First Proteomic Approach to Identify Cell Death Biomarkers in Wine Yeasts during Sparkling Wine Production

Apoptosis and later autolysis are biological processes which take place in Saccharomyces cerevisiae during industrial fermentation processes, which involve costly and time-consuming aging periods. Therefore, the identification of potential cell death biomarkers can contribute to the creation of a long-term strategy in order to improve and accelerate the winemaking process. Here, we performed a proteomic analysis based on the detection of possible apoptosis and autolysis protein biomarkers in two industrial yeast strains commonly used in post-fermentative processes (sparkling wine secondary fermentation and biological aging) under typical sparkling wine elaboration conditions. Pressure had a negatively effect on viability for flor yeast, whereas the sparkling wine strain seems to be more adapted to these conditions. Flor yeast strain experienced an increase in content of apoptosis-related proteins, glucanases and vacuolar proteases at the first month of aging. Significant correlations between viability and apoptosis proteins were established in both yeast strains. Multivariate analysis based on the proteome of each process allowed to distinguish among samples and strains. The proteomic profile obtained in this study could provide useful information on the selection of wine strains and yeast behavior during sparkling wine elaboration. Additionally, the use of flor yeasts for sparkling wine improvement and elaboration is proposed.

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Differential Analysis of Proteins Involved in Ester Metabolism in two Saccharomyces cerevisiae Strains during the Second Fermentation in Sparkling Wine Elaboration

Microorganisms ◽

10.3390/microorganisms8030403 ◽

2020 ◽

Vol 8 (3) ◽

pp. 403 ◽

Cited By ~ 2

Author(s):

Maria del Carmen González-Jiménez ◽

Jaime Moreno-García ◽

Teresa García-Martínez ◽

Juan José Moreno ◽

Anna Puig-Pujol ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Protein Identification ◽

Ethyl Esters ◽

Stir Bar Sorptive Extraction ◽

Biological Aging ◽

Yeast Fermentation ◽

Sparkling Wine ◽

Wine Production ◽

Yeast Strains ◽

Flor Yeast

The aromatic metabolites derived from yeast metabolism determine the characteristics of aroma and taste in wines, so they are considered of great industrial interest. Volatile esters represent the most important group and therefore, their presence is extremely important for the flavor profile of the wine. In this work, we use and compare two Saccharomyces cerevisiae yeast strains: P29, typical of sparkling wines resulting of second fermentation in a closed bottle; G1, a flor yeast responsible for the biological aging of Sherry wines. We aimed to analyze and compare the effect of endogenous CO2 overpressure on esters metabolism with the proteins related in these yeast strains, to understand the yeast fermentation process in sparkling wines. For this purpose, protein identification was carried out using the OFFGEL fractionator and the LTQ Orbitrap, following the detection and quantification of esters with gas chromatograph coupled to flame ionization detector (GC-FID) and stir-bar sorptive extraction, followed by thermal desorption and gas chromatography-mass spectrometry (SBSE-TD-GC-MS). Six acetate esters, fourteen ethyl esters, and five proteins involved in esters metabolism were identified. Moreover, significant correlations were established between esters and proteins. Both strains showed similar behavior. According to these results, the use of this flor yeast may be proposed for the sparkling wine production and enhance the diversity and the typicity of sparkling wine yeasts.

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A Differential Proteomic Approach to Characterize the Cell Wall Adaptive Response to CO2 Overpressure during Sparkling Wine-Making Process

Microorganisms ◽

10.3390/microorganisms8081188 ◽

2020 ◽

Vol 8 (8) ◽

pp. 1188

Author(s):

Juan Antonio Porras-Agüera ◽

Juan Carlos Mauricio ◽

Jaime Moreno-García ◽

Juan Moreno ◽

Teresa García-Martínez

Keyword(s):

Cell Wall ◽

Adaptive Response ◽

Wine Yeast ◽

Sparkling Wine ◽

Cell Wall Proteins ◽

Protein Biomarkers ◽

Wine Making ◽

Proteomic Approach ◽

Flor Yeast ◽

Related Proteins

In this study, a first proteomic approach was carried out to characterize the adaptive response of cell wall-related proteins to endogenous CO2 overpressure, which is typical of second fermentation conditions, in two wine Saccharomyces cerevisiae strains (P29, a conventional second fermentation strain, and G1, a flor yeast strain implicated in sherry wine making). The results showed a high number of cell wall proteins in flor yeast G1 under pressure, highlighting content at the first month of aging. The cell wall proteomic response to pressure in flor yeast G1 was characterized by an increase in both the number and content of cell wall proteins involved in glucan remodeling and mannoproteins. On the other hand, cell wall proteins responsible for glucan assembly, cell adhesion, and lipid metabolism stood out in P29. Over-represented proteins under pressure were involved in cell wall integrity (Ecm33p and Pst1p), protein folding (Ssa1p and Ssa2p), and glucan remodeling (Exg2p and Scw4p). Flocculation-related proteins were not identified under pressure conditions. The use of flor yeasts for sparkling wine elaboration and improvement is proposed. Further research based on the genetic engineering of wine yeast using those genes from protein biomarkers under pressure alongside the second fermentation in bottle is required to achieve improvements.

