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…

Deconstructing the soil component of terroir: from controversy to consensus

Wine terroir describes the collectively recognized relation between a geographical area and the distinctive organoleptic characteristics of the wines produced in it. The overriding objective in terroir studies is therefore to provide scientific proof relating the properties of terroir components to wine quality and typicity. In scientific circles, the role of climate (macro-, meso- and micro-) on grape and wine characteristics is well documented and accepted as the most critical. Moreover, there has been increasing interest in recent years about new elements with possible importance in shaping wine terroir like berry/leaf/soil microbiology or even aromatic plants in proximity to the vineyard conferring flavors to the grapes. However, the actual effect of these factors is also dependent on complex interactions with plant material (variety/clone, rootstock, vine age) and with human factors. The contribution of soil, although a fundamental component of terroir and extremely popular among wine enthusiasts, remains a much-debated issue among researchers. The role of geology is probably the one mostly associated by consumers with the notion of terroir with different parent rocks considered to give birth to different wine styles. However, the relationship between wine properties and the underlying parent material raises a lot of controversy especially regarding the actual existence of rock-derived flavors in the wine (e.g. minerality). As far as the actual soil properties are concerned, the effect of soil physical properties is generally regarded as the most significant (e.g sandy soils being associated with lighter wines while those on clay with colored and tannic ones) mostly through control of water availability which ultimately modifies berry ripening conditions either directly by triggering biosynthetic pathways, or indirectly by altering vigor and yield components. The role of soil chemistry seems to be weakly associated to wine sensory characteristic, although N, K, S and Ca, but also soil pH, are often considered important in the overall soil effect. Recently, in the light of evidence provided by precision agriculture studies reporting a high variability of vineyard soils, the spatial scale should also be taken into consideration in the evaluation of the soil effects on wines. While it is accepted that soil effects become more significant than climate on a local level, it is not clear whether these micro-variations of vineyard soils are determining in the terroir effect. Moreover, as terroir is not a set of only natural factors, the magnitude of the contribution of human-related factors (irrigation, fertilization, soil management) to the soil effect still remains ambiguous. Lastly, a major shortcoming of the majority of works about soil effects on wine characteristics is the absence of connection with actual vine physiological processes since all soil effects on grape and wine chemistry and sensorial properties are ultimately mediated through vine responses. This article attempts to breakdown the main soil attributes involved in the terroir effect to suggest an improved understanding about soil’s true contribution to wine sensory characteristics. It is proposed that soil parameters per se are not as significant determining factors in the terroir effect but rather their mutual interactions as well as with other natural and human factors included in the terroir concept. Consequently, similarly to bioclimatic indices, composite soil indices (i.e. soil depth, water holding capacity, fertility, temperature etc), incorporating multiple soil parameters, might provide a more accurate and quantifiable means to assess the relative weight of the soil component in the terroir effect.

VINIoT – Precision viticulture service

The project VINIoT pursues the creation of a new technological vineyard monitoring service, which will allow companies in the wine sector in the SUDOE space to monitor plantations in real time and remotely at various levels of precision. The system is based on spectral images and an IoT architecture that allows assessing parameters of interest viticulture and the collection of data at a precise scale (level of grape, plant, plot or vineyard) will be designed. In France, three subjects were specifically developed: evaluation of maturity, of water stress, and detection of flavescence dorée. For the evaluation of maturity, it has been decided first to work at the berry scale in the laboratory, then at the bunch scale and finally in the vineyard. The acquisition of the spectral hyperstal image as well as the reference analyzes to measure the maturity, were carried out in the laboratory after harvesting the berries in a maturity monitoring context. This work focuses on a case study to predict sugar content of three different grape varieties: Syrah, Fer Servadou and Mauzac. A robust method called Roboost-PLSR, developed in the framework of this work (Courand et al., 2022), to improve prediction model performance was applied on spectra after the acquirement of hyperspectral images. Regarding the evaluation of water stress, to work with a significant variability in terms of water status, it has been worked first with potted plants under 2 different water regimes. The facilities have allowed the supervision of irrigation and micro-climatic conditions. The regression models on agronomic variables (stomatal conductance, water potential, …) are studied. To detect flavescence dorée, the experimental plan has consisted of work at leaf scale in the laboratory first, and then in the field. To detect the disease from hyper-spectral imaging, a combination of multivariate curve resolution-alternating least squares (MCR-ALS) and factorial discriminant analysis (FDA) was proposed. This strategy proved the potential towards the discrimination of healthy and infected leaves by flavescence dorée based on the use of hyperspectral images (Mas Garcia et al., 2021).

