Directed Evolution of Oenococcus oeni: optimising yeast-bacteria interactions for improved malolactic fermentation

Glutathione is a tripeptide (γ-Glu-Cys-Gly), which can occur in grapes, in must and in wine prevalently in the reduced form as well as in the oxidized form as glutathione disulfide. The importance of the reduced form of glutathione lies in its antioxidant activity. In must, it limits browning by reducing o-quinones produced by polyphenol oxidase activity on hydroxycinnamic acids; in wine, it exerts a protective effect on various aromatic compounds. Glutathione concentration in wine is lower than in grape juice and variable as it depends on several factors, ranging from the native content of grapes to winemaking technique.

Grape by-products (including skins, seeds, stems and vine shoots) are rich in health promoting polyphenols. Their extraction from winery waste and their following purification are of special interest to produce extracts with high added value compounds. Meanwhile, the growing concern over environmental problems associated with economic constraints, require the development of environmentally sustainable extraction technologies. The extraction using semi-continuous subcritical water, as a natural solvent at high temperature and high pressure a technology is promising “green” technology that is environmentally friendly, energy efficient and improve the extraction process in plant tissues.

The aim of this work was to reduce the alcohol content of Tempranillo wine. Tempranillo wines were produced by grapes harvested at different ripening dates (August 11 which was 21 oBrix and September 28 with 25 oBrix). At the second date, the Tempranillo wines were elaborated as follows: grapes were destemmed, crushed and collected into 50 L stainless-steel vats. Before preferementative maceration in cold, 50 % (M1) and 70 % (M2) of the must have been replaced by the same percentage of must from the first harvest. In addition, a control wine (C) was performed with only grapes from the second harvest.

The use of glutathione (GSH) in winemaking has been legitimated recently, according to OIV resolutions OENO 445-2015 and OENO 446-2015 a maximum dose of 20 mg/L is now allowed to use in must and wine. Several studies have proven the benefits of GSH, predominantly in Sauvignon blanc. Thus, oxidative coloration of must and wine is limited, aroma compounds such as volatile thiols are preserved, and the development of ageing flavors such as sotolon and 2-aminoacetophenone is impeded. The protective effect may be explained by the high affinity of GSH to bind o-quinones which are formed during phenolic oxidation and which are known to initiate browning and other oxidative changes. Some researchers have proposed the hydroxycinnamic acid to GSH ratio (HGR) as an indicator of oxidation susceptibility of must and could show that lower ratios yielded lighter musts.

Must oxidation is a complex process involving multiple enzymatic transformations, including the oxidation of phenolics containing an ortho-diphenol function. The latter process has a primary influence on wine aroma characteristics and stability, due to the central role of ortho-diphenols in the non-enzymatic oxidative reactions taking place during winemaking and in finished wine. Although oxidation of must is traditionally avoided, in recent years its contribution to wine quality has been revisited, and in some cases improvements to wine aroma have been observed with the application of controlled must oxidation. Nowadays there is a great interest in the wine industry towards the identification of specific markers or patterns to characterize and classify the response of grape must to oxidation.