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Autophagic Proteome in Two Saccharomyces cerevisiae Strains during Second Fermentation for Sparkling Wine Elaboration

Microorganisms ◽

10.3390/microorganisms8040523 ◽

2020 ◽

Vol 8 (4) ◽

pp. 523 ◽

Cited By ~ 1

Author(s):

Juan Antonio Porras-Agüera ◽

Jaime Moreno-García ◽

María del Carmen González-Jiménez ◽

Juan Carlos Mauricio ◽

Juan Moreno ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Multivariate Data Analysis ◽

Detrimental Effect ◽

Sparkling Wine ◽

Design Strategies ◽

Autophagosome Formation ◽

Organoleptic Properties ◽

Time Production ◽

Flor Yeast ◽

Wine Strain

A correlation between autophagy and autolysis has been proposed in order to accelerate the acquisition of wine organoleptic properties during sparkling wine elaboration. In this context, a proteomic analysis was carried out in two industrial Saccharomyces cerevisiae strains (P29, conventional sparkling wine strain and G1, implicated in sherry wine elaboration) with the aim of studying the autophagy-related proteome and comparing the effect of CO2 overpressure during sparkling wine elaboration. In general, a detrimental effect of pressure and second fermentation development on autophagy-related proteome was observed in both strains, although it was more pronounced in flor yeast strain G1. Proteins mainly involved in autophagy regulation and autophagosome formation in flor yeast G1, and those required for vesicle nucleation and expansion in P29 strain, highlighted in sealed bottle. Proteins Sec2 and Sec18 were detected 3-fold under pressure conditions in P29 and G1 strains, respectively. Moreover, ‘fingerprinting’ obtained from multivariate data analysis established differences in autophagy-related proteome between strains and conditions. Further research is needed to achieve more solid conclusions and design strategies to promote autophagy for an accelerated autolysis, thus reducing cost and time production, as well as acquisition of good organoleptic properties.

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Enological Behaviour of Biofilms Formed by Genetically-Characterized Strains of Sherry Yeast

The Open Biotechnology Journal ◽

10.2174/1874070701307010023 ◽

2013 ◽

Vol 7 (1) ◽

pp. 23-29 ◽

Cited By ~ 10

Author(s):

María E. Rodríguez ◽

Juan J. Infante ◽

Juan J. Mesa ◽

Laureana Rebordinos ◽

Jesús M. Cantoral

Keyword(s):

Wine Industry ◽

Biological Aging ◽

Yeast Strains ◽

Sherry Wine ◽

Flor Yeast ◽

Film Forming ◽

Pure Cultures ◽

Biofilm Development ◽

Kinetics Of ◽

Strain Dependent

The flor yeasts (Saccharomyces cerevisiae) form a biofilm, known as flor velum, on the surface of fino-type sherry wine at the end of the alcoholic fermentation. These film-forming yeasts are responsible for the oxidative transformation of alcohol to acetaldehyde, together with other reactions, which produce the characteristic flavours and aromas of these wines. In this study, we examine the enological behaviour of eight flor yeast strains biofilms in biological aging experiments carried out in the laboratory. Strains with identical chromosomal and mitochondrial DNA patterns and the same origin showed a more closely-related enological behaviour. But the kinetics of growth and acetaldehyde accumulation in the wine were found to be strain-dependent. Moreover, some strains were marked by high acetaldehyde accumulation in their pure cultures during the various phases of the biofilm development. These results provide valuable knowledge for planning technical strategies to improve the biological aging process in the sherry wine industry.

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Application of yeast with reduced alcohol yield for sparkling wine production

BIO Web of Conferences ◽

10.1051/bioconf/20191202021 ◽

2019 ◽

Vol 12 ◽

pp. 02021 ◽

Cited By ~ 1

Author(s):

M. Schmitt ◽

S. Broschart ◽

C.-D. Patz ◽

D. Rauhut ◽

M. Friedel ◽

...

Keyword(s):

Yeast Strain ◽

Wine Industry ◽

Sparkling Wine ◽

Alcohol Content ◽

Wine Production ◽

Alcohol Production ◽

Volatile Acidity ◽

Yeast Strains ◽

Lower Alcohol ◽

Commercial Yeast

Two commercial yeast strains with reduced alcohol production in comparison with a commercial yeast strain with common alcohol yield were assed for their suitability in sparkling wine production according to the traditional bottle fermentation. The different yeast strains were applied for the first fermentation. As expected the base wine differed in terms alcohol. Furthermore the yeast with lower alcohol content showed higher values of glycerol, higher arginine content and in the same time reduced levels of proline after fermentation. However those samples showed increased volatile acidity values, compared to the control wines. The later bottle fermentation with a uniform yeast strain showed similar fermentation kinetics for all four lots. Sensory evaluation showed no clear differences between the sparkling wines that were stored 9 months on the lees. The base wines nevertheless clearly differed from each other. Besides the increased production of volatile acidity, the tested yeast strains with lower alcohol production appear very promising for the sparkling wine industry to face the generally rising alcohol contents worldwide.

