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.

A part, at least, of the fruity aroma of red wines is the consequence of perceptive interactions between various aromatic compounds, particularly ethyl esters and acetates, which may contribute to the perception of fruity aromas, specifically thanks to synergistic effects.1,2 The question of the indirect impact of non-fruity compounds on this particular aromatic expression has not yet been widely investigated. Among these compounds higher alcohols (HA) represent the main group, from a quantitative standpoint, of volatiles in many alcoholic beverages. Moreover, some bibliographic data suggested their contribution to the aromatic complexity by either increasing or masking flavors of wine, depending of their concentrations.

Polyphenols are present in a wide variety of plants and foods such as tea, cacao and grape1. An important sub-class of these compounds is the flavanols present in grapes and wines as monomers (e.g (+)-catechin or (-)-epicatechin), or polymers also called condensed tannins or proanthocyanidins. They have important antioxidant properties2 but their biosynthesis remains partly unknown. Some recent studies have focused on the role of glycosylated intermediates that are involved in the transport of the monomers and may serve as precursors in the polymerization mechanism3, 4. The global objective of this work is to identify flavanol glycosides in grapes or wines, describe their structure and determine their abundance during grape development and in wine.

Vitis vinifera is one of the most diffused grapevines over the word and it is the raw material for high quality wines production. The availability of more resistant interspecific hybrid vine varieties, developed from crosses between Vitis vinifera and other Vitis species, has generating much interest, also due to the low environmental effect of production. However, hybrid grape wine composition and varietal differences between interspecific hybrids are not well defined. Different studies revealed that wine consumption has health effects due to its high content of antioxidants, as phenolic compounds. In particular, simple phenols are appreciated not only for their physiological health benefits, including antioxidant, anti-inflammatory and cardioprotective effects, but also because they affect wines organoleptic profile and have a significant role in defining their nutritional characteristics.

The use of next-generation sequencing (NGS) has helped understand microbial genetics in oenology. Current studies mainly focus on barcoded amplicon NGS but not shotgun sequencing, which is useful for functional analyses. Since the high percentage of grapevine DNA conceals the microbial DNA in must, the majority of sequencing data is wasted in bioinformatic analyses. Here we present capture depletion of grapevine whole genome DNA.