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…

Measurement of redox potential as a new analytical winegrowing tool

Excell laboratory has initiated the development of an analytical method based on electrochemistry to evaluate the ability of wines to undergo or resist to oxidative phenomena. Electrochemistry is a powerful tool to probe reactions involving electron transfers and offers possibility of real-time measurements. In that context, the laboratory has implemented electrochemical analysis to assess oxidation state of different wine matrices but also in order to evaluate oxidative or reduced character of leaf and soil. Initially, our laboratory focused on dosage of compounds involved in responses of plant stresses and we were also interested in microbiological activity of soils. These analyses were compared with the measurement of redox potential (Eh) and pH which are two fundamental variables involved in the modulation of plant metabolism. Indeed, the variation of redox states of the plant reflects its biological activity but also its capacity to absorb nutriments. The Eh-pH conditions mainly determine metabolic processes involved in soil and leaf and our goal is to determine if this combined analytical approach will be sufficiently precise to detect biological evolutions (plant health, parasitic attack…).

Influence of grapevine rootstock/scion combination on rhizosphere and root endophytic microbiomes

Soil is a reservoir of microorganisms playing important roles in biogeochemical cycles and interacting with plants whether in the rhizosphere or in the root endosphere. The composition of the microbial communities thus impacts the plant health. Rhizodeposits (such as sugar, organic and amino acids, secondary metabolites, dead root cells …) are released by the roots and influence the communities of rhizospheric microorganisms, acting as signaling compounds or carbon sources for microbes. The composition of root exudates varies depending on several factors including genotypes. As most of the cultivated grapevines worldwide are grafted plants, the aim of this study was to explore the influence of rootstock and scion genotypes on the microbial communities of the rhizosphere and the root endosphere. The work was conducted in the GreffAdapt plot (55 rootstocks x 5 scions), in which the 275 combinations have been planted into 3 blocks designed according to the soil resistivity. Samples of roots and rhizosphere of 10 scion x rootstock combinations were first collected in May among the blocks 2 and 3. The quantities of bacteria, fungi and archaea have been assessed in the rhizosphere by quantitative PCR, and by cultivable methods for bacteria and fungi. The communities of bacteria, fungi and arbuscular mycorrhizal fungi (AMF) was analyzed by Illumina sequencing of 16S rRNA gene, ITS and 28S rRNA gene, respectively. The level of mycorrhization was also evaluated using black ink coloration of newly formed roots harvested in October. The level of bacteria, fungi and archaea was dependent on rootstock and scion genotypes. A block effect was observed, suggesting that the soil characteristics strongly influenced the microorganisms from the rhizosphere and root endosphere. High-throughput sequencing of the different target genes showed different communities of bacteria, fungi and AMF associated with the scion x rootstock combinations. Finally, all the combinations were naturally mycorrhized. The root mycorrhization intensity was influenced by the rootstock genotype, but not by the scion one. Altogether, these results suggest that both rootstock and scion genotypes influence the rhizosphere and root endophytic microbiomes. It would be interesting to analyze the biochemical composition of the rhizodeposition of these genotypes for a better understanding of the processes involved in the modulation of these microbiomes. Moreover, crossing our data with the plant agronomic characteristics could provide insights into their roles on plant fitness.

Influence of agronomic practices in soil water content in mid-mountain vineyards

In the context of LIFE project MIDMACC (LIFE18 CCA/ES/001099), several pilots have been installed in vineyards in mid mountain areas of Catalonia (NE Spain) to test well stablished agronomic practices to increase the adaptation of Mediterranean mid mountain to climate change. Soil water content (SWC) at three different depths (15, 30 and 45cm) was measured in continuum from August 2020. One pilot (WC) included a well-established green cover (GC), a new GC (NC) and a conventional soil management (CM, tilling+herbicides). NC presented an intermediate state between WC and CM, responding similarly to CM in autumn but quickly reaching similar SWC to WC, then following the same evolution till next spring, with CM presenting lower values along autumn and winter. Then vegetation activation decreased SWC in all plots, (much slower in CM, lacking GC). Sensibility to spring rains is again intermediate for NC, which joins SWC evolution of CM by the end of spring till next autumn. It is expected that NC will resemble WC more and more as its GC develops. In the pilot combining vine training (VSP vs Gobelet) and hillside management (slope vs terrace), no clear pattern could be related with these conditions. However, both terraces seem to be more sensitive to spring rains. A third pilot included new vineyards (7 and 1 year old). In the new vineyard (N), higher canopy development, a spontaneous green cover and row straw resulted in a slower SWC dynamic, not so sensitive to rains but conserving more soil water in spring and most of summer, even with presumably a higher water extraction by vines. In the newest vineyard (VN) the deepest sensor is still sensitive to rain events all over the year and SWC is always highest at this depth, revealing small water capture by vines.

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.

Permanent cover cropping with reduced tillage increased resiliency of wine grape vineyards to climate change

Majority of California’s vineyards rely on supplemental irrigation to overcome abiotic stressors. In the context of climate change, increases in growing season temperatures and crop evapotranspiration pose a risk to adaptation of viticulture to climate change. Vineyard cover crops may mitigate soil erosion and preserve water resources; but there is a lack of information on how they contribute to vineyard resiliency under tillage systems. The aim of this study was to identify the optimum combination of cover crop sand tillage without adversely affecting productivity while preserving plant water status. Two experiments in two contrasting climatic regions were conducted with two cover crops, including a permanent short stature grass (P. bulbosa hybrid), barley (Hordeum spp), and resident vegetation under till vs. no-till systems in a Ruby Cabernet (V. vinifera spp.) (Fresno) and a Cabernet Sauvingon (Napa) vineyard. Results indicated that permanent grass under no-till preserved plant available water until E-L stage 17. Consequently, net carbon assimilation of the permanent grass under no-till system was enhanced compared to those with barley and resident vegetation. On the other hand, the barley under no-till system reduced grapevine net carbon assimilation during berry ripening that led to lower content of nonstructural carbohydrates in shoots at dormancy. Components of yield and berry composition including flavonoid profile at either site were not adversely affected by factors studied. Switching to a permanent cover crop under a no-till system also provided a 9% and 3% benefit in cultural practices costs in Fresno and Napa, respectively. The results of this work provides fundamental information to growers in preserving resiliency of vineyard systems in hot and warm climate regions under context of climate change.