Flavanol glycosides in grapes and wines : the key missing molecular intermediates in condensed tannin biosynthesis ?

Since 2012, EU Commission approved compulsory labeling of wines treated with allergenic additives or processing aids “if their presence can be detected in the final product” (EU Commission Implementing Regulation No. 579/2012 of 29 June 2012). The list of potential allergens to be indicated on wine labels comprises sulphur dioxide and milk- and egg- derived fining agents, including hen egg lysozyme, which is usually added in wines as preservative. In some non-EU countries, the list includes gluten, tree nuts and fish gelatins. With the exception of lysozyme, all these fining proteins were long thought to be totally removed by subsequent winemaking processings (e.g. bentonite addition).

All techniques usually used to assay proteins was not reliable in vegetable extract due to interferences with the components included in extracts like polyphenols, tanins, pectines, aromatics compounds. Absorbance at 280nm, Kjeldhal assay, Biuret and Lowry methods, Acid Bicinchonique technique and Bradford assay give the results depending on the composition of extract, on the presence or not of detergent and on the raw material (Marchal, 1995). Another difficulty in these extracts for the quantification of proteins comes from the large amount of water included in vegetable and the low concentration of proteins. Thus in red wines, proteins are usually not taken into account due to their low concentration (typically below 10 mgL-1) and to the presence of anthocyanis and polyphenols.

Nitrogen is an important nutrient of yeast and its low content in grape must is a major cause for sluggish fermentations. To prevent problems during fermentation, a supplementation of the must with ammonium salts or more complex nitrogen mixtures is practiced in the cellar. However this correction seems to improve only partially the quality of wine [1]. In fact, yeast is using nitrogen in many of its metabolic pathways and depending of the sort of the nitrogen source (ammonium or amino acids) it produces different flavor active compounds. A limitation in amino acids can lead to a change in the metabolic pathways of yeast and consequently alter wine quality.

Tannin, anthocyanins and their reaction products play a major role in the quality of red wines. They contribute to their sensory characteristics, particularly colour and astringency. Grape tannins and anthocyanins are extracted during red wine fermentation. However, their concentration and composition change over time, due to their strong chemical reactivity1. It is also well known that yeasts influence the wine phenolic content, either through the release of metabolites involved in the formation of derived pigments1, or through polyphenol adsorption2,3.

Climate change and harvest date decisions have led to the evolution of must quality over the last decades. Increases in must sugar concentrations are among the most obvious consequences, quantitatively. Saccharomyces cerevisiae is a robust and acid tolerant organism. These properties, its sugar to ethanol conversion rate and ethanol tolerance make it the ideal production organism for wine fermentations. Unfortunately, high sugar concentrations may affect S. cerevisiae and lead to growth inhibition or yeast lysis, and cause sluggish or stuck fermentations. Even sublethal conditions cause a hyperosmotic stress response in S. cerevisiae which leads to increased formation of fermentation by-products, including acetic acid, which may exceed legal limits in some wines.