Quantitative Data-Independent Acquisition Glycoproteomics of Sparkling Wine.

Sparkling wine is an alcoholic beverage enjoyed around the world. The sensory properties of sparkling wine depend on a complex interplay between the chemical and biochemical components in the final product. Glycoproteins have been linked to positive and negative qualities in sparkling wine, but the glycosylation profiles of sparkling wine have not been previously investigated in detail. We analyzed the glycoproteome of sparkling wines using protein- and glycopeptide-centric approaches. We developed an automated workflow that created ion libraries to analyze sequential window acquisition of all theoretical mass spectra data-independent acquisition mass spectrometry data based on glycopeptides identified by Byonic (Protein Metrics; version 2.13.17). We applied our workflow to three pairs of experimental sparkling wines to assess the effects of aging on lees and of different yeast strains used in the liqueur de tirage for secondary fermentation. We found that aging a cuvée on lees for 24 months compared with 8 months led to a dramatic decrease in overall protein abundance and an enrichment in large glycans at specific sites in some proteins. Secondary fermentation of a Riesling wine with Saccharomyces cerevisiae yeast strain Siha4 produced more yeast proteins and glycoproteins than with S. cerevisiae yeast strain DV10. The abundance and glycosylation profiles of grape glycoproteins were also different between grape varieties. To our knowledge, this work represents the first in-depth study into protein- and peptide-specific glycosylation in sparkling wines and describes a quantitative glycoproteomic sequential window acquisition of all theoretical mass spectra/data-independent acquisition workflow that is broadly applicable to other sample types.

Contribution of terpenes, methanethiol, and fermentative esters to sparkling wine aroma in relation to production technology, vintage, and aging: a case study on Durello wines.

BACKGROUND: The present study aimed to assess the chemical and sensory variability existing across sparkling white wines of a single appellation in relation to production technique and aging, identifying the existence of specific aroma chemical and sensory (odor) signatures. For this purpose, we selected Durello, an emerging sparkling white wine from the Veneto region produced using either Charmat or Classico methods. RESULTS: Comparison of Durello varietal volatiles profile with other Italian sparkling and still wines highlighted specific features of Durello, such as high content of 1,4-cineole and non-megastigmane norisoprenoids. The production technique (Charmat or Champenoise) impacted both the volatile chemical profile, influencing the content of esters, fatty acids, cyclic terpenoids, C6 alcohols, ß-damascenone and methanethiol. With regard to wine age and vintage, an influence on esters, p-menthane-1,8-diol, cineoles and methyl salicylate was observed. CONCLUSION: Sensory evaluation by means of a sorting task approach indicated the existence of two main types of odor profile, one clearly associated with Classico method. The main drivers of such aroma diversity were esters, 1,4-cineole and methanethiol. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Differentiation through E-nose and GC-FID data modeling of rosé sparkling wines elaborated via traditional and Charmat methods.

BACKGROUND: The growing demand for rosé sparkling wine has led to an increase in its production. Traditional or Charmat wine-making influence the aromatic profiles in wine. An analysis such as gas chromatography makes an accurate assessment of wines based on volatile detection but is resource intensive. On the other hand, the electronic nose (E-nose) has emerged as a versatile tool, offering rapid, cost-effective discrimination of wines, and contributing insights into quality and production processes because of its aptitude to perform a global aromatic pattern evaluation. In the present study, rosé sparkling wines were produced using both methods and major volatile compounds and polyols were measured. Wines were tested by E-nose and predictive modelling was performed to distinguish them. RESULTS: Volatile profiles showed differences between Charmat and traditional methods, especially at 5 months of aging. A partial least square discriminant analysis (PLS-DA) was carried out on E-nose detections, obtaining a model that describes 94% of the variability, separating samples in different clusters and correctly identifying different classes. The differences derived from PLS-DA clustering agree with the results obtained by gas-chromatography. Moreover, a principal components regression model was built to verify the ability of the E-nose to non-destructively predict the amount of different volatiles analyzed. CONCLUSION: Production methods of Rosé sparkling wine affect the final wine aroma profiles as a result of the differences in terms of volatiles. The PLS-DA of the data obtained with E-nose reveals that distinguishing between Charmat and traditional methods is possible. Moreover, predictive models using gas chromatography-flame ionization detection analysis and E-nose highlight the possibility of fast and efficient prediction of volatiles from the E-nose. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Data Fusion Approaches for the Characterization of Musts and Wines Based on Biogenic Amine and Elemental Composition.

