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

Lately several works highlighted the capacity of grape cell-wall material (CWM) to interact with proanthocyanidins (PA), indicating its potential use as fining agent for red wines.1–4 However, those studies were performed by using purified PAs and very high doses of CWM (almost ten-fold higher than those used in wine industry for other commercial fining agents). The present study focuses on the applicability of CWM from Cabernet sauvignon pomace as fining agent for red wines under real winery conditions. Grapes of cultivar Cabernet sauvignon were harvested at three different maturity levels
(unripe, mature, and overripe) and used for red winemaking. The pomace of such vinifications were used as source of CWM, and applied into red wines at two different concentrations: 0.2 g/L and 2.5 g/L.

Introduction: Malolactic fermentation (MLF) is a common process in winemaking to reduce wine acidity, maintain microbial stability and modify wine aroma. However, successful MLF is often hampered by their sluggish or stuck activity of malolactic bacteria (MLB) which may be caused by nutrient deficiency, especially when MLB are inoculated after alcoholic fermentation (Alexandre et al., 2004; Lerm et al., 2010). Identification and characterization of essential nutrients and growth factors for MLB allows for production of highly efficient nutrient supplements for MLF.

The use of bentonite in oenology rounds around the limpidity and the stability that determine consumer acceptability. As a matter of fact, the haze formation in wine reduces its commercial value and makes it unacceptable for sale. Stabilization treatments are, therefore, essential to ensure a long-time limpidity and to forecast the formation of deposits in the bottle. Bentonite that is normally used in oenology for clarifying-fining purpose, shows a natural clay-based mineral structure allowing it to swell and to jelly in water and hence in must and wine.

Nutrient availability – nitrogen, lipids, vitamins or oxygen – has a major impact on the kinetics of winemaking fermentations. Nitrogen is usually the growth-limiting nutrient and its availability determines the fermentation rate, and therefore the fermentation duration. In some cases, in particular in Champagne, grape musts have high nitrogen concentrations and are sometimes clarified with turbidity below 50 NTU. In these conditions, lipid deficiencies may occur and longer fermentations can be observed. To better understand this situation, a study was realized using a synthetic medium simulating the composition of a Champagne must : 180 g/L of sugar, 360 mg/L of assimilable nitrogen and a lipid content ranging from 1 to 8 mg/L of phytosterols (mainly β-sitosterol).

A method of suspect screening analysis to study grape metabolomics, was developed [1]. By performing ultra-high performance liquid chromatography (UHPLC) – high-resolution mass spectrometry (HRMS) analysis of the grape extract, averaging 320-450 putative grape compounds are identified which include mainly polyphenols. Identification of metabolites is performed by a new HRMS-database of putative grape and wine compounds expressly constructed (GrapeMetabolomics) which currently includes around 1,100 entries.