Influence of flor yeast starters on volatile and nitrogen compounds during a controlled biological aging.

Fino Sherry wine undergoes biological aging carried out by a velum of flor yeast within a traditional dynamic system known as "criaderas and solera". The complex microbiota of biofilm-forming Saccharomyces cerevisiae strains play a crucial role in shaping the distinctive organoleptic profile of these types of wines. For this reason, the aim of this study is to analyze the changes produced by different flor yeast strains in the volatilome and the aminogram of different wines from the criaderas and solera system during biological aging in the laboratory, simulating a flor yeast velum condition at different stages of the system.

Fermentative processes for the upcycling of xylose to xylitol by immobilized cells of WC1507.

Xylitol is a pentose-polyol widely applied in the food and pharmaceutical industry. It can be produced from lignocellulosic biomass, valorizing second-generation feedstocks. Biotechnological production of xylitol requires scalable solutions suitable for industrial scale processes.

Microbial diversity in sherry wine biofilms and surrounding mites.

Sherry wines are film wines produced in the Jerez-Xérès-Sherry and Montilla-Moriles regions in southern Spain which require an aging process under flor biofilms, known as "biological aging". The presence of mites in Sherry wine wineries has been reported and associated with improved wine volatile properties. This work analyzes the microbial diversity in flor biofilms and mites in Sherry wine wineries using Matrix-Assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) and ITS/gene amplification.

Flor yeast immobilization in microbial biocapsules for Sherry wine production: microvinification approach.

Sherry wine is a pale-yellowish dry wine produced in Southern-Spain which features are mainly due to biological aging when the metabolism of biofilm-forming yeasts (flor yeasts) consumes ethanol (and other non-fermentable carbon sources) from a previous alcoholic fermentation, and produces volatile compounds such as acetaldehyde. To start aging and maintain the wine stability, a high alcohol content is required, which is achieved by the previous fermentation or by adding ethanol (fortification). Here, an alternative method is proposed which aims to produce a more economic, distinctive Sherry wine without fortification.

Endogenous CO Overpressure Effect on Higher Alcohols Metabolism during Sparkling Wine Production.

Higher alcohols produced by yeast during the fermentation of sparkling wine must have the greatest impact on the smell and taste of wine. At present, the metabolic response to methanol and higher alcohols formation of under endogenous CO overpressure has not been fully elucidated. In this work, a proteomics and metabolomics approach using a OFFGEL fractionator and the LTQ Orbitrap for the protein identification, followed by a metabolomic study for the detection and quantification of both higher alcohols (GC-FID and SBSE-TD-GC-MS) and amino acids (HPLC), was carried out to investigate the proteomic and metabolomic changes of in relation to higher alcohols formation under a CO overpressure condition in a closed bottle.

Yeast cell vacuum infusion into fungal pellets as a novel cell encapsulation methodology.

Immobilized yeast cells are used industrially in winemaking processes such as sparkling wine and Sherry wine production. Here, a novel approach has been explored for the infusion and immobilization of yeast cells into filamentous fungal pellets, which serve as a porous natural material. This was accomplished through vacuum application to force the yeast cells towards the core of the fungal pellets followed by culture in YPD medium to promote their growth from the interior.

Effect of Bentonite Addition to Pedro Ximénez White Grape Musts before Their Fermentation with Selected Yeasts on the Major Volatile Compounds and Polyols of Wines and Tentative Relationships with the Sensorial Evaluation.

In this work, we study the effect of bentonite addition to the grape must before alcoholic fermentation on the chemical composition and sensorial profile of the obtained wines. Fermentations were carried out with two commercial active dry yeasts treated or not with bentonite and were compared with a control wine obtained by spontaneous fermentation (using the grape must microbiota). Several significant effects on the chemical and sensorial attributes were established by statistical treatments.

Revealing the Yeast Diversity of the Flor Biofilm Microbiota in Sherry Wines Through Internal Transcribed Spacer-Metabarcoding and Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry.

Flor yeast velum is a biofilm formed by certain yeast strains that distinguishes biologically aged wines such as Sherry wine from southern Spain from others. Although is the most common species, 5.8 S-internal transcribed spacer (ITS) restriction fragment length polymorphism analyses have revealed the existence of non- species.