Samples from various winemaking stages of the production of sparkling wines using different grape varieties were characterized based on the profile of biogenic amines (BAs) and the elemental composition. Liquid chromatography with fluorescence detection (HPLC-FLD) combined with precolumn derivatization with dansyl chloride was used to quantify BAs, while inductively coupled plasma (ICP) techniques were applied to determine a wide range of elements. Musts, base wines, and sparkling wines were analyzed accordingly, and the resulting data were subjected to further chemometric studies to try to extract information on oenological practices, product quality, and varieties. Although good descriptive models were obtained when considering each type of data separately, the performance of data fusion approaches was assessed as well. In this regard, low-level and mid-level approaches were evaluated, and from the results, it was concluded that more comprehensive models can be obtained when joining data of different natures.

Influence of vinification process over the composition of volatile compounds and sensorial characteristics of greek wines.

Wine is one of the most traditional consumed alcoholic beverages in the world. Millions of wine enthusiasts worldwide duly appreciate a wine of excellent quality in terms of physicochemical and sensorial properties. Different classes of wines have different volatile compositions and sensorial properties, which can be altered, depending on the vinification process and use of additives. Among the widely employed additives in wine production is sulfite (SO2). The popularity of sulfite lies in its ability to preserve the flavor and freshness of wine; however, depending on the quantity, sulfite can pose serious health risks to consumers and affect the quality of the drink. The present study evaluated and compared the compositions of volatile compounds and sensorial properties of sparkling and traditional wines (with and without SO2) produced from Greek grapes 'Grechetto', 'Greco bianco' and 'Greco di tufo'. The results obtained of the composition of volatile compounds in these wines pointed to differences between SO2-containing, SO2-free and sparkling wines, with different amounts of compounds such as alcohols, esters, fatty acids, phenols and differences in sensorial properties. The 'Grechetto' wine, prepared without SO2, exhibited greater quality, better volatile composition, and better sensorial properties compared to the wines produced with SO2. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13197-021-05160-y.

Development and optimization of a loop-mediated isothermal amplification (LAMP) assay for the species-specific detection of Penicillium expansum.

Penicillium expansum is the main cause of Blue Mold Decay, which is the economically most significant postharvest disease on fruits. It occurs especially on pomaceous fruits such as apples and pears but also on a wide range of other fruits such as grapes or strawberries. Besides its negative economic effects on the industry, the fungus is also of health concern as it produces patulin, a mycotoxin known to provoke harmful effects in humans. A specific and rapid detection of this fungus therefore is required. In the current study, a loop-mediated isothermal amplification (LAMP) assay was developed and optimized for the species-specific detection of P. expansum. The assay showed high specificity during tests with genomic DNA of 187 fungal strains. The detection limit of the developed assay was 25 pg genomic DNA of P. expansum per reaction. The assay was successfully applied for the detection of the fungus on artificially contaminated apples, grapes, apple juice, apple puree, and grape juice. The developed assay is a promising tool for rapid, sensitive, specific, and cost-efficient detection of P. expansum in quality control applications in the food and beverage industry.

Novel Methods to Manipulate Autolysis in Sparkling Wine: Effects on Yeast.

