Abstract
There is an increasing trend toward understanding the impact of non-Saccharomyces yeasts on the winemaking process. Although Saccharomyces cerevisiae is the predominant species at the end of fermentation, it has been recognized that the presence of non-Saccharomyces species during alcoholic fermentation can produce an improvement in the quality and complexity of the final wines. A previous work was developed for selecting the best combinations between S. cerevisiae and five non-Saccharomyces (Torulaspora delbrueckii, Schizosaccharomyces pombe, Candida stellata, Metschnikowia pulcherrima, and Lachancea thermotolorans) native yeast strains from D.O. “Vinos de Madrid” at the laboratory scale. The best inoculation strategies between S. cerevisiae and non-Saccharomyces strains were chosen to analyze, by real-time quantitative PCR (qPCR) combined with the use of specific primers, the dynamics of inoculated populations throughout the fermentation process at the pilot scale using the Malvar white grape variety. The efficiency of the qPCR system was verified independently of the samples matrix, founding the inoculated yeast species throughout alcoholic fermentation. Finally, we can validate the positive effect of selected co-cultures in the Malvar wine quality, highlighting the sequential cultures of T. delbrueckii CLI 918/S. cerevisiae CLI 889 and C. stellata CLI 920/S. cerevisiae CLI 889 and, mixed and sequential cultures of L. thermotolerans 9-6C combined with S. cerevisiae CLI 889.
Highlights
Alcoholic fermentation is a complex ecological and biochemical process where a succession of yeasts of several genera and species are able to convert must sugars into ethanol and carbon dioxide, as well as into important secondary metabolites (Barata et al, 2012; Sun et al, 2014; Albergaria and Arneborg, 2016)
The non-Saccharomyces strains used in this study are T. delbrueckii CLI 918, S. pombe CLI 1079, C. stellata CLI 920, M. pulcherrima CLI 457 and L. thermotolerans 9-6C, and S. cerevisiae CLI 889 strain were previously isolated on the Madrid winegrowing region and selected and characterized in our laboratories based on some established and desirable oenological criteria (Arroyo, 2000; Cordero-Bueso et al, 2013, 2016)
Conventional PCR was performed using purified DNA from the yeast species used in this study and different strains belonging to the yeasts species Candida vini, Wickerhamomyces anomalus, Zygosaccharomyces bailii, Meyerozyma guilliermondii, Pichia membranifaciens, Priceomyces carsonii, and Lachancea fermentati, the most usual species isolated during spontaneous fermentation of Malvar must in the experimental cellar of IMIDRA (Cordero-Bueso et al, 2013), which are included in the IMIDRA Institute Collection
Summary
Alcoholic fermentation is a complex ecological and biochemical process where a succession of yeasts of several genera and species are able to convert must sugars into ethanol and carbon dioxide, as well as into important secondary metabolites (Barata et al, 2012; Sun et al, 2014; Albergaria and Arneborg, 2016). Even though Saccharomyces species are present at a low frequency on the surface of healthy grapes, Saccharomyces cerevisiae is considered the primary microorganism in the fermentation process and it is widely used in oenology (Martini et al, 1996; Fleet, 2003). Yeast Monitoring on Mixed Fermentations during the last decade, non-Saccharomyces yeasts species have been proposed for winemaking as they could contribute to the improvement of wine quality (Ciani et al, 2014; Wang et al, 2015; Masneuf-Pomarede et al, 2016; Puertas et al, 2016). A new trend has emerged in winemaking using starter cultures composed by non-Saccharomyces yeasts, together with S. cerevisiae or for sequential fermentation with S. cerevisiae
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