IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Exogeneous C-S lyase enzyme, a potential tool for thiol enhancement in wine or beer?

Exogeneous C-S lyase enzyme, a potential tool for thiol enhancement in wine or beer?

Abstract

Varietal thiols are considered for years as key aroma compounds in many wines. Their main origin is the cleavage during alcoholic fermentation of S-conjugate precursors present in grapes and musts, even if the levels of precursors already identified struggle to completely explain the levels of thiols found in wine.
In this bioconversion process, yeast is the key since the cleavage of thiol precursors has been demonstrated to be due to the internal C-S lyase activity of the yeast strains. Whatever the intrinsic capacity of the yeast and the regulation mechanisms identified for the transportation of thiol precursor into the cell, the conversion yield remains very low, estimated in most cases at 1%. In this context, the use of exogenous C-S lyases could be an alternative option to reveal a larger part of the thiol aroma.
Our study focused on the characterization of a recombinant C-S lyase model obtained by from Lactobaccilus delbrueckii subsp. bulgaricus, expressed in E. coli [1] to investigate the possibility of using such enzymes in different matrices such as wine, beer or directly hops, known to be rich in thiol precursors.
A spectrophotometric method was developed for the quantification of the C-S lyase activity, using commercially available S-4-nitrophenyl-L-cysteine (Cys(4NP)). This method was then used to study the specificity of substrate and potential competitor recognition, still using Cys(4NP) but also G-4NP, Cys(4NP)-Gly and γGlu-Cys(4NP) conjugates, which were specifically synthesized in our lab, based on our previous results [2].
The C-S lyase we used was able to cleave efficiently Cys(4NP), but not glutathione and dipeptide 4NP-S-conjugates. As expected, pH emerged as a key parameter: no reaction at wine pH (2.8-3.5), low activity at beer pH (4.5-5.6) and > 80% of activity at pH above 6. Assays with N-Ac-Cys(4NP) confirmed that a free amine group on the substrate was compulsory for recognition by the enzyme and subsequent cleavage of the substrate. Free cysteine has also been demonstrated to compete with Cys(4NP) resulting in a dramatic decrease in conversion efficiency.
These first results documented the possibility of using such enzyme in the different matrices, highlighting the constraints for the subsequent identification of C-S lyase more suitable to wine or beer productions

References

[1] Allegrini, A.; Astegno, A.; La Verde, V.; Dominici, P. Characterization of C-S lyase from Lactobacillus delbrueckii subsp. bulgaricus ATCC BAA-365 and its potential role in food flavor applications. J. Biochem. 2017, 61, 349−360.
[2] Bonnaffoux, H., Roland, A., Rémond, E., Delpech, S., Schneider, R., & Cavelier, F. (2017). First identification and quantification of S-3-(hexan-1-ol)-γ-glutamyl-cysteine in grape must as a potential thiol precursor, using UPLC-MS/MS analysis and stable isotope dilution assay. Food Chemistry, 237, 877–886.

DOI:

Publication date: June 27, 2022

Issue: IVAS 2022

Type: Poster

Authors

Clerat Luigi1, Vives Eric1, Cavelier Florine2, Remond Emmanuelle2 and Schneider Rémi3

1PhyMedExp – Physiologie & médecine expérimentale du Cœur et des Muscles [U 1046]
2Institut des biomolécules Max Mousseron (IBMM) – UMR-5247 – CNRS
3Oenobrands Montpellier FR

Contact the author

Keywords

C-S lyase, varietal thiol precursors, wine aroma, S-conjugates

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

Projected changes in vine phenology of two varieties with different thermal requirements cultivated in La Mancha DO (Spain) under climate change scenarios

The aim of this work was to analyze the phenology variability of Tempranillo and Chardonnay cultivars, related to the climatic characteristics in La Mancha Designation of Origin, and their potential changes under climate change scenarios. Phenological dates referred to budbreak, flowering, veraison and harvest were analyzed for the period 2000-2019. The weather conditions at daily time scale, recorded during the same period, were also evaluated. The thermal requirements to reach each of these phenological stages were calculated and expressed as the GDD accumulated from DOY=60. Changes in phenology were projected by 2050 and 2070 taking into account those values and the projected temperatures and precipitation, simulated under two Representative Concentration Pathway (RCP) scenarios –RCP4.5 and RCP8.5– using an ensemble of models. The average phenological dates during the period under study were, April 16th ± 6.6 days and April 5th ± 6.0 days for budbreak, May 31st ± 6.0 days and May 27th ± 5.3 days for flowering, July 26th ± 5.6 days and July 25th ± 5.8 days for veraison, and Ago 23rd ± 10.8 days and Ago 17th ± 9.0 days for harvest, respectively, for Tempranillo and Chardonnay. The projected changes in temperature imply an average change in the maximum growing season (April-August) temperatures of 1.2 and 1.9°C by 2050, and 1.6 and 2.6°C by 2070, under the RCP4.5 and RCP8.5 scenarios, respectively. A reduction in precipitation is predicted, which vary between 15% for 2050 under RCP4.5 scenario and up to 30% by 2070 under RCP8.5. The advance of the phenological dates for 2050, could be of 6, 7, 7, and 8 days for Tempranillo and 4, 6, 6 and 9 days for Chardonnay, respectively for budbreak, flowering, veraison and harvest under the RCP4.5 scenario. Under the RCP8.5 emission scenario, the advance could be up to 30% higher.