Sparkling wine made by the traditional method (Méthode Traditionelle) develops a distinct and desirable flavour and aroma profile attributed to proteolytic processes during prolonged ageing on lees. Microwave, ultrasound and addition of ß-glucanase enzymes were applied to accelerate the disruption of Saccharomyces cerevisiae, and added to the tirage solution for secondary fermentation in traditional sparkling winemaking. Scanning electron microscopy and flow cytometry analyses were used to observe and describe yeast whole-cell anatomy, and cell integrity and structure via propidium iodide (PI) permeability after 6-, 12- and 18-months post-tirage. Treatments applied produced features on lees that were distinct from that of the untreated control yeast. Whilst control yeast displayed budding cells (growth features) with smooth, cavitated and flat external cell appearances; microwave treated yeast cells exhibited modifications like 'doughnut' shapes immediately after treatment (time 0). Similar 'doughnut'-shaped and 'pitted/porous' cell features were observed on progressively older lees from the control. Flow cytometry was used to discriminate yeast populations; features consistent with cell disruption were observed in the microwave, ultrasound and enzyme treatments, as evidenced by up to 4-fold increase in PI signal in the microwave treatment. Forward and side scatter signals reflected changes in size and structure of yeast cells, in all treatments applied. When flow cytometry was interpreted alongside the scanning electron microscopy images, bimodal populations of yeast cells with low and high PI intensities were revealed and distinctive 'doughnut'-shaped cell features observed in association with the microwave treatment only at tirage, that were not observed until 12 months wine ageing in older lees from the control. This work offers both a rapid approach to visualise alterations to yeast cell surfaces and a better understanding of the mechanisms of yeast lysis. Microwave, ultrasound or ß-glucanase enzymes are tools that could potentially initiate the release of yeast cell compounds into wine. Further investigation into the impact of such treatments on the flavour and aroma profiles of the wines through sensory evaluation is warranted.

Does the Temperature of the prise de mousse Affect the Effervescence and the Foam of Sparkling Wines?

The persistence of effervescence and foam collar during a Champagne or sparkling wine tasting constitute one, among others, specific consumer preference for these products. Many different factors related to the product or to the tasting conditions might influence their behavior in the glass. However, the underlying factor behind the fizziness of these wines involves a second in-bottle alcoholic fermentation, also well known as the prise de mousse. The aim of this study was to assess whether a low temperature (13 °C) or a high temperature (20 °C) during the in-bottle fermentation might have an impact on the effervescence and the foaming properties (i.e., collar height and bubble size) of three French sparkling wines (a Crémant de Loire and two Champagne wines), under standard tasting conditions. Our results showed that sparkling wines elaborated at 13 °C and served in standard tasting conditions (i.e., 100 mL, 18 °C) had better ability to keep the dissolved CO2 (between 0.09 and 0.30 g/L) in the liquid phase than those elaborated at 20 °C (with P < 0.05). Most interestingly, we also observed, for the Crémant de Loire and for one Champagne wine, that the lower the temperature of the prise de mousse, the smaller (with P < 0.05) the bubbles in the foam collar throughout the wine tasting.

Effect of endogenous CO2 overpressure on the yeast "stressome" during the "prise de mousse" of sparkling wine.

Sparkling wines elaboration by the "Champenoise" method involves a second fermentation of a base wine in hermetically sealed bottles and a subsequent aging period. The whole process is known as "prise de mousse". The endogenous CO2 pressure produced during the second fermentation by the yeast Saccharomyces cerevisiae could modify the sub-proteome involved in the response to different stresses, or "stressome", and cell viability thus affecting the wine organoleptic properties. This study focuses on the stressome evolution along the prise de mousse under CO2 overpressure conditions in an industrial S. cerevisiae strain. The results reveal an important effect of endogenous CO2 overpressure on the stress sub-proteome, cell viability and metabolites such as glycerol, reducing sugars and ethanol. Whereas the content of glycerol biosynthesis-related proteins increased in sealed bottle, those involved in the response to toxic metabolites like ROS, ethanol, acetaldehyde and acetic acid, decreased in content. Proteomic profile obtained in this study may be used to select suitable wine yeast strains for sparkling wine elaboration and improve their stress tolerance.

Chemical compounds and mechanisms involved in the formation and stabilization of foam in sparkling wines.

The visual properties of sparkling wine including foam and bubbles are an indicator of sparkling wine quality. Foam properties, particularly foam height (FH) and foam stability (TS), are significantly influenced by the chemical composition of the wine. This review investigates our current knowledge of specific chemical compounds and, the mechanisms by which they influence the foam properties of sparkling wines. Grape and yeast proteins, amino acids, polysaccharides, phenolic compounds, organic acids, fatty acids, ethanol and sugar are examined with respect to their contribution to foam characteristics in sparkling wines made with the Traditional, Transfer, and Charmat and carbonation methods. Contradictory results have been identified that appear to be due to the analytical methods used to measure and quantify compounds and foam. Biopolymer complexes are discussed and absent knowledge with regards to thaumatin-like proteins (TLPs), polysaccharides, amino acids, oak-derived phenolic compounds and organic acids are identified. Future research is also likely to concentrate on visual analysis of sparkling wines by in-depth imaging analysis and specific sensory analysis techniques.

