IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Effect of redox mediators on the activity of laccase from Botrytis cinerea against volatile phenols

Effect of redox mediators on the activity of laccase from Botrytis cinerea against volatile phenols

Abstract

Volatile phenols namely 4-ethylphenol and 4-ethylguaiacol are formed by enzymatic decarboxylation of hydroxycinnamic acids by Brettanomyces yeasts to give vinylphenols and subsequent reduction of the vinyl group to form the correspondent ethylphenols. The presence of these compounds in wine affects negatively its aromatic quality, conferring unpleasant animal and phenolic odor when present in quantities above the olfactory detection threshold [1]. Several methods have been described to remove these undesirable compounds from wines, including the use laccase enzymes [2, 3]. Due to this, the aim of this work was to evaluate the effect of several natural redox mediators on the activity of Botrytis cinerea laccase against these volatile phenols.

The ability of Botrytis cinerea laccase to degrade 4-ethylphenol and 4-ethylguaiacol was studied by incubation with the enzyme in acetate buffer and model wine, and several phenolic compounds were individually assayed as mediators. Quantification of volatile phenols was accomplished by GC-MS analysis.

The only use of the Botrytis cinerea laccase was not effective in reducing or removing these off-flavors and the presence of mediators was required under these conditions. All phenolic compounds tested (caftaric acid, quercetin-3-O-rutinoside, catechin, epicatechin, ferulic acid and quercetin) favored the degradation of volatile phenols, achieving higher 4-ethylguaiacol removal percentages than that for 4-ethylphenol. These preliminary results confirm the activity of this type of enzyme against volatile phenols and provide knowledge on the effects of natural mediators on the biodegradation effectiveness of undesirable substances which may alter the quality of wine.

References

1. Petrozziello M, Asproudi A, Guaita M, Borsa D, Motta S, Panero L, Bosso A. 2014. Influence of the matrix composition on the volatility and sensory perception of 4-ethylphenol and 4-ethylguaiacol in model wine solutions. Food Chemistry 149: 197–202.
2. Lustrato G, De Leonardis A, Macciola V, Ranalli G. 2015. Preliminary lab scale of advanced techniques as new tools to reduce ethylphenols content in synthetic wine. Agro FOOD Industry Hi Tech 26:51-54.
3. Moeder M, Martin C, Koeller G. 2004. Degradation of hydroxylated compounds using laccase and horseradish peroxidase immobilized on microporous polypropylene hollow fiber membranes. Journal of Membrane Science 245:183-190.

DOI:

Publication date: June 27, 2022

Issue: IVAS 2022

Type: Poster

Authors

Pérez-Navarro José1,2, Osorio Alises María3, Paniagua Martínez Tania3, Giménez Pol4, Canals Joan Miquel4, Zamora Fernando4, Sánchez-Palomo Eva3, González-Vinas Miguel Ángel3 and Gómez-Alonso Sergio2,3

1Higher Technical School of Agronomic Engineering, University of Castilla-La Mancha.
2Regional Institute for Applied Scientific Research (IRICA), University of Castilla-La Mancha
3Faculty of Chemical Sciences and Technologies, University of Castilla-La Mancha
4Faculty of Oenology, Rovira i Virgili University

Contact the author

Keywords

4-ethylphenol, 4-ethylguaiacol, enzyme, phenolic compounds, fungi

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

Second pruning as a strategy to delay maturation in cv. ‘Touriga nacional’ in the Portuguese Douro region

The advance in maturation of wine grapes is an important climate change risk related effect that could affect warm regions like Portuguese Douro Wine Region. Indeed, the climate analysis over the past years registered a decrease in the precipitation, significant higher average temperatures, and a more frequent occurrence of extreme weather events, including heat waves. In these conditions the length from anthesis until maturation is shortened and the uncoupling of technical and phenolic maturity results in berries with higher sugar concentration (and lower acidity), but lower anthocyanins, tannins, and total phenolic concentration, which produce unbalanced wines. In this work, an innovative strategy of crop forcing, based on forcing vine regrowth after a second pruning of green shoots, was tested, aimed at delaying ripening until the temperature becomes lower and, therefore, preventing acidity loss and increasing anthocyanin-to-sugar ratio. The experiments were conducted in 2019 and 2020 in a commercial vineyard of ‘Touriga Nacional’ located in the Douro Region. Crop forcing was conducted 15 (CF1) to 30 (CF2) days after fruit set. Vines pruned with conventional methods were used as control (CF0). Results confirmed that fruit ripening was shifted from the hot season (August/September), until a cooler period (October through early-November). At harvest, grapevine berries from CF1 and CF2 presented lower pH and higher acidity, than control, with no significant differences in colour intensity and phenolic levels composition. Sugar content was lower in CF2-treated vines in both seasons. However, in CF-treated vines the number and size of clusters were significantly lower (up to 88% reduction) than in control plants. A metabolomics analysis of mature berries from CF-treated vines and control is underway. Crop forcing was indeed effective in producing a more balance berry composition but severely reduced grapevine yield,

What are the optimal ranges and thresholds for berry solar radiation for flavonoid biosynthesis?

