Macrowine 2021
IVES 9 IVES Conference Series 9 Proteomic and activity characterization of exocellular laccases from three Botrytis cinerea strains

Proteomic and activity characterization of exocellular laccases from three Botrytis cinerea strains

Abstract

Botrytis cinerea is a fungus that causes common infection in grapes and other fruits. In winemaking, its presence can be both considered desirable in the case of noble rot infection or undesirable when grey rot is developed. This fungus produces an extracellular enzyme known as laccase which is able to cause oxidation of phenolic compounds present in must and wine, causing most of the times a decrease in its quality and problems during the winemaking process [1]. Material and methods: Three B. cinerea strains (B0510, VA612 and RM344) were selected and grown in a liquid medium adapted from one previously described [2]. The enzyme was isolated by tangential ultrafiltration of the culture medium using a QuixStand system equipped with a 30 KDa filtration membrane. The purity of the isolated enzymes was checked using SDS-PAGE. The characteristics (molecular weight, % of glycosylation, specific activity, activity in function of pH of the enzyme isolated from each strain were studied with ABTS as a substrate. Results: The enzymes isolated from the three strains showed the same molecular weight, 97 KDa, in good agreement with the molecular weight previously determined for B. cinerea laccase using SDS-PAGE [2]. The percentage of glycosylation was high, being estimated in 70% on weight, also similar to that described by other authors [3]. Despite similar physical characteristics of the enzymes obtained from different strains, their activity were quite different. The enzymes isolated from B0510 and VA612 strains showed similar specific activity for ABTS oxidation, being 0.3 and 0.21 mM for their Km and their Vmax were 1.28 and 1.45 mM/min per milligram of enzyme respectively. The activity for RM344 enzyme was found much lower, with values of 0.78 mM for Km and 0.13 mM/min per milligram of enzyme for Vmax. The enzyme isolated from the B0510 strain presented its highest activity at pH 2.9 while VA612 and RM344 enzymes showed the maximum activity at pH 3.3. All these values were quite lower compared to previously measured by other authors [4-6]. Those differences in the enzyme activity may be related with differences in the active center of the enzyme and could have important consequences for the winemaking process depending on the strains of the B. cinerea strain involved in the infection of grapes.

[1]P. Ribéreau-Gayon, Y. Glories, A. Maujean, D. Dubourdieu, Handbook of Enology, Volume 2: The chemistry of Wine Stabilization and Treatments, John Wiley & Sons, Ltd, 2000. [2]D. Slomczynski, J.P. Nakas, S.W. Tanenbaum, Applied and Environmental Microbiology, 61 (1995) 907. [3]C. THURSTON, Microbiology-Sgm, 140 (1994) 19. [4]I. MARBACH, E. HAREL, A. MAYER, Phytochemistry, 23 (1984) 2713. [5]I. MARBACH, E. HAREL, A. MAYER, Phytochemistry, 22 (1983) 1535. [6]M. Dubernet, P. Ribereau-Gayon, H.R. Lerner, E. Harel, A.M. Mayer, Phytochemistry, 16 (1977) 191.

Publication date: May 17, 2024

Issue: Macrowine 2016

Type: Poster

Authors

Cédric Saucier*, Anne-Sophie Walker, Christiane Auclair, François Garcia, Francois-Xavier Sauvage, Jullien Drone, Natalia Quijada-Morin, Patrick Chemardin

*Université de Montpellier

Contact the author

Tags

IVES Conference Series | Macrowine | Macrowine 2016

Citation

Related articles…

Study of the content of amino acids and biogenic amines in sparkling red wines

The production of red sparkling wines is lower in Spain in comparison with the winemaking of white or rosé sparkling wines. In red sparkling wine processing it is essential to obtain suitable base wines that should have moderate alcohol content, high acidity, good color values, an adequate mouth-feel and a sweet tannin. Grapes for sparkling wine production have to be harvested at low maturity stages, with lower alcohol contents and higher acidities, which will that the phenolic maturity of the grapes is also low, showing green tannins. This paper analyses different treatments in order to minimize these inconveniences: cold maceration-prefermentation and delestage to elaborate the grapes with lower maturity, must nanofiltration, and the partial osmosis of the wines made from grapes with an adequate maturity degree.

Comparison of various storage conditions to preserve polyphenols in red-grape pomace

Red grape pomace, a waste from wine production, can be valorised by extracting polyphenols, high-added value compounds used in cosmetics or oenology. For use at an industrial level, using green extraction techniques, pomace need to be stored before being processed. The aim of this study is to test various storage conditions in order to maintain high level of polyphenols over 180 days, while keeping storage cost economically interesting. In a first step, different storage conditions (ambient temperature or cooled (4°C) temperature, anaerobic (saturation with N2) or aerobic conditions, and addition of sulphur dioxide (SO2)) were compared on small samples (1 kg) packed in plastic pockets. The quality of storage was assessed by following the optical density of the pomace extract at 280 nm (DO 280 expressed as mg/l eq gallic acid), which is an indication of the amount of remaining extractable polyphenols.

Ethyl esters interact with the major wine Thaumatin Like Protein VVTL1

The interactions among aromatic compounds and proteins is an important issue for the quality of foods and beverages. In wine, the loss of flavor after vinification is associated to bentonite treatment and this effect can be the result of the removal of aroma compounds which are bound wine proteins. This phenomenon was recently demonstrated for long chain fatty acids and their ethyl esters (1). Since these latter compounds are spectroscopically silent, their association with proteins is not easy to measure.

Field-grown Sauvignon Blanc berries react to increased exposure by controlling antioxidant homeostasis and displaying UV acclimation responses that are influenced by the level of ambient light

Leaf removal in the bunch zone is a common viticultural practice with several objectives, yet it has been difficult to conclusively link the physiological mechanism(s) and metabolic berry impact to this widely practiced treatment. We used a field-omics approach1 in a Sauvignon blanc high altitude model vineyard, showing that the early leaf removal in the bunch zone caused quantifiable and stable responses (over years) in the microclimate where the main perturbation was increased exposure. We provide an explanation for how leaf removal leads to the shifts in grape metabolites typically linked to this treatment and confirm anecdotal evidence and previous reports that leaf removal treatment at an early stage of berry development affects “quality-associated” metabolites (monoterpenes and norisoprenoids).

The impact of different yeasts and harvest time on the wine quality of Beihong and Beimei (<I>V. vinifera x V. amurensis</I>)

Beihong and Beimei are two wine cultivars from ‘Muscat Hamberg’ (V. vinifera L.) and wild V. amurensis Rupr., which were released in China in 2008. Here,two enology practices were reported. Firstly, the impact of different yeasts including D254, GRE, K1, D21 and BDX on dry wine quality of Beihong and Beimei was investigated. For Beihong, among wines fermented by all yeasts, residual sugar content was the lowest, total anthocyanin and resveratrol contents were the highest in the wine by D254. However, the wine by D254 had lower titrable acid than those by the other yeasts except BDX.