Interactions between Starmerella bacillaris and Saccharomyces cerevisiae during sequential fermentations influence the release of yeast mannoproteins and impact the protein stability of an unstable wine.

Wine protein haze formation is a problem due to grape proteins aggregation during wine storage. The cell wall components of wine yeasts, particularly high molecular weight mannoproteins, have a protective effect against haze formation, although their involvement remains poorly understood. This study aimed at characterizing glycosylated proteins released by Starmerella bacillaris and Saccharomyces cerevisiae during single and sequential fermentations in a synthetic must, and testing their impact on wine protein stability. Mannoproteins-rich extracts from sequential fermentations showed an increase in the low MW polysaccharide fraction and, when added to an unstable wine, had a greater effect on protein stability than S. cerevisiae extracts. Shotgun proteomics approaches revealed that the identified cell wall proteins exclusively found in sequential fermentations were produced by both S. bacillaris (MKC7, ENG1) and S. cerevisiae (Bgl2p). Moreover, sequential fermentations significantly increased the expression of Scw4p and 1,3-beta-glucanosyltransferase (GAS5), produced by S. cerevisiae. Finally, some of the key proteins identified might play a positive role in increasing wine protein stability.

Characterization of Mannoprotein Structural Diversity in Wine Yeast Species.

The structure of yeast cell wall (CW) mannoproteins (MPs) influences their impact on wine properties. Yeast species produce a diverse range of MPs, but the link between properties and specific structural features has been ill-characterized. This study compared the protein and polysaccharide moieties of MP-rich preparations from four strains of four different enologically relevant yeast species, named Saccharomyces boulardii (SB62), Saccharomyces cerevisiae (SC01), Metschnikowia fructicola (MF77), and Torulaspora delbrueckii (TD70), and a commercial MP preparation. Monosaccharide determination revealed that SB62 MPs contained the highest mannose/glucose ratio followed by SC01, while polysaccharide size distribution analyses showed maximum molecular weights ranging from 1349 kDa for MF77 to 483 kDa for TD70. Protein identification analysis led to the identification of unique CW proteins in SB62, SC01, and TD70, as well as some proteins shared between different strains. This study reveals MP composition diversity within wine yeasts and paves the way toward their industrial exploitation.

Multivariate prediction of Saliva Precipitation Index for relating selected chemical parameters of red wines to the sensory perception of astringency.

Astringency is an essential sensory attribute of red wine closely related to the saliva precipitation upon contact with the wine. In this study a data matrix of 52 physico-chemical parameters was used to predict the Saliva Precipitation Index (SPI) in 110 Italian mono-varietal red wines using partial least squares regression (PLSr) with variable selection by Variable Importance for Projection (VIP) and the significance of regression coefficients. The final PLSr model, evaluated using a test data set, had 3 components and yielded an R2test of 0.630 and an RMSEtest of 0.994, with 19 independent variables whose regression coefficients were all significant at p < 0.05. Variables selected in the final model according to the decreasing magnitude of their absolute regression coefficient include the following: Procyanidin B1, Epicatechin terminal unit, Total aldehydes, Protein content, Vanillin assay, 520 nm, Polysaccharide content, Epigallocatechin PHL, Tartaric acid, Volatile acidity, Titratable acidity, Catechin terminal unit, Proanthocyanidin assay, pH, Tannin-Fe/Anthocyanin, Buffer capacity, Epigallocatechin PHL gallate, Catechin + epicatechin PHL, and Tannin-Fe. These results can be used to better understand the physico-chemical relationship underlying astringency in red wine.

Effect of Interspecific Yeast Hybrids for Secondary In-Bottle Alcoholic Fermentation of English Sparkling Wines.

In sparkling winemaking, only a few yeast strains are regularly used for the secondary in-bottle alcoholic fermentation (SiBAF). Recently, advances in yeast development programs have yielded new breeds of interspecific wine yeast hybrids that ferment efficiently while producing novel flavors and aromas. In this work, the chemical and sensorial impacts of the use of interspecific yeast hybrids for SiBAF were studied using three commercial English base wines prepared for SiBAF using two commercial and four novel interspecific hybrids. After 12 months of lees aging, the chemical and macromolecular composition, phenolic profile, foaming, viscosity and sensory properties of the resulting 13 wines were assessed. Chemically, the yeast strains did not result in significant differences in the main wine parameters, while some differences in their macromolecular contents and sensory characteristics were noticeable. The foamability was mostly unaffected by the strain used; however, some effect on the foam stability was noticeable, likely due to the differences in polysaccharides released into the wines by the yeast strains. The wines exhibited different sensory characteristics in terms of aroma and bouquet, balance, finish, overall liking and preference, but these were mostly attributable to the differences in the base wines rather than the strain used for SiBAF. Novel interspecific yeast hybrids can be used for the elaboration of sparkling wines, as they provided wines with chemical characteristics, flavor and aroma attributes similar to those of commonly used commercial Saccharomyces cerevisiae strains.

