respectively) than in the oak barrel group (209.46 mg/L and 273.42 mg/L), the stainless-steel tank group (221.92 mg/L and 213.23 mg/L), or the glazed pottery altar group (74.71 mg/L and 204.43 mg/L). After aging, DPPH (1,1-diphenyl-2-picrylhydrazine free radical scavenging ability), I confirm. (ABTS⁺ free radical scavenging ability), and FRAP (a ferric ion-reducing antioxidant power reduction of Ion Ability) were decreased by 8.8%, 0.5%, and 17.1%, respectively, in the unglazed pottery altar group; by 15.2%, 1.7%, and 19.5%, respectively, in the oak barrel group; by 18.0%, 1.8%, and 20.0%, respectively, in the stainless-steel tank group; and by 18.7%, 4.2%, and 34.9%, respectively, in the glazed pottery altar group. In conclusion, antioxidative ability decreased less in the unglazed pottery altar group than in the other three groups, indicating that unglazed pottery altars retain antioxidant components in red wine well. Full article

(This article belongs to the Special Issue Wine Microbiology)

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14 pages, 1998 KiB

Open AccessArticle

Sexually-Driven Combinatorial Diversity in Native Saccharomyces Wine Yeasts

by Juan Quintero-Blanco, Eugenia Delodi, Andrés Garzón and Juan Jimenez

Fermentation 2022, 8(10), 569; https://doi.org/10.3390/fermentation8100569 - 21 Oct 2022

Cited by 3 | Viewed by 1955

Abstract

Natural diversity represents an inexhaustible source of yeasts for the diversification of wines and the improvement of their properties. In this study, we analysed the genetic diversity of autochthonous Saccharomyces cerevisiae wine yeasts in the Aljarafe of Seville, one of the warmest winemaking regions of Spain. Through multiplex-PCR analysis of five microsatellite markers and RT-PCR determination of the killer genotype, we found 94 different patterns among 150 S. cerevisiae yeast strains isolated from spontaneous fermentation of grape must, thereby representing a highly diverse population. Remarkably, 92% of the isolated strains exhibited high sporulation capacity. Tetrad analysis of sporulating strains rendered a microsatellite marker’s combinatory that mimics patterns observed in the native population, suggesting that the high polymorphism of microsatellite markers found in these wild yeasts might result from sexual reproduction in their natural environment. The identification of unconventional M2/L-A-lus totivirus combinations conferring the killer phenotype also supports this suggestion. One idea behind this study is to determine to what extent the vineyards microbiota in areas with warm climates can provide useful natural yeasts to adapt fermentation processes to the needs imposed by global warming. Analysis of traits of oenological interest in regions potentially affected by global climate changes, such as growth tolerance to ethanol and to sugar stress in the analysed strains, indicated that this broad combinatorial diversity of natural S. cerevisiae yeasts provides a wide range of autochthonous strains with desirable profiles for quality winemaking in warm regions. This combinatorial diversity renders strains with diverse oenological performing abilities. Upon microvinification assays and organoleptic attests, a S. cerevisiae strain with interesting oenological properties has been identified. This result can be considered a successful outcome in industry–academia collaboration. Full article

(This article belongs to the Special Issue Wine Microbiology)

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20 pages, 2654 KiB

Open AccessArticle

Characterization of Saccharomyces Strains Isolated from “Kéknyelű” Grape Must and Their Potential for Wine Production

by Annamária Gerőcs, Tibor Nagy, Katalin Nemes-Barnás, János Májer, Barna Árpád Szőke, Róbert Kővágó, Frederico Magalhães, Brian Gibson, András Szekeres, Ákos Juhász, Katalin Posta and Ferenc Olasz

Fermentation 2022, 8(8), 416; https://doi.org/10.3390/fermentation8080416 - 22 Aug 2022

Cited by 3 | Viewed by 2873

Abstract

Novel wine yeast strains have the potential to satisfy customer demand for new sensorial experiences and to ensure that wine producers have strains that can produce wine as efficiently as possible. In this respect, hybrid yeast strains have recently been the subject of intense research, as they are able to combine the favourable characteristics of both parental strains. In this study, two Saccharomyces “Kéknyelű” grape juice isolates were identified by species-specific PCR and PCR-RFLP methods and investigated with respect to their wine fermentation potential. Physiological characterization of the isolated strains was performed and included assessment of ethanol, sulphur dioxide, temperature and glucose (osmotic stress) tolerance, killer-toxin production, glucose fermentation ability at 16 °C and 24 °C, and laboratory-scale fermentation using sterile “Kéknyelű” must. Volatile components of the final product were studied by gas chromatography (GC) and mass spectrometry (MS). One isolate was identified as a S. cerevisiae × S. kudriavzevii hybrid and the other was S. cerevisiae. Both strains were characterized by high ethanol, sulphur dioxide and glucose tolerance, and the S. cerevisiae strain exhibited the killer phenotype. The hybrid isolate showed good glucose fermentation ability and achieved the lowest residual sugar content in wine. The ester production of the hybrid strain was high compared to the control S. cerevisiae starter strain, and this contributed to the fruity aroma of the wine. Both strains have good oenological characteristics, but only the hybrid yeast has the potential for use in wine fermentation. Full article

