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

Tannin and polysaccharide concentration and composition is important in defining the texture of red wines, but can vary due to factors such as cultivar, region, grape ripeness, viticultural practices and winemaking techniques. However, the concentration and composition of these macromolecules is dependent not only on grape tannin and polysaccharide concentration and composition, but also their extractability and, in the case of polysaccharides, their formation by yeast. Through studies into the influence of grape maturity, winemaking and sensory impacts of red grape polysaccharides, seed and skin tannins, recent research in our laboratory has shown that the processes involved in the extraction of these macromolecules from grapes and their retention in wine are very complex.

Laboratório de Análisis del Aroma y Enologia (LAAE). Department of Analytical Chemistry, Faculty of Sciences, Universidad de Zaragoza, 50009, Zaragoza, Spain, During alcoholic fermentation, fusel (or Strecker) aldehydes are intermediates in the amino acid catabolism to form fusel alcohols following the Ehrlich Pathway (1). One of the main enzymes involved in this pathway is Alcohol Dehydrogenase (ADH), whose activity is highly strain dependent and determines the rate of conversion of aldehydes into fusel alcohols (2). This enzyme has a Zn2+ catalytic binding site, which suggests that the must Zn2+ levels will most likely influence the rate of reduction of aldehydes into alcohols. On the other hand, SO2 is commonly used in winemaking for its antiseptic and antioxidant properties.

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).

“The Human being has an excellent ability to detect and discriminate odors but typically has great difficulty in identifying specific odorants”(1). Furthermore, “from a cognitive point of view the mechanism used to judge wines is closer to pattern recognition than descriptive analysis.” Therefore, when one wants to reveal the volatile “wine-like feature” pattern recognition techniques are required. Sensomics is one of the most recent “omics”, i.e. a holistic perspective of a complex system, which deals with the description of substances originated from microorganism metabolism that are “active” to human senses (2). Depicting the relevant volatile fraction in wines has been an ongoing task in recent decades to which several research groups have allocated important resources. The most common strategy has been the “target approach” in order to identify the “key odorants” for a given wine varietal.

Flavonols are a class of flavonoid compounds derived from plant secondary metabolism. There they play different roles like antioxidants, internal regulators and UV screenings. In red wines, flavonols have increasingly received consideration by part of scientific and winemakers according their properties began to arise known. Among these stand out wine colour stabilization and their value as bioactive compounds. In this work the complete series of the acetylated and p-coumaroylated derivatives of the 3-O-glycosides of methoxylated flavonols, namely isorhamnetin, laricitrin and syringetin, have been identified in grapes and their respective wines from Vitis vinifera cv. Tannat.