Impact of CO overpressure on yeast mitochondrial associated proteome during the "prise de mousse" of sparkling wine production.

The "prise de mousse" stage during sparkling wine elaboration by the traditional method (Champenoise) involves a second fermentation in a sealed bottle followed by a prolonged aging period, known to contribute significantly to the unique organoleptic properties of these wines. During this stage, CO overpressure, nutrient starvation and high ethanol concentrations are stress factors that affect yeast cells viability and metabolism. Since mitochondria are responsible for energy generation and are required for cell aging and response to numerous stresses, we hypothesized that these organelles may play an essential role during the prise de mousse.

Towards a better understanding of the evolution of odour-active compounds and the aroma perception of sparkling wines during ageing.

A native veil-forming yeast and a commercial yeast strain were used to elaborate sparkling wines by the Champenoise method with a grape variety traditionally used for the production of still wines. Wines aged on lees for fifteen months were sampled at five points and their physicochemical and sensory indices were analysed. Unsupervised and supervised statistical techniques were used to establish a comparison between 81 volatile compounds and eight odour descriptors (chemical, fruity, floral, fatty, balsamic, vegetal, empyreumatic and spicy).

Biological Processes Highlighted in during the Sparkling Wines Elaboration.

Sparkling wines elaboration has been studied by several research groups, but this is the first report on analysis of biological processes according to the Gene Ontology terms (GO terms) and related to proteins expressed by yeast cells during the second fermentation of sparkling wines. This work provides a comprehensive study of the most relevant biological processes in P29, a sparkling wine strain, during the second fermentation under two conditions (without and with endogenous CO overpressure) in the middle and the end of second fermentation. Consequently, a proteomic analysis with the OFFGEL fractionator and protein identification with LTQ Orbitrap XL coupled to HPLC were performed.

Comparative Study of the Proteins Involved in the Fermentation-Derived Compounds in Two Strains of during Sparkling Wine Second Fermentation.

Sparkling wine is a distinctive wine. flor yeasts is innovative and ideal for the sparkling wine industry due to the yeasts' resistance to high ethanol concentrations, surface adhesion properties that ease wine clarification, and the ability to provide a characteristic volatilome and odorant profile. The objective of this work is to study the proteins in a flor yeast and a conventional yeast that are responsible for the production of the volatile compounds released during sparkling wine elaboration.

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

In this study, a first proteomic approach was carried out to characterize the adaptive response of cell wall-related proteins to endogenous CO overpressure, which is typical of second fermentation conditions, in two wine 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.

Using an electronic nose and volatilome analysis to differentiate sparkling wines obtained under different conditions of temperature, ageing time and yeast formats.

Effect of yeast inoculation format (F), temperature (T), and "on lees" ageing time (t) factors were evaluated on the composition of sparkling wines by a quantitative fingerprint obtained from volatile metabolites and the response of an electronic nose (E-nose). Wines elaborated according the traditional method at 10 and 14 °C, free cells and yeast biocapsules formats were monitored at 15 and 24 months of ageing time. Sixty-six volatiles identified and quantified in the eight sampling lots were subjected to a pattern recognition technique.

Autophagic Proteome in Two Strains During Second Fermentation for Sparkling Wine Elaboration.

A correlation between autophagy and autolysis has been proposed in order to acceleratethe acquisition of wine organoleptic properties during sparkling wine elaboration. In this context, aproteomic 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 ofstudying the autophagy-related proteome and comparing the effect of CO2 overpressure duringsparkling wine elaboration. In general, a detrimental effect of pressure and second fermentationdevelopment on autophagy-related proteome was observed in both strains, although it was morepronounced in flor yeast strain G1.

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

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 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.

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 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.

Use of a flor yeast strain for the second fermentation of sparkling wines: Effect of endogenous CO over-pressure on the volatilome.

Saccharomyces cerevisiae flor yeast is used for the first time in sparkling wine-making. Twenty-six oenological variables and fifty-three volatile metabolites are quantified in the middle (P = 3 bar) and at the end (P = 6 bar) of the second fermentation, carried out in open and closed bottles. A heat-map of volatiles and the fingerprints obtained for ten chemical families and ten odorant series visualize the changes for each condition.