Elevational range shifts of mountain vineyards: Recent dynamics in response to a warming climate

Increasing temperatures worldwide are expected to cause a change in spatial distribution of plant species along elevational gradients and there are already observable shifts to higher elevations as a consequence of climate change for many species. Not only naturally growing plants, but also agricultural cultivations are subject to the effects of climate change, as the type of cultivation and the economic viability depends largely on the prevailing climatic conditions. A shift to higher elevations therefore represents a viable adaptation strategy to climate change, as higher elevations are characterized by lower temperatures. This is especially important in the case of viticulture because a certain wine-style can only be achieved under very specific climatic conditions. Although there are several studies investigating climatic suitability within winegrowing regions or longitudinal shifts of winegrowing areas, little is known about how fast vineyards move to higher elevations, which may represent a viable strategy for winegrowers to maintain growing conditions and thus wine-style, despite the effects of climate change. We therefore investigated the change in the spatial distribution of vineyards along an elevational gradient over the past 20 years in the mountainous wine-growing region of Alto Adige (Italy). A dataset containing information about location and planting year of more than 26000 vineyard parcels and 30 varieties was used to perform this analysis. Preliminary results suggest that there has been a shift to higher elevations for vineyards in general (from formerly 700m to currently 850 m a.s.l., with extreme sites reaching 1200 m a.s.l.), but also that this development has not been uniform across different varieties and products (i.e. vitis vinifera vs hybrid varieties and still vssparkling wines). This is important for climate change adaptation as well as for rural development. Mountain areas, especially at mid to high elevations, are often characterized by severe land abandonment which can be avoided to some degree if economically viable and sustainable land management strategies are available.

Effect of vigour and number of clusters on eonological parameters and metabolic profile of Cabernet Sauvignon red wines

Vegetative growth and yield are reported to affect grape and wine quality. They can be controlled through different techniques linked to vine management. The objective of this research was to determine the effect of vine vigour and number of clusters per vine on physicochemical composition and phenolic profile of red wines. The experiment was carried out during two vegetative cycles, with cv. Cabernet Sauvignon grafted onto Paulsen 1103. Three vine vigour were defined, according to shoot weight at previous harvests, being low, medium and high. Five treatments of number of clusters were used for each vigour, with 15, 22, 29, 36, and 45 clusters per vine. Grapes from all treatments were harvested in the same day from Brix and total acidity criteria. Thirty days after bottling, classical analyzes and phenolic compounds were performed. As results, different responses were obtained from each vintage. In 2020, a dry season from veraison to harvest, grapes and wines obtained from low vigour treatment and 45 clusters per vine was the highest in sugar and alcohol content respectively, while grapes and wines from high vigour and 15 clusters presented the lowest sugar and alcohol content. Total anthocyanins were higher in treatment with low vigour and 15 clusters, while the lowest amounts were found in low vigour with 45 clusters, as well as medium and high vigour with 36 clusters per vine. Total tannins were higher in high vigour with 22 clusters and medium vigour with 29 clusters, while were lower in low vigour with 36 clusters. In 2021, a wet season at harvest, responses were different, and great variations were observed between treatments. As conclusions, yield and vine vigour had strong influence on grape and wine quality, promoting different enological potentials on which can be indicated/used for aging strategies of red and even rosé wines.

Influence of a spontaneous cover crop on the vineyard and soil erosion under Mediterranean climate

Sixty five % of the agricultural area of the Basque Country located in the DO Ca Rioja corresponds to vineyards. More than 40% of it has an average slope greater than 10%, which makes it sensitive to erosive processes. Furthermore, it is foreseeable that extreme weather events (storms, hail, extreme heat and cold, etc.) will be favored due to climate change. Cover cropping can mitigate this risk, and therefore the objective of this work is to evaluate the impact that a vegetable cover has on the agronomic behavior of the vineyard, the quality of the grape and soil erosion. For this, a trial has been carried out with a Graciano variety vineyard with a slope between 10% -20% during the years 2020 and 2021. Conventional tillage management in the area has been compared (4-6 passes per year of tillage machinery) versus spontaneous vegetation cover management in the vineyard. This implies not tilling and allowing the grass of the land to colonize the range between the lines of vines, controlling their height through 1-3 mowing passes per year, always trying to affect the surface of the land as little as possible. The vegetative growth, yield and quality of the grape and wine was measured. Furthermore, erosion has been measured using Gerlasch boxes. The yield was lower in the second year of the trial in the cover crop treatment, but erosion was significantly reduced.