The contribution of varietal thiols in the diverse aroma of Italian monovarietal white wines.

Thanks to their low odor detection thresholds, free varietal thiols (VTs) play a key role in the primary aroma of wines, to which they confer an intense scent reminiscent of box tree, grapefruit, citrus fruits, passionfruit and cat urine odor. Excluding wines from a few VT-rich grapevine cultivars, VTs appear to be present in most cultivars at trace levels, although a comprehensive dataset is still missing. The low concentration of VTs combined with their high reactivity and matrix complexity make their determination in wines a challenging task. In this research an optimized liquid chromatography - tandem mass spectrometry (LC-MS/MS) method was validated and used for the quantification of 4-methyl-4-sulfanylpentan-2-one (4-MSP), 3-sulfanylhexan-1-ol (3-SH), 3-sulfanylhexyl acetate (3-SHA) and ethyl 3-sulfanylpropionate (E3SP) in 246 samples (vintage 2019) representative of 18 monovarietal Italian white wines. VTs were detected in all cultivars even though higher values of 3-SH were found in Lugana, Müller-Thurgau and Verdicchio cultivars. Müller-Thurgau wines showed the highest level of 4-MSP, that was mainly correlated to the odor descriptors of passionfruit and box tree/cat urine. The VTs composition of Müller-Thurgau was confirmed on a second set of 50 wines from different vintages. From a sensory perspective, the samples of Müller-Thurgau showed the best positive correlations between chemical variables and the odor descriptors thiol note, passion fruit and box tree/cat urine. These notes are significantly related to 4-MSP, suggesting that it could play a relevant olfactory role for the aroma of Müller-Thurgau wines. Sorting analysis allowed to group these wines according to their thiolic characteristics. The chemical variables and the odor descriptors attributable to the thiol notes are important for Müller-Thurgau and Lugana wines, while the contribution of thiol notes was sensorially negligible for the other wines.

Emulsifying activity of potato proteins in the presence of k-carrageenan at different pH conditions.

Oil in Water (3:1) emulsions were prepared using potato proteins in the presence or absence of 0.2% k-carrageenan at different pH conditions (3.0, 7.0, and 4.8). These emulsions showed different droplet sizes, stability, appearance, and rheological properties. The best emulsion stability was achieved combining potato proteins and k-carrageenan at pH 3.0, where uniform and small oil droplets (30 µm) were observed. The rheological properties of the emulsions were also different. The highest viscosity and G' were shown by the emulsion prepared with the addition of k-carrageenan at pH 3.0, this being attributed to the onset of a gel-like viscoelastic structure in these conditions. SDS-PAGE indicated that the superior properties of the emulsion prepared with k-carrageenan at pH 3.0 can be attributed to an electrostatic interaction between the positively charged potato proteins and the anionic polysaccharide. This interaction allowed the formation of a strong molecular network able to stabilize the system.

Diversity of Italian red wines: A study by enological parameters, color, and phenolic indices.

An extensive survey was conducted on 110 Italian monovarietal red wines from a single vintage to determine their standard compositional, color, and phenolic characteristics, analysing more than 35 parameters evaluated through methods commonly used in the wine industry. 'Primitivo' achieved the highest average alcohol strength (15.4% v/v) and dry extract values, while 'Cannonau' showed the lowest total acidity. 'Corvina' had the lowest phenolic content (1065 mg/L by Folin-Ciocalteu assay), remarkably different from the highest found in 'Sagrantino' (3578 mg/L), the latter being also the richest variety in both proanthocyanidins and vanillin-reactive flavanols. 'Teroldego' wines were the richest in both total and monomeric anthocyanins (702 and 315 mg/L, respectively), followed by 'Aglianico' and 'Raboso Piave', while 'Corvina', 'Nebbiolo', and 'Nerello Mascalese' were the poorest. 'Montepulciano' and 'Sangiovese' showed intermediate values for the majority of the parameters analyzed. A multivariate PCA-DA approach allowed achieving both a classification of the different wines as well as the discrimination of 'Sangiovese' wines produced in two regions (Emilia Romagna and Toscana) that returned a 42-66% success rate depending on the zone considered. Taking into account the number and diversity of the wines analyzed, a correlation study helped in better understanding the underlying relations between the most common and widespread analytical techniques for phenolic and color determinations.