Immobilisation of yeasts on oak chips or cellulose powder for use in bottle-fermented sparkling wine.

Sparkling wine production comprises two successive fermentations performed by Sacharomyces cerevisiae strains. This research aimed to: develop yeast immobilisation processes on two wine-compatible supports; study the effects of yeast type (IOC 18-2007 and 55A) and the immobilisation support type (oak chips and cellulose powder) on the fermentation kinetics, the deposition rate of lees and the volatile composition of the finished sparkling wine; compare the fermentation parameters of the wines inoculated with immobilised or non-immobilised cells. Proper immobilisation of yeast on oak chips and cellulose powder was demonstrated by electron microscopy. Total sugar consumption occurred in under 60 days in all bottles, regardless of the strain used and the way they were inoculated in wine. Deposition of lees was 3-fold faster in the bottles containing immobilised cells than in those with free cells; no addition of adjuvants was necessary. The analysis of the volatile compounds of the finished sparkling wines showed significant differences in the formation of esters, acids, alcohols, aldehydes and lactones according to the yeast and the immobilisation support used. Oak chips were the more appropriate support for yeast immobilisation. No significant differences in the sensorial analysis of the sparkling wines produced by the different strategies were found.

Effects of different oenological techniques on the elaboration of adequate base wines for red sparkling wine production: phenolic composition, sensory properties and foam parameters.

BACKGROUND: In red sparkling winemaking it is essential to obtain base wines with moderate alcohol content, adequate mouthfeel and color intensity. The aim of this work was to study oenological techniques to obtain adequate base wines for production of red sparkling wine by traditional methods: pre-fermentative cold maceration with dry ice and délestage with premature grapes; and sugar reduction in must and partial dealcoholisation of wine with mature grapes. The effect on oenological parameters, e.g. phenolic content, foam and sensory characteristics, was studied in sparkling wines aged on the lees in bottles for 9 months followed by aging for12 months in bottles after disgorging. RESULTS: Pre-fermentative cold maceration was the only treatment that increased the content of anthocyanins in sparkling wines at both stages of aging. Sparkling wines elaborated using délestage showed the highest mean values of the degree of polymerization of proanthocyanidins. Sparkling wines from mature grapes were given higher valuation in the gustatory phase. Sparkling wines elaborated using pre-fermentative cold maceration were given the highest valuation for foam quality. CONCLUSIONS: Pre-fermentative cold maceration is a viable alternative to common techniques for increasing the anthocyanin content in wines from premature grapes. It would therefore be a good option to obtain adequate base wines. © 2019 Society of Chemical Industry.

The role of production process and information on quality expectations and perceptions of sparkling wines.

BACKGROUND: This study, by combining sensory and experimental economics techniques, aims to analyse to what extent the production process, and the information about it, may affect consumer preferences. Sparkling wines produced by Champenoise and Charmat methods were the object of the study. A quantitative descriptive sensory analysis with a trained panel and non-hypothetical auctions combined with hedonic ratings involving young wine consumers (N = 100), under different information scenarios (Blind, Info and Info Taste), were performed. RESULTS: The findings show that the production process impacts both the sensory profile of sparkling wines and consumer expectations. In particular, the hedonic ratings revealed that when tasting the products, both with no information on the production process (Blind) and with such information (Info Taste), the consumers preferred the Charmat wines. On the contrary, when detailed information on the production methods was given without tasting (Info), consumers liked the two Champenoise wines more. CONCLUSION: It can be concluded that sensory and non-sensory attributes of sparkling wines affect consumers' preferences. Specifically, the study suggests that production process information strongly impacts liking expectations, while not affecting informed liking. © 2018 Society of Chemical Industry.

