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

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.

The aromas found in young Bordeaux red wines made with Merlot and Cabernet Sauvignon suggest a complex mixture of aromas of fresh red fruits such as cherry or blackberry for Merlot, and strawberry or blackcurrant for Cabernet Sauvignon. The aromas of these wines are closely linked with the maturity of the grapes. The climate change that has occurred during the last decade in Bordeaux has induced changes in the ripening conditions of grape berries. It is now widely admitted that over-ripening of the berries during hot and dry summers results in the development of characteristic flavors reminiscent of cooked fruits (fig, prune). The presence of these overriding odors found in both musts and young wines affects the quality and subtlety of the wine flavor and may shorten its shelf life.

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.

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.

Comprehensive two-dimensional gas chromatography (GCxGC) has emerged as a powerful analytical technique for unraveling the volatile composition of complex matrices. This work will present three applications of GCxGC Tof-MS to the oenological field, aimed to identify novel biomarkers to be used in the quality control process of the wine industry. Comprehensive mapping of volatile compounds was conducted in a large sample of 70 sparkling wines, produced by 48 different wineries across 6 vintages and representative of the two main production areas for premium Italian sparkling wines (Franciacorta (FC) and Trentodoc (TN)), using HS-SPME followed by GCxGC-Tof-MS and multivariate analysis. Selection and identification of 196 putative biomarkers allowed clear separation of sparkling wines from FC and TN.