(This article belongs to the Special Issue Wine Microbiology)

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10 pages, 1105 KiB

Open AccessArticle

Comparative Evaluation of Secreted Plant Carotenoid Cleavage Dioxygenase 1 (CCD1) Enzymes in Saccharomyces cerevisiae

by John J. B. Timmins, Heinrich Kroukamp, Roy S. K. Walker, Isak S. Pretorius and Ian T. Paulsen

Fermentation 2022, 8(8), 395; https://doi.org/10.3390/fermentation8080395 - 15 Aug 2022

Cited by 3 | Viewed by 2259

Abstract

Enabling technologies in synthetic biology now present the opportunity to engineer wine yeast for enhanced novel aromas. In doing so, improved wine products will increase the desirability of wine for the consumer and add value to the winemaker. The action of the enzyme carotenoid cleavage dioxygenase 1 (CCD1) on β-carotene to produce β-ionone is of interest to improve the aroma and flavour of the wine. Engineering the yeast, Saccharomyces cerevisiae, to produce higher concentrations of CCD1 in grape-must presents an opportunity to increase the levels of this volatile organic compound, thus enhancing the organoleptic properties of wine. To this end, four phylogenetically diverse plant CCD1 genes were synthesised with a secretion signal peptide and transformed into S. cerevisiae. The relative ability of each enzyme secreted into the yeast supernatant to cleave the deep orange C₄₀ β-carotene was determined by spectrophotometry; furthermore, the by-product of such cleavage, the highly aromatic C₁₃ β-ionone, was assessed by head-space solid-phase micro-extraction, with analysis and detection by GCMS. Reduction in β-carotene levels and release of β-ionone from the supernatant were validated by LCMS detection of CCD1. These experiments demonstrated that expression in yeast of the CCD1s derived from Petunia hybrida and Vitis vinifera and their subsequent secretion into the medium provided superior efficacy in both β-carotene reduction and β-ionone liberation. We anticipate this knowledge being of benefit to future winemakers in producing a vinous product with enhanced organoleptic properties. Full article

(This article belongs to the Special Issue Wine Microbiology)

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13 pages, 2777 KiB

Open AccessArticle

Integrated Fertilization with Bagasse Vermicompost Changes the Microbiome of Mencía Must and Wine

by Daniela Rosado, Marta Lores, Ignacio Ramos-Tapia, Keith A. Crandall, Marcos Pérez-Losada and Jorge Domínguez

Fermentation 2022, 8(8), 357; https://doi.org/10.3390/fermentation8080357 - 27 Jul 2022

Cited by 6 | Viewed by 2209

Abstract

Winemaking is a well-known process that includes several steps in the production of grape must and wine. Grape marc, or bagasse, is a byproduct of wine production that can be vermicomposted and used as organic fertilizer. Grape marc vermicompost has microbial communities that are richer and more stable than grape marc alone, and its addition to a vineyard’s terroir can improve grape yields and wine quality. Here we compare the must and wine microbiota of Mencía from grapevines treated with and without (standard fertilization) vermicompost derived from Mencía grape marc. Mencía is a high-quality red wine broadly grown in Galicia, Spain, and is appreciated for its fresh acidity and fruity flavors. When Mencía grapevines are treated with vermicompost derived from its grape marc, Mencía vines increase their grape production, and the final wine improves its organoleptic properties. Metataxonomic analyses of the bacterial 16S rRNA and fungal ITS gene regions showed that Mencía must and wine have the distinct taxonomic composition (phyla, genera and ASVs—amplicon sequence variants) of bacterial and fungal groups. Must and wine bacteriotas and mycobiotas show no significant variation in alpha-diversity, while wine bacteriotas and mycobiotas show significant differences in microbial structure (beta-diversity) between treated and control grapevines. Likewise, the functional diversity and predicted metabolic pathways (biosynthesis, degradation/utilization/assimilation, generation of precursor metabolites and energy, macromolecule modification and superpathways) of the must and wine microbiota also show significant changes. Our study proposes that changes in the abundance of microbial taxa and the metabolic processes they undergo during winemaking may improve Mencía’s organoleptic properties and productivity. Full article