Techniques for Mitigating the Effects of Smoke Taint While Maintaining Quality in Wine Production: A Review.

Smoke taint has become a prominent issue for the global wine industry as climate change continues to impact the length and extremity of fire seasons around the world. Although the issue has prompted a surge in research on the subject in recent years, no singular solution has yet been identified that is capable of maintaining the quality of wine made from smoke-affected grapes. In this review, we summarize the main research on smoke taint, the key discoveries, as well as the prevailing uncertainties. We also examine methods for mitigating smoke taint in the vineyard, in the winery, and post production. We assess the effectiveness of remediation methods (proposed and actual) based on available research. Our findings are in agreement with previous studies, suggesting that the most viable remedies for smoke taint are still the commercially available activated carbon fining and reverse osmosis treatments, but that the quality of the final treated wines is fundamentally dependent on the initial severity of the taint. In this review, suggestions for future studies are introduced for improving our understanding of methods that have thus far only been preliminarily investigated. We select regions that have already been subjected to severe wildfires, and therefore subjected to smoke taint (particularly Australia and California) as a case study to inform other wine-producing countries that will likely be impacted in the future and suggest specific data collection and policy implementation actions that should be taken, even in countries that have not yet been impacted by smoke taint. Ultimately, we streamline the available information on the topic of smoke taint, apply it to a global perspective that considers the various stakeholders involved, and provide a launching point for further research on the topic.

Optimised Extraction and Preliminary Characterisation of Mannoproteins from Non-Saccharomyces Wine Yeasts.

The exogenous application of yeast-derived mannoproteins presents many opportunities for the improvement of wine technological and oenological properties. Their isolation from the cell wall of Saccharomycescerevisiae has been well studied. However, investigations into the efficiency of extraction methods from non-Saccharomyces yeasts are necessary to explore the heterogeneity in structure and composition that varies between yeast species, which may influence wine properties such as clarity and mouthfeel. In this study, nine yeast strains were screened for cell wall mannoprotein content using fluorescence microscopy techniques. Four species were subsequently exposed to a combination of mechanical and enzymatic extraction methods to optimize mannoprotein yield. Yeast cells subjected to 4 min of ultrasound treatment applied at 80% of the maximum possible amplitude with a 50% duty cycle, followed by an enzymatic treatment of 4000 U lyticase per g dry cells weight, showed the highest mannoprotein-rich yield from all species. Furthermore, preliminary evaluation of the obtained extracts revealed differences in carbohydrate/protein ratios between species and with increased enzyme incubation time. The results obtained in this study form an important step towards further characterization of extraction treatment impact and yeast species effect on the isolated mannoproteins, and their subsequent influence on wine properties.

Current and future strategies for wine yeast lees valorization.

Wine lees is a sludge material mainly composed of dead yeast precipitated at the bottom of wine tanks. Along with grape pomace and grape stalks, it is one of the main by-products of the winemaking industry. Given that wine lees are considered a soil pollutant, their disposal represents a cost for wineries. Numerous wine lees recovery and valorization strategies have been proposed, with a particularly steep increase in published research in recent years. This attention is strictly linked to the concepts of circular economy and environmental sustainability that are attracting the interest of the scientific community. In this review, an overview on the available wine lees recovery and valorization strategies is reported. Additionally, the methods for the extraction and valorization of yeast's cell wall polysaccharides (ß-glucans and mannoproteins) are discussed. Finally, current and future innovative applications in different sectors of yeast ß-glucans and mannoproteins are described and critically discussed.

Evaluation of the phenolic profile and immunoreactivity of Mal d 3 allergen in ancient apple cultivars from Italy.