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

Traditional sparkling wines are produced by the refermentation of a base wine with yeast in the bottle followed by a critical period of aging. During the often lengthy aging process, yeast undergoes cell death and autolysis to release cellular compounds that over time ultimately contribute to the flavor and appearance of the product. While accelerating yeast autolysis for sparkling wine production has been the focus of several studies, employing overexpressed native yeast alleles for this purpose remains poorly explored. Here, we show that the overexpression of native yeast genes, specifically selected autophagic genes, results in accelerated cell death in nitrogen starvation and base wine refermentation. We show ATG3 or ATG4 overexpression has pleiotropic intracellular ramifications including reduced turnover of autophagic cargo, vacuolar fragmentation, abnormal accumulation of lipids, and accelerated accumulation of reactive oxygen species (ROS), all of which precede accelerated cell death. Our findings suggest that the increased expression of autophagy-related genes, such as ATG3 and ATG4, in industrial wine yeast can serve as a suitable marker or breeding strategy to accelerate the cell death and autolysis of wine yeast during sparkling wine production.

Torulaspora delbrueckii for secondary fermentation in sparkling wine production.

In the search for the desired oenological features and flavour complexity of wines, there is growing interest in the potential use of non-Saccharomyces yeast that are naturally present in the winemaking environment. Torulaspora delbrueckii is one such yeast that has seen profitable use in mixed fermentations with Saccharomyces cerevisiae and with different grape varieties. T. delbrueckii can have positive and distinctive impacts on the overall aroma of wines, and has also been used at an industrial level. Here, T. delbrueckii was successfully used in pure and mixed secondary fermentations for sparkling wine. The two selected T. delbrueckii strains used completed the secondary fermentation 'prise de mousse' in these pure and mixed fermentations. The sparkling wines obtained with T. delbrueckii showed different aromatic compositions and sensory profiles to those of S. cerevisiae. T. delbrueckii strain DiSVA 130 showed high esters production and significantly high scores for some of the aromatic descriptors that positively influence the sensory profile of sparkling wine. Thus, the use of T. delbrueckii in pure and mixed fermentations is a suitable strategy to further develop the flavour complexity during secondary fermentation of sparkling wines.

Influence of Grape Berry Maturity on Juice and Base Wine Composition and Foaming Properties of Sparkling Wines from the Champagne Region.

In sparkling wine cool-climate regions like Champagne, it is sometimes necessary to pick the healthy grape clusters that have a relatively low maturity level to avoid the deleterious effects of Botrytis cinerea. In such conditions, we know that classical oenological parameters (sugars, pH, total acidity) may change but there is little information concerning the impact of grape berry maturity on wine proteins and foaming properties. Therefore, healthy grapes (Chardonnay and Pinot meunier) in 2015 and 2016 were picked at different maturity levels within the range of common industrial maturity for potential alcohol content 8⁻11% v/v in the Champagne region. Base wine protein content and foamability, and oenological parameters in grape juice and their corresponding base wines, were investigated. The results showed that base wine protein contents (analyzed by the Bradford method and by electrophoresis) and foamability were higher when the grapes were riper. The Pearson's correlation test found significant positive correlations (r = 0.890⁻0.997, p < 0.05) between Chardonnay grape berry maturity degree (MD) and base wine foamability in both vintages. Strong correlations between MD and most of the oenological parameters in grape juice and base wine were also found for the two cultivars. Under the premise of guaranteed grape health, delaying harvest date is an oenological decision capable of improving base wine protein content and foamability.

Polysaccharides, oligosaccharides and nitrogenous compounds change during the ageing of Tempranillo and Verdejo sparkling wines.

