Background and Aims: Yeast autolysis is understood to be primarily responsible for giving traditional method sparkling wines complex and developed aromas. The contribution from ageing the wine itself, however, is less well-established. This study aimed to determine the contribution of autolysis products and compounds associated with wine oxidation and ageing in Vitis vinifera L. Chardonnay and Pinot Noir wines over 24 months. Methods and Results: Chardonnay and Pinot Noir base wines were tiraged, or aged with and without primary lees. Volatile composition analyses (HS-SPME/GC/MS and GC/MS/MS) were conducted at 6, 12, and 24 months post-bottling and sensory appraisals at 12 and 24 months. The duration of ageing significantly influenced compositional changes in fermentation-derived and oxidative-flavour-associated compounds. Ageing base wines off or on lees produced similar maturation-associated aroma profiles to sparkling wines irrespective of cultivar. Conclusions: The contribution of autolysis products did not feature as strongly as anticipated over 24 months, indicating that compounds associated with wine ageing primarily influenced the aroma of mature sparkling wines. This finding suggests winemakers ageing their wines on lees for 24 months or less should place more emphasis on base wine composition to manipulate the aroma profiles of sparkling wines. Significance of the Study: First comparative chemical compositional study of base wines concurrently with sparkling wines.

Sparkling wine made by the traditional method (Méthode Traditionelle) develops a distinct and desirable flavour and aroma profile attributed to proteolytic processes during prolonged ageing on lees. Microwave, ultrasound and addition of β-glucanase enzymes were applied to accelerate the disruption of Saccharomyces cerevisiae, and added to the tirage solution for secondary fermentation in traditional sparkling winemaking. Scanning electron microscopy and flow cytometry analyses were used to observe and describe yeast whole-cell anatomy, and cell integrity and structure via propidium iodide (PI) permeability after 6-, 12- and 18-months post-tirage. Treatments applied produced features on lees that were distinct from that of the untreated control yeast. Whilst control yeast displayed budding cells (growth features) with smooth, cavitated and flat external cell appearances; microwave treated yeast cells exhibited modifications like ‘doughnut’ shapes immediately after treatment (time 0). Similar ‘doughnut’-shaped and ‘pitted/porous’ cell features were observed on progressively older lees from the control. Flow cytometry was used to discriminate yeast populations; features consistent with cell disruption were observed in the microwave, ultrasound and enzyme treatments, as evidenced by up to 4-fold increase in PI signal in the microwave treatment. Forward and side scatter signals reflected changes in size and structure of yeast cells, in all treatments applied. When flow cytometry was interpreted alongside the scanning electron microscopy images, bimodal populations of yeast cells with low and high PI intensities were revealed and distinctive ‘doughnut’-shaped cell features observed in association with the microwave treatment only at tirage, that were not observed until 12 months wine ageing in older lees from the control. This work offers both a rapid approach to visualise alterations to yeast cell surfaces and a better understanding of the mechanisms of yeast lysis. Microwave, ultrasound or β-glucanase enzymes are tools that could potentially initiate the release of yeast cell compounds into wine. Further investigation into the impact of such treatments on the flavour and aroma profiles of the wines through sensory evaluation is warranted.

Premium sparkling wine produced by the traditional method (analogous to the French méthode champenoise) is characterised by the development of aged wine character as a result of a second fermentation in the bottle with lees contact and lengthy ageing. Treatments (microwave, ultrasound, or β-glucanase enzymes) were applied to disrupt the cell wall of Saccharomyces cerevisiae and added to the tirage liquor for the second fermentation of Chardonnay-Pinot Noir base wine cuvée and compared to a control, to assess effects on the release of phenolics, proteins, amino acids, and lipids at 6, 12 and 18 months post-tirage. General responses to wine ageing included a 60% increase in the total phenolic content of older sparkling wines relative to younger wines and an increase in protein concentration from 6 to 12 months bottle age. Microwave and β-glucanase enzyme treatments of yeast during tirage preparation were associated with a 10% increase in total free amino acid concentration and a 10% increase in proline concentration at 18 months bottle age, compared to control and ultrasound treatment. Furthermore, microwave treatment was associated with elevated asparagine content in wine at 18 months bottle age, relative to the control and the other wines. The β-glucanase enzyme and ultrasound treatments were associated with significant accumulation of total lipids, which were driven by 2-fold increases in the phospholipid and monoacylglycerol components in wine at 18 months bottle age and, furthermore, the microwave treatment was associated with elevated triacylglycerol at 18 months bottle age. This study demonstrates that the use of yeast treatments at the tirage stage of sparkling wine production presents an opportunity to manipulate wine composition.