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

Del Barrio-Galán, R.a, b, Gómez-Parrini, A.a, Peña-Neira, A.b a Lallemand Inc. Chile y Compañía Limitada, Rosario Norte 407, piso 6, Las condes, Santiago, Chile b Department of Agro-Industry and Enology, Faculty of Agronomical Sciences, University of Chile, Post Office Box 1004, Santa Rosa 11315, La Pintana, Santiago, Chile It is well known that polysaccharides, mainly mannoproteins, play an important role on physical, chemical and sensory quality of wines. The ageing of white wines on lees is used in order to release higher amounts of polysaccharides by the autolytic processes in order to obtain higher-quality wines. However, this technique is too slow, because the temperature and pH conditions are not the most suitable for this process. In addition, it can also involve certain disadvantages such as a greater demand on winery resources, a longer period of wine storage, the appearance of reduction notes and some microbiological alterations.

Those previous years, pesticides are often brought to the forefront by media. Questions arose about their toxicity for growers and consumers. Even if a downward trend is underway, the use of pesticides is required to ensure steady quality and quantity of harvests. A large number of active ingredients are authorized but regarding viticulture, mainly insecticides and fungicides are applied, to control pests and diseases and to increase crop yield. Some phytosanitary products, principally fungicides, applied close to the harvest date may frequently be detected in wines.

Chemical studies aiming at assessing how a wine reacts towards oxidation usually focus on the characterization of wine constituents, such as polyphenols, or oxidation products. As an alternative, the key oxidation intermediate hydrogen peroxide H2O2 has never been quantified, although it plays a pivotal role in wine oxidation. H2O2 is obtained from molecular oxygen as the result of a first cascade of oxidation reactions involving metal ions and polyphenols. The produced H2O2 then reacts in a second cascade of oxidation to produce reactive hydroxyl radicals that can attack almost any chemical substrate in wine.

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.

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.