BACKGROUND: Verdejo and Tempranillo are traditional varieties for producing still wines; however, they could provide an alternative for the manufacturing of sparkling wines. Sparkling wines were elaborated by the traditional method, followed by ageing on lees for 9 months. A study on the changes that take place in polysaccharides, oligosaccharides and nitrogenous compounds during the ageing on lees of Tempranillo and Verdejo sparkling wines has been undertaken. RESULTS: Mannoproteins and the glucose residue of oligosaccharides were the major carbohydrates detected in all vinification stages. Yeast polysaccharides and glucan-like structures of the oligosaccharides increased after 3 months of ageing. The evolution of yeast polysaccharides and the composition of PRAG-like structure were different among grape varieties. A decrease in amino acids and biogenic amines was observed during the ageing. The contents of polysaccharides, oligosaccharides and nitrogenous compound were significantly higher in Tempranillo than in Verdejo sparkling wines at the end of the ageing period. CONCLUSION: Polysaccharides and oligosaccharides from yeast were more significant autolysis markers of sparkling wines than the nitrogenous compounds. Our data suggest a potential cultivar effect on the evolution of yeast polysaccharides and on the composition of PRAG, which may influence the physico-chemical and sensory properties of sparkling wines. © 2017 Society of Chemical Industry.

The effect of sucrose addition at dosage stage on the foam attributes of a bottle-fermented English sparkling wine.

BACKGROUND: Approximately two-thirds of wine produced in the UK is bottle-fermented sparkling wine. Effervescence and foamability are key features used to assess English sparkling wine (ESW) quality. A critical, yet understudied, area of research is the potential for dosage to influence foam behaviour via associated changes in wine viscosity. RESULTS: In this study, dosage treatments of five increasing levels of sucrose (from 0 to 31 g L-1 ) were added to an ESW. After storage, the foamability attributes of the wines were analysed via an adapted Mosalux method and a novel image analysis method combined with free pour of the wine. Results indicate that increasing sucrose concentration improved foam formation, but reduced foam stability, likely due to the sucrose added causing a modification in wine viscosity. CONCLUSIONS: These results highlight the impact that dosage treatments can have on the quality of foam produced upon pouring, and therefore have the potential to inform future sparkling winemaking practices. © 2017 Society of Chemical Industry.

Evidence of gushing induction by Penicillium oxalicum proteins.

AIMS: The aim of this study was to elucidate whether grape-associated fungi exert an influence on gushing by their production of surface-active compounds. METHODS AND RESULTS: In preliminary experiments, 58 grape-associated isolates of species within Penicillium and Aspergillus genera were tested for their ability to modify the surface activity of culture supernatants. As the genus Penicillium had a higher potential to change surface activity, further research focused on that genus. Subsequently, supernatants of 36 Penicillium isolates were assessed for their potential to induce gushing in a model system. Isolates of Penicillium oxalicum had the highest potential. Different external factors were investigated for their influence on the intensity of gushing. By using reversed-phase high performance liquid chromatography and subsequent MALDI-TOF MS, SDS-PAGE and nano-ESI-LC-MS/MS analysis, two proteins in the exoproteome of P. oxalicum were identified, which could be linked to the induction of gushing. CONCLUSIONS: Our results suggest that infection of grapes by P. oxalicum may contribute to gushing in sparkling wine. SIGNIFICANCE AND IMPACT OF THE STUDY: In contrast to gushing of beer, the reason for its development in sparkling wine is widely unexplored. Nonetheless, sparkling wine producers have also been affected by this economically damaging phenomenon. This study has first suggested about the occurrence of primary gushing in sparkling wine.

Biodiversity of autolytic ability in flocculent Saccharomyces cerevisiae strains suitable for traditional sparkling wine fermentation.

Yeasts involved in secondary fermentation of traditional sparkling wines should show specific characteristics, such as flocculation capacity and autolysis. Recently it has been postulated that autophagy may contribute to the outcome of autolysis. In this study, 28 flocculent wine Saccahromyces cerevisiae strains characterized by different flocculation degrees were studied for their autolytic and autophagic activities. Autolysis was monitored in synthetic medium through the determination of amino acid nitrogen and total proteins released. At the same time, novel primer sets were developed to determine the expression of the genes ATG1, ATG17 and ATG29. Twelve strains were selected on the basis of their autolytic rate and ATG gene expressions in synthetic medium and were inoculated in a base wine. After 30, 60 and 180 days the autolytic process and ATG gene expressions were evaluated. The obtained data showed that autolysis and ATG gene expressions differed among strains and were independent of the degree of flocculation. This biodiversity could be exploited to select new starter stains to improve sparkling wine production. Copyright © 2016 John Wiley & Sons, Ltd.