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…

Water deficit differentially impacts the performances and the accumulation of grape metabolites of new varieties tolerant to fungi

The use of resistant varieties is a long-term but promising solution to reduce chemical input in viticulture. Several important breeding programs in Europe and abroad are now releasing a range of new hybrids performing well regarding fungi susceptibility and producing good quality wines. Unfortunately, insufficient attention is paid by the breeders to the adaptation of these varieties to climatic changes, notably to the increased climatic demand and water deficit (WD). Thus, prior to the adoption of such varieties by the wine industry in Mediterranean regions, there is a need to consider their suitability to WD. This study aimed to characterize the different drought-strategies adopted by 6 new resistant varieties selected by INRAE in comparison to Syrah. To allow the assessment of long-term impacts of WD, field-grown vines were exposed to contrasted WD from 2018 to 2021 under a semi-arid Mediterranean climate. A gradient of WD was applied in the field and controlled through plant measurements at the single plant level. Grape development was non-destructively monitored to determine the arrest of berry phloem unloading. The impacts of WD on berry composition, including water, primary metabolites (sugars, organic acids), secondary metabolites (anthocyanins, thiols precursors) and main cations contents, were assessed at this specific stage. Results showed different varietal responses during the year and inter-annual acclimation in terms of plant water use efficiency, biomass accumulation, as well as yield components and berry composition. WD differentially reduced the accumulation of primary metabolites at plant and berry levels, but it little changed their concentrations in the fruits at the ripe stage. Moreover, WD differentially impacted the accumulation of secondary metabolites and major cations between the varieties. In the talk, we’ll present the main results regarding the WD impacts on fruit metabolites and enlarge the reflection about the practical assessment of the grapevine acclimation to WD.

Climate, Viticulture, and Wine … my how things have changed!

The planet is warmer than at any time in our recorded past and increasing greenhouse emissions and persistence in the climate system means that continued warming is highly likely. Climate change has already altered the basic framework of growing grapes for wine production worldwide and will likely continue to do so for years to come. The wine sector can continue to play an important role in leading the agricultural sector in addressing climate change. From developing on…

Impact of long term agroecological and conventional practices on subsurface soil microbiota in Macabeu and Xarel·lo vineyards

There is a growing trend on the transition from conventional to agroecological management of vineyards. However, the impact of practices, such as reduced-tillage, organic fertilization and cover crops, is not well-understood regarding the soil microbial diversity, and its relationship with the soil physicochemical properties in the subsurface depth near the rooting zone. Soil bacterial diversity is an important contributor towards plant health, productivity and response to environmental stresses. A field experiment was conducted by sampling subsurface soil bacterial community (NGS and qPCR) near to the root zone of Macabeu and Xarel·lo vineyards, located at the Penedes. 3 organic (ECO) and 3 conventional (CON) vineyards, with more than 10 years of respective management were sampled (n=5 each plot). ECO practices did not affect bacterial and fungal abundance but increased significantly the ammonium oxidizing bacteria and alpha-diversity (Inv.Simpson). Interestingly beta-diversity was significantly affected by the management strategy. ANOSIM-tests revealed a significative effect of the management (ecological vs conventional) and plot, on the soil microbial structure (ASV abundance). Main phyla depicted were Proteobacteria, Actinobacteria and Acidobacteria, whose relative abundances were not affected by the management. EdgeR assay revealed a significant increase of Cyanobacteria and decrease of Gemmatimonadetes and Firmicutes phyla in ECO. Interestingly, the grapevine variety was not correlated with the soil microbial community structure. Mantel-test revealed an important correlation (Spearman) of some physicochemical parameters with the soil microbiota structure, in order of importance: texture, EC, pH Ca/Mg, Mg/P, K+, Mg2+, Ca2+, SO42-, and OM. N-NH4 and NTK, which were higher in the ECO managed soils, did not correlated significantly with the soil microbiome population. The results revealed the importance of combining a deep physicochemical characterization of each replicate with the microbial diversity assessment to gain better insights on the relationship between soil microbiome and vineyard management.