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Autophagy gene overexpression in Saccharomyces cerevisiae perturbs subcellular organellar function and accumulates ROS to accelerate cell death with relevance to sparkling wine production

Applied Microbiology and Biotechnology ◽

10.1007/s00253-018-9304-y ◽

2018 ◽

Vol 102 (19) ◽

pp. 8447-8464 ◽

Cited By ~ 4

Author(s):

Richard Preiss ◽

Caroline Tyrawa ◽

George van der Merwe

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Death ◽

Gene Overexpression ◽

Sparkling Wine ◽

Wine Production ◽

Autophagy Gene

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Biological Aging of Sherry Wines Using Pure Cultures of Two Flor Yeast Strains under Controlled Microaeration

Journal of Agricultural and Food Chemistry ◽

10.1021/jf0478713 ◽

2005 ◽

Vol 53 (13) ◽

pp. 5258-5264 ◽

Cited By ~ 25

Author(s):

David Muñoz ◽

Rafael A. Peinado ◽

Manuel Medina ◽

Juan Moreno

Keyword(s):

Biological Aging ◽

Yeast Strains ◽

Flor Yeast ◽

Pure Cultures

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Non-Conventional Grape Varieties and Yeast Starters for First and Second Fermentation in Sparkling Wine Production Using the Traditional Method

Fermentation ◽

10.3390/fermentation7040321 ◽

2021 ◽

Vol 7 (4) ◽

pp. 321

Author(s):

María Laura Raymond Eder ◽

Alberto Luis Rosa

Keyword(s):

Traditional Method ◽

Pinot Noir ◽

Sparkling Wine ◽

Key Factors ◽

Wine Production ◽

Factors Affecting ◽

Yeast Strains ◽

Fundamental Research ◽

Sensory Profiles ◽

Grape Varieties

Sparkling wine production using the traditional method involves a second fermentation of still wines in bottles, followed by prolonged aging on lees. The key factors affecting the organoleptic profiles of these wines are the grape varieties, the chemical and sensory attributes of the base wines elaborated, the yeast strains used for first and second fermentation, and the winery practices. While Chardonnay and Pinot noir are gold standard grape varieties in sparkling wine production, other valuable grape cultivars are used worldwide to elaborate highly reputable sparkling wines. Fundamental research on the chemical and sensory profiles of innovative sparkling wines produced by the traditional method, using non-conventional grape varieties and novel yeast strains for first and/or second fermentation, is accompanying their market diversification. In this review, we summarize relevant aspects of sparkling wine production using the traditional method and non-conventional grape varieties and yeast starters.

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Differential Response of the Proteins Involved in Amino Acid Metabolism in Two Saccharomyces cerevisiae Strains during the Second Fermentation in a Sealed Bottle

Applied Sciences ◽

10.3390/app112412165 ◽

2021 ◽

Vol 11 (24) ◽

pp. 12165

Author(s):

María del Carmen González-Jiménez ◽

Juan Carlos Mauricio ◽

Jaime Moreno-García ◽

Anna Puig-Pujol ◽

Juan Moreno ◽

...

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Amino Acid Metabolism ◽

Acid Metabolism ◽

Sparkling Wine ◽

Sulfur Amino Acids ◽

Wine Production ◽

Yeast Strains ◽

The Difference ◽

Lysine Metabolism

The traditional method for sparkling wine making consists of a second fermentation of a base wine followed by ageing in the same bottle that reaches the consumers. Nitrogen metabolism is the second most important process after carbon and takes place during wine fermentation by yeast. Amino acids are the most numerous nitrogen compounds released by this process. They contribute to the organoleptic properties of the wines and, therefore, to their sensory quality. The main objective of this study is to compare the differential proteomic response of amino acid metabolism, specifically their proteins and their interactions in the G1 strain (unconventional yeast) during sparkling wine production versus the conventional P29 strain. One of the new trends in winemaking is the improvement of the organoleptic diversity of wine. We propose the use of unconventional yeast that shows desirable characteristics for the industry. For this purpose, these two yeasts were grown at sealed bottle conditions for the second fermentation (Champenoise method). No differences were obtained in the middle of fermentation between the yeast strains. The number of proteins identified, and the relationships established, were similar, highlighting lysine metabolism. At the end of the second fermentation, the difference between each strain was remarkable. Hardly any proteins were identified in unconventional versus conventional yeast. However, in both strains, the metabolism of sulfur amino acids, methionine, and cysteine obtained a greater number of proteins involved in these processes. The release of these amino acids to the medium would allow the yeast to balance the internal redox potential by reoxidation of NADPH. This study is focused on the search for a more complete knowledge of yeast metabolism, specifically the metabolism of amino acids, which are key compounds during the second fermentation.

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