Managing ethanol and sensory compounds by non-Saccharomyces yeasts

Abstract

The pursuit of flavour and phenolic ripeness, augmented by climate warming and extreme weather events, often leads to excessive sugar accumulation in grapes translating to undesirably high ethanol content in wines. Other common characteristics of such grapes and wines are inadequate acidity and aroma profiles, which all together compromise the quality and marketability of the final product. To tackle these issues, research has devised a number of methods implemented across the entire grape and wine production chain. Among these, partial fermentations with non-Saccharomyces yeasts is of particular interest, as it represents an undemanding approach that can also impart ‘complexity’ and distinctness. However, the full potential of non-Saccharomyces yeasts in wine ethanol and flavour management remains elusive, and this work aimed to further explore it. Research has shown that mixed fermentations with non-Saccharomyces yeasts can lead to enhanced wine aroma and sensory properties, and albeit limited, the current range of non- Saccharomyces is indeed marketed for this purpose. The potential of commercially available (and thus readily implementable) non-Saccharomyces co-inocula was assessed in Shiraz fermentations at two maturity levels; earlier harvest (24 °Brix) and technical ripeness (29 °Brix). Eight yeast treatments trialled in pilot scale fermentations included sequential inoculations using three Torulaspora delbureckii strains, one strain each of Lachancea thermotolerans and Metschnikowia pulcherrima, a commercial blend of non-Saccharomyces and S. cerevisiae, and appropriate S. cerevisiae controls. Fermentation monitoring and comprehensive chemical and sensory analysis allowed for the comparison of the treatments. The results showed a pronounced matrix-derived modulation of wine profiles which was reflective of grape maturity levels. Within each harvest date, however, the yeast treatments had a significant impact on a range of compositional and wine sensory characters. At earlier harvest stage, certain non-Saccharomyces treatments, in particular T. delbrueckii, led to an increased wine sensory appeal (i.e. ‘floral’, ‘red fruit’, ‘aroma intensity’, ‘spice’) compared to the S. cerevisiae control (‘vegetal, ‘acidic’ and ‘bitter’). These treatments, however, were related to incomplete fermentations in higher ripeness conditions. Thus, some non-Saccharomyces yeast showed promise in enhancing the quality of wines produced from earlier harvested grapes and as such represent a complementary approach in managing wine ethanol concentrations. The following study addressed the selection of lower-ethanol producing non-Saccharomyces strain(s) for use in sequential cultures with Saccharomyces cerevisiae. Oenological performances of six M. pulcherrima strains were evaluated in fermentations with S. cerevisiae inoculated after seven days. The best-performing M. pulcherrima MP2 strain was further characterised in six sequential fermentations with different S. cerevisiae inoculation delays in both synthetic and white grape juice. The analysis of main metabolites, undertaken prior to sequential inoculations and upon fermentation completion, highlighted metabolic interactions and carbon sinks other than ethanol in mixed culture fermentations. Depending on the inoculation delay, MP2 white wines contained between 0.6% and 1.2% (v/v) less ethanol than the S. cerevisiae control, with even larger decreases achieved in the synthetic juice. The MP2 wines also had higher concentrations of glycerol and lower concentrations of acetic acid. The analysis of volatile compounds revealed compositional alterations arising from the S. cerevisiae inoculation delay, with increased acetate esters and higher alcohols detected in all analysed MP2 treatments. The concept of intra-specific variability was studied using L. thermotolerans as a model. This species harbours several metabolic traits that are of value in oenology, including lactic acid production, potential to decrease ethanol content and modulate flavour in wines. The relationships between 172 L. thermotolerans isolates, sourced from natural and anthropic habitats worldwide, were studied using a 14-microsatellite genotyping method. The resultant clustering revealed that the evolution of L. thermotolerans has been driven by the geography and ecological niche of the isolation sources. Isolates originating from anthropic, in particular oenological environments, were genetically close, thus suggesting domestication events within the species. The phenotypic performance of the strains, assessed using a number of agar platebased growth assays with different carbon sources and physicochemical conditions, provided further support for the observed clustering. To determine whether, and to what extent, L. thermotolerans strains differ in the traits of oenological importance, and harbour signatures of domestication and/or local divergence, 94 previously genotyped strains were trialled in Chardonnay fermentations. The strains and the genetic groups were compared for their fermentation performance, production of primary and secondary metabolites and pH modulation. The common traits of L. thermotolerans strains were their glucophilic character, relatively extensive fermentation ability, low production of acetic acid and formation of lactic acid, which significantly affected the pH of the wines. An untargeted analysis of volatile compounds revealed that 58 out of 90 volatiles were affected at an L. thermotolerans strain level. Besides the remarkable extent of intra-specific diversity, results confirmed the distinct phenotypic performance of L. thermotolerans genetic groups. These observations provide further support for the occurrence of domestication events and allopatric differentiation in L. thermotolerans population.