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

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.

Vitis vinifera L. is the most widely cultivated Vitis species which includes numerous cultivars. Owing to their superior quality of grapes, these cultivars were long considered the only suitable for the production of fine wines. However, the lack of resistance genes in V. vinifera against major grapevine pathogens, requires for its cultivation frequent spraying with large amount of fungicides. Thus, the search for alternative and more sustainable methods to control the grapevine pathogens have brought the breeders to focus their attention on other Vitis species. In fact, wild Vitis genotypes present multiple resistance traits against pathogens, such as powdery mildew, downy mildew and phylloxera.

Astringency sensation decreases slowly during the aging of red wine. Complex reactions of condensation and precipitation of wine polyphenols are involved in this phenomenon. Wine composition and conditions of aging, such as temperature and oxygen availability, strongly influence evolution of the phenol matrix. Recently, a Post-Fermentative cold Maceration (PFM) technique was tested with the aim of accelerating reactions leading to the reduction of astringency and exploiting chemical compounds not extracted from the solid parts of grapes during the previous traditional maceration phase. To this purpose, an innovative maceration system was engineered and used to perform PFM trials on marc derived from vinification of different varieties of red grapes.

The chemical mechanisms involved in oxidation/reduction potential of wine during winemaking and aging are affecting its color, aroma and taste. Chemical oxidation is one of the major causes of development of off-flavors during ageing1. Thus, the chemical changes in wine during storage should be controlled to ensure the sensory quality of the product and avoid consumer rejection that will compromise the economic value of the product. The 1-hydroxyethyl radical has been recognized as the key radical intermediate in the oxidative reactions in wine2. Based on the kinetic study of POBN-1-hydroxyethyl spin adduct formation in wines initiated via the Fenton reaction, a novel tool was recently developed in our laboratory to quantify the resistance of wines against oxidation3.

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.