(This article belongs to the Special Issue Wine Microbiology)

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14 pages, 1040 KiB

Open AccessArticle

Ecological Distribution and Oenological Characterization of Native Saccharomyces cerevisiae in an Organic Winery

by Alice Agarbati, Laura Canonico, Francesca Comitini and Maurizio Ciani

Fermentation 2022, 8(5), 224; https://doi.org/10.3390/fermentation8050224 - 13 May 2022

Cited by 7 | Viewed by 2572

Abstract

The relation between regional yeast biota and the organoleptic characteristics of wines has attracted growing attention among winemakers. In this work, the dynamics of a native Saccharomyces cerevisiae population was investigated in an organic winery. In this regard, the occurrence and the persistence of native S. cerevisiae were evaluated in the vineyard and winery and during spontaneous fermentation of two nonconsecutive vintages. From a total of 98 strains, nine different S. cerevisiae biotypes were identified that were distributed through the whole winemaking process, and five of them persisted in both vintages. The results of the oenological characterization of the dominant biotypes (I and II) show a fermentation behavior comparable to that exhibited by three common commercial starter strains, exhibiting specific aromatic profiles. Biotype I was characterized by some fruity aroma compounds, such as isoamyl acetate and ethyl octanoate, while biotype II was differentiated by ethyl hexanoate, nerol, and β-damascenone production also in relation to the fermentation temperature. These results indicate that the specificity of these resident strains should be used as starter cultures to obtain wines with distinctive aromatic profiles. Full article

(This article belongs to the Special Issue Wine Microbiology)

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17 pages, 3207 KiB

Open AccessArticle

New Insights into the Origin of Volatile Sulfur Compounds during Wine Fermentation and Their Evolution during Aging

by Rafael Jiménez-Lorenzo, Vincent Farines, Jean-Marie Sablayrolles, Carole Camarasa and Audrey Bloem

Fermentation 2022, 8(4), 139; https://doi.org/10.3390/fermentation8040139 - 23 Mar 2022

Cited by 4 | Viewed by 3337

Abstract

Volatile sulfur compounds (VSCs) are associated with unpleasant reductive aromas and are responsible for an important reduction in wine quality, causing major economic losses. Understanding the origin of these compounds in wine remains a challenge, as their formation and further evolution during winemaking can involve both chemical and biological reactions. Comparing the VSCs profile (i) of fermenting synthetic grape juices supplemented with a selected VSC (eight compounds tested) and incubated in presence or absence of yeast, and (ii) during storage of wines under an accelerated aging procedure, allowed us to elucidate the chemical and metabolic connections between VSCs during fermentation and aging. Yeast metabolism, through the Ehrlich pathway and acetylation reactions, makes an important contribution to the formation of compounds such as methionol, 3-methylthiopropionate, 3-methylthiopropylacetate, 3-mercaptopropanol, 2-mercaptoethanol and thioesters. By contrast, chemical reactions are responsible for interconversions between thiols and disulfides, the formation of thiols from thioesters or, more surprisingly, the formation of ethylthiopropanol from methionol during fermentation. During aging, variations in heavy VSC concentrations, such as an increase in 3-methylthiopropylacetate and a decrease in ethyl-3-methylthiopropionate formation, were evidenced. Overall, this study highlights that it is essential to consider both yeast metabolism and the high chemical reactivity of VSCs to understand their formation and evolution during winemaking. Full article

(This article belongs to the Special Issue Wine Microbiology)

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Review

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21 pages, 4435 KiB

Open AccessReview

Sherry Wines: Worldwide Production, Chemical Composition and Screening Conception for Flor Yeasts

by Daria Avdanina and Alexander Zghun

Fermentation 2022, 8(8), 381; https://doi.org/10.3390/fermentation8080381 - 10 Aug 2022

Cited by 8 | Viewed by 3994 | Correction

Abstract

The manufacturing of sherry wines is a unique, carefully regulated process, from harvesting to quality control of the finished product, involving dynamic biological aging in a “criadera-solera” system or some other techniques. Specialized “flor” strains of the yeast Saccharomyces cerevisiae play the central role in the sherry manufacturing process. As a result, sherry wines have a characteristic and unique chemical composition that determines their organoleptic properties (such as color, odor, and taste) and distinguishes them from all other types of wine. The use of modern methods of genetics and biotechnology contributes to a deep understanding of the microbiology of sherry production and allows us to define a new methodology for breeding valuable flor strains. This review discusses the main sherry-producing regions and the chemical composition of sherry wines, as well as genetic, oenological, and other selective markers for flor strains that can be used for screening novel candidates that are promising for sherry production among environmental isolates. Full article

(This article belongs to the Special Issue Wine Microbiology)

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