BACKGROUND: Since the second half of the 20th century, the cultivation of ancient and local apple cultivars has almost disappeared from orchards in Italy. Some of these ancient apple cultivars often possess high nutraceutical values and display lower allergenicity than the modern ones, supporting the so-called 'green revolution' theory. RESULTS: In this study, the phenolic composition and the antioxidant activity of five ancient apple cultivars ('Belfiore', 'Pomella Genovese', 'Gravenstein', 'Bella del Bosco', and 'Piatlin') were compared with a 'Golden Delicious' commercial cultivar. Additionally, apples were tested for their potential allergenicity by detecting the presence of Mal d 3, a non-specific lipid transfer protein that represents the main apples' allergen. All apples came from northern Italy (Trentino Region) and were organically produced. Results showed that, for all cultivars, the skins contained more polyphenols than the pulps. 'Bella del Bosco' had the highest amount of polyphenols and antioxidant activity, whereas 'Piatlin' had the lowest phenolic content. All ancient cultivars presented a higher amount of pulp phenolic compounds than 'Golden Delicious'. Immunoblotting techniques showed that 'Bella del Bosco' and 'Piatlin' had very low quantities of Mal d 3 allergen; hence, they can be considered hypoallergenic cultivars. CONCLUSIONS: The preservation of ancient apple cultivars would be of great importance, not only to maintain the biodiversity but also for their nutritional properties. The hypoallergenic activity of some of these cultivars could be of interest also for the preparation of different apple-based products. © 2020 Society of Chemical Industry.

Use of Untargeted Liquid Chromatography-Mass Spectrometry Metabolome To Discriminate Italian Monovarietal Red Wines, Produced in Their Different Terroirs.

The aim of this project was to register, in a liquid chromatography-mass spectrometry-based untargeted single-batch analysis, the metabolome of 11 single-cultivar, single-vintage Italian red wines (Aglianico, Cannonau, Corvina, Montepulciano, Nebbiolo, Nerello, Primitivo, Raboso, Sagrantino, Sangiovese, and Teroldego) from 12 regions across Italy, each one produced in their terroirs under ad hoc legal frameworks to guarantee their quality and origin. The data provided indications regarding the similarity between the cultivars and highlighted a rich list of putative biomarkers of origin wines (pBOWs) characterizing each individual cultivar-terroir combination, where Primitivo, Teroldego, and Nebbiolo had the maximum number of unique pBOWs. The pBOWs included anthocyanins (Teroldego), flavanols (Aglianico, Sangiovese, Nerello, and Nebbiolo), amino acids and N-containing metabolites (Primitivo), hydroxycinnamates (Cannonau), and flavonols (Sangiovese). The raw data generated in this study are publicly available and, therefore, accessible and reusable as a baseline data set for future investigations.

The Secondary Structure of a Major Wine Protein is Modified upon Interaction with Polyphenols.

Polyphenols are an important constituent of wines and they are largely studied due to their antioxidant properties and for their effects on wine quality and stability, which is also related to their capacity to bind to proteins. The effects of some selected polyphenols, including procyanidins B1 and B2, tannic acid, quercetin, and rutin, as well as those of a total white wine procyanidin extract on the conformational properties of the major wine protein VVTL1 (Vitis vinifera Thaumatin-Like-1) were investigated by Synchrotron Radiation Circular Dichroism (SRCD). Results showed that VVTL1 interacts with polyphenols as demonstrated by the changes in the secondary (far-UV) and tertiary (near-UV) structures, which were differently affected by different polyphenols. Additionally, polyphenols modified the two melting temperatures (TM) that were found for VVTL1 (32.2 °C and 53.9 °C for the protein alone). The circular dichroism (CD) spectra in the near-UV region revealed an involvement of the aromatic side-chains of the protein in the interaction with phenolics. The data demonstrate the existence of an interaction between polyphenols and VVTL1, which results in modification of its thermal and UV denaturation pattern. This information can be useful in understanding the behavior of wine proteins in presence of polyphenols, thus giving new insights on the phenomena that are involved in wine stability.

Wine Fining with Plant Proteins.

Fining treatments involve the addition of a substance or a mixture to wine, and are generally carried out in order to clarify, stabilize or modify the wine's organoleptic characteristics. Usually these fining agents will bind the target compound(s) to form insoluble aggregates that are subsequently removed from the wine. The main reasons to perform wine fining treatments are to carry out wine clarification, stabilization and to remove phenolic compounds imparting unwanted sensory characteristics on the wine, which is an operation that often relies on the use of animal proteins, such as casein, gelatin, egg and fish proteins. However, due to the allergenic potential of these animal proteins, there is an increasing interest in developing alternative solutions including the use of fining proteins extracted from plants (e.g., proteins from cereals, grape seeds, potatoes, legumes, etc.), and non-proteinaceous plant-based substances (e.g., cell wall polysaccharides and pomace materials). In this article, the state of the art alternative fining agents of plant origins are reviewed for the first time, including considerations of their organoleptic and technological effects on wine, and of the allergenic risks that they can pose for consumers.

Characterization and emulsifying properties of extracts obtained by physical and enzymatic methods from an oenological yeast strain.

