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

The present study was performed in a traditional winery located in the viticultural area of Brunello di Montalcino, Siena, Italy, in the vintage 2015. Actually, in this winery Sangiovese grape musts are fermented in large oak barrels by a single strain of Saccharomyces cerevisiae previously isolated in the same winery. Pumping over operations are carried out once or twice a day until the end of alcoholic fermentations. The aim of this work was to investigate on the oenological properties of Sangiovese wine produced with the traditional winemaking process adopted by the winery under study obtained from the fermentation of whole berries compared to that from crushed grape must. In particular, two lots of 65q of Sangiovese grapes from the same 3ha vineyard were vinified in 150hL oak barrels.

Oak barrels form an integral part of wine production, especially that of high quality wines. However, due to its porosity, wood presents an ecological niche for microbial proliferation and is highly susceptible to microbial spoilage which could cause considerable economic losses. Brettanomyces bruxellensis, the most commonly encountered microorganism responsible for spoilage during barrel ageing, can remain in barrels after barrel sanitation to contaminate new batches of wine after refilling. Therefore, effective sanitation treatments are of utmost importance to prevent recurring wine spoilage.

Ellagitannins (ETs) have been reported to be the main phenolic compounds found in oak wood. These compounds, belonging to the hydrolysable tannin class of polyphenols, are esters of hexahydroxydiphenic acid (HHDP) and a polyol, usually glucose or quinic acid. They own their name to their capacity to be hydrolysed and liberate ellagic acid and they have an impact on astringency and bitterness sensation, which is strongly dependant on their structure. The toasting phase is particularly crucial in barrels fabrication and influences wood composition.

Copper and iron are known to substantially impact wine stability through oxidative, reductive or colloidal phenomena. However, the binding of metal ions to different wine components under wine conditions, and the impact of this binding on the ability of the metal ions to induce spoilage processes, is not well understood. This study surveyed a range of red and white wines for an understanding of the variability of broad metal categories within the wines. The techniques utilized included an electrochemical constant current stripping potentiometry technique (ccSP), and solid phase extraction (SPE) fractionation of wine with subsequent analysis of the metal content of each fraction by inductively coupled plasma – optical emission spectroscopy (ICP-OES).

Despite the extensive research performed during the last decades, the multifactorial mechanism responsible for the white wine protein haze formation is not fully characterized. Herein, a new model is proposed, which is based on the experimental identification of sulfur dioxide as a major modulating factor inducing wine protein haze upon heating. As opposed to other reducing agents, such as 2-mercaptoethanol, dithiothreitol and tris(2-carboxyethyl)phosphine hydrochloride (TCEP), the addition of SO2 to must/wine upon heating cleaves intraprotein disulfide bonds, hinders thiol-disulfide exchange during protein interactions and can lead to the formation of novel inter/intraprotein disulfide bonds. Those are eventually responsible for wine protein aggregation which follows a nucleation-growth kinetic model as shown by dynamic light scattering [1].