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…

Diagnosis of soil quality and evaluation of the impact of viticultural practices on soil biodiversity in a vineyard in southwestern France

Viticulture is facing two major changes – climate change and agroecological transition. In both cases, soil quality is seen as a lever to move towards a more sustainable viticulture. However, soil biological quality is little considered in the implementation of viticultural practices. Gascogn’Innov (2017-2022) is an Operational Group funded by the European Innovation Partnership for Agriculture. As such, it brings together winegrowers from the south-west of France, scientists, advisors and technicians, around a project focused on viticultural soil biological functioning and the design of technical routes more respectful toward soil heritage. To achieve this, the project aims to acquire references on the impact of viticultural practices on soil biology from a dynamic way, and to test a methodology to integrate information provided by the soil bioindicators to manage farming systems. A set of indicators of soil biological quality are evaluated in the project: microorganisms (bacteria and fungi abundance and diversity), fauna (abundance and diversity of nematodes and earthworms), physico-chemical characteristics, soil structure assessment and degradation rate of organic matter. Based on a network of 13 plots that have been subject to an initial diagnosis in 2017, several agronomical practices to restore soil fertility are experimented to redesign the cropping system (for instance plant cover, organic matter inputs, reduction of herbicides, mineral fertilizers). System redesign was made in collaboration by winegrowers and an interdisciplinary group of experts (agronomists, biologists). Several indicators are measured on vine and soil at each vintage to assess vine health and productivity. At the end of the project (2021), a final diagnosis was carried out. Gascogn’Innov allowed to create a regional database on the quality of wine-growing soils, which permitted to evaluate the effect of practices according to soil types. Especially, decreasing the intensity of tillage and increasing the duration and diversity of grass coverage tends to increase the abundance of all the organisms studied. This project confirmed the value of soil biological quality indicators to drive the sustainability of practices, but also highlighted the key-role of expertise, in both agronomy and soil biology, to help winegrowers understand and appropriate their soil quality diagnoses.

Is wine terroir a valid concept under a changing climate?

The OIV[i] defines terroir as a concept referring to an area in which collective knowledge of the interactions between the physical and biological environment (soil, topography, climate, landscape characteristics and biodiversity features) and vitivinicultural practices develops, providing distinctive wine characteristics. Those are perceptible in the taste of wine, which drives consumer preference and, therefore, wine’s value in the marketplace. Geographical indications (GI) are recognized regulatory constructs formalizing and protecting the nexus between wine taste and the terroir generating it. Despite considering updates, GIs do not consider the nexus as a dynamic one and do not anticipate change, namely of climate. Being climate a fundamental feature of terroir, it strongly impacts wine characteristics, such as taste. According to IPCC[ii], many widespread, rapid and unprecedented changes of climate occurred, some being irreversible over hundreds to thousands of years. Climatic shifts and atmospheric-driven extreme events have been widely reported worldwide. Recent climatic trends are projected to strengthen in upcoming decades, whereas extremes are expected to increase in frequency and intensity, forcing wines away from GI definitions. Geographical shifts of viticultural suitability are projected, often moving into regions and countries different from current ones. Some authors propose adaptation in viticulture, winemaking and product innovation. We show evidence of climate changing wine characteristics in the Douro valley, home of 270-year-old Port GI. We discuss herein resist or adapt stances for when climate changes the nexus between terroir and wine characteristics. Using the MED-GOLD[iii] dashboard, a tool allowing for easy visual navigation of past and future climates, we demonstrate how policymakers can identify future moments, throughout the 21st century under different emission scenarios, when GI specifications will likely need updates (e.g., boundaries, varieties) to reduce climate-change impacts.

Climate and the evolving mix of grape varieties in Australia’s wine regions

The purpose of this study is to examine the changing mix of winegrape varieties in Australia so as to address the question: In the light of key climate indicators and predictions of further climate change, how appropriate are the grape varieties currently planted in Australia’s wine regions? To achieve this, regions are classified into zones according to each region’s climate variables, particularly average growing season temperature (GST), leaving aside within-region variations in climates. Five different climatic classifications are reported. Using projections of GSTs for the mid- and late 21st century, the extent to which each region is projected to move from its current zone classification to a warmer one is reported. Also shown is the changing proportion of each of 21 key varieties grown in a GST zone considered to be optimal for premium winegrape production. Together these indicators strengthen earlier suggestions that the mix of varieties may be currently less than ideal in many Australian wine regions, and would become even less so in coming decades if that mix was not altered in the anticipation of climate change. That is, grape varieties in many (especially the warmest) regions will have to keep changing, or wineries will have to seek fruit from higher latitudes or elevations if they wish to retain their current mix of varieties and wine styles.

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.

Modulation of berry composition by different vineyard management practices

High concentration of sugars in grapes and alcohol in wines is one of the consequences of climate change on viticulture production in several wine-growing regions. In order to investigate the possibilities of adaptation of vineyard management practices aimed to reduce the accumulation of sugar during the maturation phase without reducing the accumulation of anthocyanins in grapes, a study with severe shoot trimming, shoot thinning, cluster thinning and date of harvest was conducted on Merlot variety in Istria region (Croatia), under the Mediterranean climate. Four factors which may affect grape maturation and its composition at harvest were investigated in a two-years experiment; severe shoot trimming applied at veraison when >80% of berries changed colour (in comparison to untreated control), shoot thinning (0 and 30%), cluster thinning (0 and 30%), and the date of harvest (early and standard harvest dates). Shoot thinning had no significant impact on berry composition, despite the obtained reduction in yield per vine. Lower Brix in grapes were obtained with earlier harvest date and if no cluster thinning was applied, although at the same time a reduction in the concentration of anthocyanins in berries was observed in these treatments. On the other hand, if severe shoot trimming was applied when >80% of berries changed colour, a reduction of Brix was obtained without a negative impact on berry anthocyanins concentration. We conclude that in cases when undesirably high sugar concentrations at harvest are expected, severe shoot trimming at 80% veraison may effectively be used in order to obtain moderate sugar concentration in berries together with the adequate phenolic composition.