BACKGROUND: Glycosylated compounds are one of the main fractions of the yeast cell wall. Thanks to their amphiphilic structure, they have been studied as stabilizers in food emulsions over a broad range of pH conditions with encouraging results. Nevertheless, extraction costs still represent an important limit for their application in the food industry. RESULTS: In this research, four extraction methods were applied to yeast cells exploiting both physical (heating and sonication) and enzymatic approaches (use of three industrial enzyme preparations, namely Glucanex®, Sur Lies and Elevage). A fifth method involving a pure ß-glucanase enzyme (Zymolyase) was taken as reference. These extraction methods were applied to the oenological strain Saccharomyces cerevisiae EC1118, and their extraction yields and chemical properties (quantitative and qualitative determination of sugars and proteins) were studied. Emulsifying activities were determined at three different pH values (3, 5 and 7). Extractions with Physical, Glucanex and Sur Lies methods were the most successful approaches to obtain relevant amounts of yeast compounds with good emulsifying activities for 2:1 oil-in-water emulsions at pH 3 and 7 over 48 h. CONCLUSIONS: These results indicate that there is the potential for the extraction approaches here proposed to become viable tools for the recovery of yeast compounds to be used as emulsifiers in foods. This approach can be considered as the starting point to explore the possibility to exploit yeast by-products from the fermentation processes (e.g. fermentation lees from wine and beer making) as valuable compounds for food applications. © 2019 Society of Chemical Industry.

Sensory characterization of cucumbers pickled with verjuice as novel acidifying agent.

Verjuice is an acidic unfermented unripe grape juice that has recently been proposed as an alternative to vinegar in different food preparations. In this study, pickled cucumber preserves were industrially prepared using two dilutions of verjuice as acidifying agent and compared with the traditional vinegar-pickled samples. Samples were chemically characterized and subjected to sensory analyses using a trained panel and consumers. The different acidifying agents resulted in pickled cucumbers giving different olfactory and gustatory evaluations. No differences in appearance and texture were found. Despite showing chemical and sensory differences, pickles preserved with verjuice and vinegar had similar overall liking scores for visual, olfactory and gustatory aspects. A key feature of pickles prepared with verjuice is their lack of acetic acid potentially conferring them an additional value, suggesting that verjuice represents a viable alternative to vinegar for the production of pickles with distinctive sensorial properties.

Influence of the reducing environment in the misfolding of wine proteins.

While proteins are present in wine at low concentration, and are largely associated with undesirable haze formation in white wines, certain types or fractions make direct and indirect contributions to sensory quality and physical stability. The proteins found in wine represent a small subclass of the total pool of grape proteins that remain soluble in the non-physiological conditions of the wine matrix which is characterised by the presence of alcohol, high acidity, and relatively high levels of phenolic compounds. Although initially stable in these conditions, during storage of white and rosé wines proteins undergo changes leading to haze formation which is considered one of the most relevant non-microbiological defects, and which makes the wine commercially unacceptable. This phenomenon involves the two most abundant proteins present in wines: thaumatin-like proteins and chitinases, both belonging to pathogenesis-related proteins of the grape berry. Haze formation is often triggered by thermal fluctuations occurring during storage of white wines, although the presence of other non-protein-related factors seems to be necessary. Here, we review the characteristics of these two protein families and the factors that influence their solubility with a focus on the disulfide bonds reduction as a possible trigger for the onset of their aggregation.

A Novel Method for the Quantification of White Wine Mannoproteins by a Competitive Indirect Enzyme-Linked Lectin Sorbent Assay (CI-ELLSA).

Mannoproteins (MPs) are cell wall proteoglycans released in wine by yeast during fermentation and ageing on lees, a procedure used for the production of several wines to enrich them in these components with consequences from both a technological and sensory point of view. Given the significance that wine MPs have for wine quality, winemakers would welcome a simple and accurate method for their quantification, as this would allow them to have a better control of this aspect at different winemaking stages. This study develops and validates a novel, simple and accurate method for MPs quantification in white wines based on a competitive indirect enzyme-linked lectin sorbent assay (CI-ELLSA), using the highly mannosylated yeast invertase as the standard. The method utilizes the lectin concanavalin A (ConA) as the immobilized ligand for MPs, and peroxidase, an enzyme rich in mannose, as the competitor for ConA. After addition of the peroxidase substrate, the intensity of the signal produced by the activity of this enzyme (absorbance at 450 nm) is inversely proportional to the amount of mannosylated proteins in the sample. Results have been validated on several wine styles including still, sparkling and sweet wines.

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.