In wine grape production, canopy management practices are applied to control the source-sink balance and improve the cluster microclimate to enhance berry composition. The aim of this study was to identify the optimal ranges of berry solar radiation exposure (exposure) for upregulation of flavonoid biosynthesis and thresholds for their degradation, to evaluate how canopy management practices such as leaf removal, shoot thinning, and a combination of both affect the grapevine (Vitis vinifera L. cv. Cabernet Sauvignon) yield components, berry composition, and flavonoid profile under context of climate change. First experiment assessed changes in the grape flavonoid content driven by four degrees of exposure. In the second experiment, individual grape berries subjected to different exposures were collected from two cultivars (Cabernet Sauvignon and Petit Verdot). The third experiment consisted of an experiment with three canopy management treatments (i) LR (removal of 5 to 6 basal leaves), (ii) ST (thinned to 24 shoots per vine), and (iii) LRST (a combination of LR and ST) and an untreated control (UNT). Berry composition, flavonoid content and profiles, and 3-isobutyl 2-methoxypyrazine were monitored during berry ripening. Although increasing canopy porosity through canopy management practices can be helpful for other purposes, this may not be the case of flavonoid compounds when a certain proportion of kaempferol was achieved. Our results revealed different sensitivities to degradation within the flavonoid groups, flavonols being the only monitored group that was upregulated by solar radiation. Within different canopy management practices, the main effects were due to the ST. Under environmental conditions given in this trial, ST and LRST hastened fruit maturity; however, a clear improvement of the flavonoid compounds (i.e., greater anthocyanin) was not observed at harvest. Methoxypyrazine berry content decreased with canopy management practices studied. Although some berry traits were improved (i.e. 2.5° Brix increase in berry total soluble solids) due to canopy management practices (ST), this resulted in a four-fold increase in labor operations cost, two-fold decrease in yield with a 10-fold increase in anthocyanin production cost per hectare that should be assessed together as the climate continues to get hot.

Effects of graft quality on growth and grapevine-water relations

Climate change is challenging viticulture worldwide compromising its sustainability due to warmer temperatures and the increased frequency of extreme events. Grafting Vitis vinifera L.

1H-NMR-based Metabolomics to assess the impact of soil type on the chemical composition of Mediterranean red wines

The aim of this study was to evaluate the effects of different soil types on the chemical composition of Mediterranean red wines, through untargeted and targeted 1H-NMR metabolomics. One milliliter of raw wine was analyzed by means of a Bruker Avance II 400 spectrometer operating at 400.15 MHz. The spectra were recorded by applying the NOESYGPPS1D pulse sequency, to achieve water and ethanol signals suppression. No modification of the pH was performed to avoid any chemical alteration of the matrix. The generation of input variables for untargeted analysis was done via bucketing the spectra. The resulting dataset was preprocessed prior to perform unsupervised PCA, by means of MetaboAnalyst web-based tool suite. The identification of compounds for the targeted analysis was performed by comparison to pure compounds spectra by means of SMA plug-in of MNova 14.2.3 software. The dataset containing the concentrations (%) of identified compounds was subjected to one-way analysis of variance (ANOVA) to highlight significant differences among the wines. The untargeted analysis, carried out through the PCA, revealed a clear differentiation among the wines. The fragments of the spectra contributing mostly to the separation were attributed to flavonoids, aroma compounds and amino acids. The targeted analysis leaded to the identification of 68 compounds, whose concentrations were significant different among the wines. The results were related to soils physical-chemical analysis and showed that: 1) high concentrations of flavan-3-ols and flavonols are correlated with high clay content in soils; 2) high concentrations of anthocyanins, amino acids, and aroma compounds are correlated with neutral and moderately alkaline soil pH; 3) low concentrations of flavonoids and aroma compounds are correlated with high soil organic matter content and acidic pH. The 1H-NMR metabolomic analysis proved to be an excellent tool to discriminate between wines originating from grapes grown on different soil types and revealed that soils in the Mediterranean area exert a strong impact on the chemical composition of the wines.

Climate projections over France wine-growing region and its potential impact on phenology

Climate change represents a major challenge for the French wine industry. Climatic conditions in French vineyards have already changed and will continue to evolve. One of the notable effects on grapevine is the advancing growing season. The aim of this study is to characterise the evolution of agroclimatic indicators (Huglin index, number of hot days, mean temperature, cumulative rainfall and number of rainy days during the growing season) at French wine-growing regions scale between 1980 and 2019 using gridded data (8 km resolution, SAFRAN) and for the middle of the 21th century (2046-2065) with 21 GCMs statistically debiased and downscaled at 8 km. A set of three phenological models were used to simulate the budburst (BRIN, Smoothed-Utah), flowering, veraison and theoretical maturity (GFV and GSR) stages for two grape varieties (Chardonnay and Cabernet-Sauvignon) over the whole period studied. All the French wine-growing regions show an increase in both temperatures during the growing season and Huglin index. This increase is accompanied by an advance in the simulated flowering (+3 to +9 days), veraison (+6 to +13 days) and theoretical maturity (+6 to +16 days) stages, which are more noticeable in the north-eastern part of France. The climate projections unanimously show, for all the GCMs considered, a clear increase in the Huglin index (+662 to 771 °C.days compared to the 1980-1999 period) and in the number of hot days (+5.6 to 22.6 days) in all the wine regions studied. Regarding rainfall, the expected evolution remains very uncertain due to the heterogeneity of the climates simulated by the 21 models. Only 4 regions out of 21 have a significant decrease in the number of rainy days during the growing season. The two budburst models show a strong divergence in the evolution of this stage with an average difference of 18 days between the two models on all grapevine regions. The theoretical maturity is the most impacted stage with a potential advance between 40 and 23 days according to wine-growing regions.