Changes in sparkling wine aroma during the second fermentation under CO pressure in sealed bottle.

High quality sparkling wine made by the traditional method requires a second alcoholic fermentation of a base wine in sealed bottles, followed by an aging time in contact with yeast lees. The CO overpressure released during this second fermentation has an important effect on the yeast metabolism and therefore on the wine aroma composition. This study focuses on the changes in chemical composition and 43 aroma compounds released by yeast during this fermentation carried out under two pressure conditions.

Differential Proteome Analysis of a Flor Yeast Strain under Biofilm Formation.

Several strains (flor yeasts) form a biofilm (flor velum) on the surface of Sherry wines after fermentation, when glucose is depleted. This flor velum is fundamental to biological aging of these particular wines. In this study, we identify abundant proteins in the formation of the biofilm of an industrial flor yeast strain.

Use of a flor velum yeast for modulating colour, ethanol and major aroma compound contents in red wine.

The most important and negative effect of the global warming for winemakers in warm and sunny regions is the observed lag between industrial and phenolic grape ripeness, so only it is possible to obtain an acceptable colour when the ethanol content of wine is high. By contrast, the actual market trends are to low ethanol content wines. Flor yeast growing a short time under velum conditions, decreases the ethanol and volatile acidity contents, has a favorable effect on the colour and astringency and significantly changes the wine content in 1-propanol, isobutanol, acetaldehyde, 1,1-diethoxiethane and ethyl lactate.

Functional analysis of stress protein data in a flor yeast subjected to a biofilm forming condition.

In this data article, an OFFGEL fractionator coupled to LTQ Orbitrap XL MS equipment and a SGD filtering were used to detect in a biofilm-forming flor yeast strain, the maximum possible number of stress proteins under the first stage of a biofilm formation conditions (BFC) and under an initial stage of fermentation used as reference, so-called non-biofilm formation condition (NBFC). Protein functional analysis - based on cellular components and biological process GO terms - was performed for these proteins through the SGD Gene Ontology Slim Mapper tool. A detailed analysis and interpretation of the data can be found in "Stress responsive proteins of a flor yeast strain during the early stages of biofilm formation" [1].

Proteins involved in wine aroma compounds metabolism by a Saccharomyces cerevisiae flor-velum yeast strain grown in two conditions.

A proteomic and exometabolomic study was conducted on Saccharomyces cerevisiae flor yeast strain growing under biofilm formation condition (BFC) with ethanol and glycerol as carbon sources and results were compared with those obtained under no biofilm formation condition (NBFC) containing glucose as carbon source. By using modern techniques, OFFGEL fractionator and LTQ-Orbitrap for proteome and SBSE-TD-GC-MS for metabolite analysis, we quantified 84 proteins including 33 directly involved in the metabolism of glycerol, ethanol and 17 aroma compounds. Contents in acetaldehyde, acetic acid, decanoic acid, 1,1-diethoxyethane, benzaldehyde and 2-phenethyl acetate, changed above their odor thresholds under BFC, and those of decanoic acid, ethyl octanoate, ethyl decanoate and isoamyl acetate under NBFC.

Proteins involved in flor yeast carbon metabolism under biofilm formation conditions.

A lack of sugars during the production of biologically aged wines after fermentation of grape must causes flor yeasts to metabolize other carbon molecules formed during fermentation (ethanol and glycerol, mainly). In this work, a proteome analysis involving OFFGEL fractionation prior to LC/MS detection was used to elucidate the carbon metabolism of a flor yeast strain under biofilm formation conditions (BFC). The results were compared with those obtained under non-biofilm formation conditions (NBFC).

A proteomic and metabolomic approach for understanding the role of the flor yeast mitochondria in the velum formation.

Saccharomyces cerevisiae "flor" yeast shows a strong tolerance to high ethanol concentrations and develops a velum (biofilm) on the wine surface after the alcoholic fermentation of grape must. This velum remains along several years during the so called "biological aging" process in the elaboration of some special wines carried out in specific regions around the world and it contributes to the typical organoleptic characteristics of these wines. In order to grow in this condition, flor yeast has to elaborate a response where the mitochondrial function is essential.