Phenolic composition of Tempranillo Blanco grapes changes after foliar application of urea

Our research aimed to determine the effect and efficiency of foliar application of urea on the phenolic composition of Tempranillo Blanco grapes. The field experiment was carried out in 2019 and 2020 seasons and the plot was located in D.O.Ca Rioja (North of Spain). The vineyard was Vitis vinifera L. Tempranillo Blanco and grafted on Richter-110 rootstock. The treatments were control (C), whose plants were sprayed with water and three doses of urea: plants were sprayed with urea 3 kg N/ha (U3), 6 kg N/ha (U6) and 9 kg N/ha (U9). The applications were performed in two phenological stages, pre-veraison (Pre) and veraison (Ver). Also, each of the treatments was repeated one week later. Control and treatments were performed in triplicate and arranged in a randomised block design. Grapes were harvested at optimum ripening stage. High-performance liquid chromatography was used to analyse the phenolic composition of the grapes. Finally, the results obtained from the analytical determinations – flavonols, flavanols and non-flavonoid (hydroxybenzoic acids, hydroxycinnamic acids and stilbenes) – were studied statistically by analysis of variance. The results showed that, in 2019, U6-Pre and U9-Pre treatments increased the hydroxybenzoic acid content in grapes, and also all foliar treatments applied at Pre enhanced the stilbene concentration. Moreover, U3-Ver was the only treatment that rose flavonol and stilbene contents in the Tempranillo Blanco grapes. In 2020, all treatments applied at Pre enhanced the flavonol concentration in grapes. Furthermore, U3-Pre and U9-Pre treatments increased stilbene content in grapes. Nevertheless, the hydroxybenzoic acid content was improved by U6-Ver and U9-Ver and besides, hydroxycinnamic acid concentration in grapes was increased by all treatments applied at Ver. In conclusion, the lower and highest dose of urea (U3 and U9), applied at pre-veraison, were the best treatments to improve the Tempranillo Blanco grape phenolic composition.

Spatial variability of temperature is linked to grape composition variability in the Saint-Emilion winegrowing area

Elevated temperature during the grape maturation period is a major threat for grape quality and thus wine quality. Therefore, characterizing the grape composition response to temperature at a larger scale would represent a crucial step towards adaptation to climate change. In response to changes in temperature, various physiological mechanisms regulate grape composition. Primary and secondary metabolisms are both involved in this response, with well-known effects, for example on anthocyanins, and lesser known effects, for example on aromas or aroma precursors. At the field scale or at the regional scale, however, numerous environmental or plant-specific factors intervene to make the effects of temperature difficult to distinguish from overall variability. In this study, it was attempted to overcome this difficulty by selecting well-characterized situations with differing temperatures.
A long-term study of air temperature variability across several Merlot vineyards in the Saint-Emilion and Pomerol wine producing area found significant temperature differences and gradients at various time scales linked to environmental factors. From this study area, a few sites were selected with similar age, soil and training system conditions, and with repeated and contrasted temperature differences during the maturation period. The average temperature difference during the maturation period was about 2°C between cooler and warmer sites, a difference similar to that expected under future climate change scenarios. In close vicinity to the temperature sensors at each site, grape berries were sampled at different times until full maturity during 2019 and 2020. Also, berries from bunches on either side of the row were analyzed separately, allowing an investigation of bunch exposure effect associated with the coupling of berry temperature and solar radiation. Four replicates of pooled berries for each time – site – bunch exposure combination were obtained and analyzed for biochemical composition. Analyses of variance of the biochemical composition data collected at different sampling times reveal significant effects associated with temperature, site, and bunch azimuth. For instance, anthocyanins in grape skins are clearly influenced by temperature and solar radiation exposure, with up to 30% reduction in warmer conditions.