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…

A combination of biotechnology tools and coopers elements for an alternative the addition of SO2 at the end of the malolactic fermentation in red wines or at the “mutage” for the “liquoreux” wines

In red wines the post-MLF SO2 addition is an essential event. It is also the case for the “mutage” during the elaboration of the “liquoreux”. At these moments SO2 plays an antimicrobial action and an antioxidant effect. But at current pH of wines, ensuring a powerful molecular SO2 has become very difficult. Recent work on Brettanomyces strains have also shown that some strains are resistant up to 1.2 mg / L of molecular SO2. It’s also the case of the some Saccharomuces or Zygosaccharomyces strains suitable to re-ferment “liquoreux” wines after the “mutage”.

Oxygen consumption by diferent oenological tanins in a model wine solution

INTRODUCTION: Oenological tannins are widely used in winemaking to improve some characteristics of wines [1] being the antioxidant properties probably one of the main reasons [2]. However, commercial tannins have different botanical sources and chemical composition [3] which probably determines different antioxidant potential. There are some few references about the antioxidant properties of commercial tannins [4] but none of them have really measured the direct oxygen consumption by them. The aim of this work was to measure the kinetics of oxygen consumption by different commercial tannins in order to determine their real capacities to protect wine against oxygen. MATERIAL AND METHODS: 4 different commercial tannins were used: T1: condensed tannin from grape seeds, T2: gallotannin from chinese gallnuts, T3: ellagitannin from oak and T4: tannin from quebracho containing condensed tannins and ellagitannins.

Fractionation of copper and iron in wine: Assessment of potential macromolecule and sulfur binding agents

Copper and iron are known to substantially impact wine stability through oxidative, reductive or colloidal phenomena. However, the binding of metal ions to different wine components under wine conditions, and the impact of this binding on the ability of the metal ions to induce spoilage processes, is not well understood. This study surveyed a range of red and white wines for an understanding of the variability of broad metal categories within the wines. The techniques utilized included an electrochemical constant current stripping potentiometry technique (ccSP), and solid phase extraction (SPE) fractionation of wine with subsequent analysis of the metal content of each fraction by inductively coupled plasma – optical emission spectroscopy (ICP-OES).

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.

Sensory impacts of the obturator used for the Chasselas: study over the time

Many parameters affect the organoleptic characteristics of wine: internal parameters like the chemical composition or polyphenol content and external as for example storage conditions or the type of obturator. The aim of this study was to characterize sensorally the impacts of several type of obturator on a white wine: Chasselas. To determine the organoleptic characteristics of this wine, a quantitative descriptive analysis could be used. But rapid sensory methods were preferred in this project. Indeed these methods are an appropriate alternative to conventional descriptive methods for quickly assessing sensory product discrimination.