Climate ethnography and wine environmental futures

Globalisation and climate change have radically transformed world wine production upsetting the established order of wine ecologies. Ecological risks and the future of traditional agricultural systems are widely debated in anthropology, but very little is understood of the particular challenges posed by climate change to viticulture which is seen by many as the canary in the coalmine of global agriculture. Moreover, wine as a globalised embedded commodity provides a particularly telling example for the study of climate change having already attracted early scientific attention. Studies of climate change in viticulture have focused primarily on the production of systematic models of adaptation and vulnerability, while the human and cultural factors, which are key to adaptation and sustainable futures, are largely missing. Climate experts have been unanimous in recognising the urgent need for a better understanding of the complex dynamics that shape how climate change is experienced and responded to by human systems. Yet this call has not yet been addressed. Climate ethnography, coined by the anthropologist Susan Crate (2011), aims to bridge this growing disjuncture between climate science and everyday life through the exploration of the social meaning of climate change. It seeks to investigate the confrontation of its social salience in different locations and under different environmental guises (Goodman 2018: 340). By understanding how wine producers make sense of the world (and the environment) and act in it, it proposes to focus on the co-production of interdisciplinary knowledge by identifying and foreshadowing problems (Goodman 2018: 342; Goodman & Marshall 2018). It seeks to offer an original, transformative and contrasted perspective to climate change scenarios by investigating human agency -individual or collective- in all its social, political and cultural diversity. An anthropological approach founded on detailed ethnographies of wine production is ideally placed to address economic, social and cultural disruptions caused by the emergence of these new environmental challenges. Indeed, the community of experts in environmental change have recently called for research that will encompass the human dimension and for more broad-based, integrated through interdisciplinarity, useful knowledge (Castree & al 2014). My paper seeks to engage with climate ethnography and discuss what it brings to the study of wine environmental futures while exploring the limitations of the anthropological environmental approach.

Impact of climate change on the viticultural climate of the Protected Designation of Origin “Jumilla” (SE Spain)

Protected Designation of Origin “Jumilla” (PDO Jumilla) is located in the Spanish provinces of Albacete and Murcia, in the South-eastern part of the Iberian Peninsula, where most of the models predict a severe impact of climate change in next decades. PDO Jumilla covers an area of 247,054 hectares, of which more than 22,000 hectares

Long-term drought resilience of traditional red grapevine varieties from a semi-arid region

In recent decades, the scarcity of water resources in agriculture in certain areas has been aggravated by climate change, which has caused an increase in temperatures, changes in rainfall patterns, as well as an increase in the frequency of extreme phenomena such as droughts and heat waves. Although the vine is considered a drought-tolerant specie, it has to satisfy important water requirements to complete its cycle, which coincides with the hottest and driest months. Achieving sustainable viticulture in this scenario requires high levels of efficiency in the use of water, a scarce resource whose use is expected to be severely restricted in the near future. In this regard, the use of drought-tolerant varieties that are able to maintain grape yield and quality could be an effective strategy to face this change. During three consecutive seasons (2018-2020) the behavior in rainfed regime of 13 traditional red grapevine varieties of the Spain central region was studied. These varieties were cultivated in a collection at Centro de Investigación de la Vid y el Vino de Castilla-La Mancha (IVICAM-IRIAF) located in Tomelloso (Castilla-La Mancha, Spain). Yield components (yield, mean bunch and berry weight, pruning weight), physicochemical parameters of the musts (brix degree, total acidity, pH) and some physiological parameters related with water stress during ripening period (δ13C, δ18O) were analysed. The application of different statistical techniques to the results showed the existence of significant differences between varieties in their response to stressful conditions. A few varieties highlighted for their high ability to adapt to drought, being able to maintain high yields due to their efficiency in the use of water. In addition, it was possible quantify to what extent climate can be a determinant in the δ18O of musts under severe water stress conditions.

Combining effect of leaf removal and natural shading on grape ripening under two irrigation strategies in Manto negro (Vitis vinifera L.)

The increasingly frequent heat waves during grape ripening pose challenges for high quality wine grape production. Defoliation is a common practice that can improve the control of diseases in bunches, but also it increases the exposure to sunlight. Grapes exposed to solar radiation reach temperatures over the optimum for berry development and maturation. This makes the development of irrigation and canopy management techniques of great importance to maximize yield and grape quality. A field experiment was carried out during 2021 using Manto negro wine grapes to study the effect of applied irrigation and different light exposure levels on grape quality. Two irrigation treatments were imposed based on the frequency and amount of water doses in a four-block experimental vineyard at Bodega Ribas (Mallorca). Three light exposure treatments were randomly applied in each irrigation plot. The light treatments included exposed clusters from pea size, non-exposed clusters, and shaded clusters after softening. Leaf area index and canopy porosity was estimated every 2 weeks. Midday leaf water potential was measured weekly. Additionally, apparent electrical conductivity was measured between rows to estimate the soil water content variability. Light and temperature sensors were installed at the bunch level to quantify the differences in bunch temperature and light intensity among treatments. The effect of irrigation and cluster light exposure on berry weight, TSS, TA, malic acid, tartaric acid, K+, and pH were analysed at 5 moments along grape ripening. During different heat waves, the natural shading technique decreased the maximum bunch temperature around 10 °C respect to the exposed bunches in both irrigation strategies. The combination of defoliation and shading techniques after softening decreased TSS at harvest and affected most of the quality parameters during the last stages of ripening, showing an interesting technique to delay ripening in warm viticulture areas.