Yeasts are unicellular fungi that harbour a large biodiversity of thousands of species, of which particularly ascomycetous yeasts are instrumental to human food and beverage production. There is already a large body of evidence showing that insects play an important role for yeast ecology, for their dispersal to new habitats and for breeding and overwintering opportunities. Here, we sought to investigate a potential role of the terrestrial snails Cepaea hortensis and C. nemoralis, which in Europe are often found in association with human settlements and gardens, in yeast ecology. Surprisingly, even in a relatively limited culture-dependent sampling size of over 150 isolates, we found a variety of yeast genera, including species frequently isolated from grape must such as Hanseniaspora, Metschnikowia, Meyerozyma and Pichia in snail excrements. We typed the isolates using standard ITS-PCR-sequencing, sequenced the genomes of three non-conventional yeasts H. uvarum, Meyerozyma guilliermondii and P. kudriavzevii and characterized the fermentation performance of these three strains in grape must highlighting their potential to contribute to novel beverage fermentations. Aggravatingly, however, we also retrieved several human fungal pathogen isolates from snail excrements belonging to the Candida clade, namely Ca. glabrata and Ca. lusitaniae. Overall, our results indicate that diverse yeasts can utilise snails as taxis for dispersal. This courier service may be largely non-selective and thus depend on the diet available to the snails.
Kveik are consortia of yeast used for farmhouse ale production in Western Norway. Yeast strains derived from these mixtures are known, for example, for their high fermentation rate, thermotolerance, lack of phenolic off flavor production (POF-) and strong flocculation phenotype. In this study, we used five single cell yeast isolates from different Kveik yeasts, analyzed their fermentation and flavor production, and compared it with a typical yeast used in distilleries using 20 °C and 28 °C as the fermentation temperatures. One of the isolates, Kveik No 3, showed an impairment of maltotriose utilization and thus a reduced ethanol yield. Kveik fermentations for spirit production often harbor bacteria for flavor enrichment. We sought to improve Kveik fermentations with non-conventional yeasts (NCY). To this end we co-fermented Kveik isolates with Hanseniaspora uvarum, Meyerozyma guilliermondii and Pichia kudriavzevii using 5:1 ratios (Kveik vs. NCY) at 20 °C. The combinations of Kveik No 1 with P. kudriavzevii and Kveik No 1 with Hanseniaspora uvarum showed substantially increased amounts of specific volatile aroma compounds that were previously identified in the NCYs. Our results indicate that Kveik isolates appear to be suitable for co-fermentations with certain NCY to enhance beer or spirit fermentations, increasing the potential of these yeasts for beverage productions.
All laboratories dealing with microbes have to develop a strain maintenance regime. While lyophilization based on freeze-drying may be feasible for large stock centers, laboratories around the world rely on cryopreservation and freezing of stocks at −80 °C. Keeping stocks at these low temperatures requires investments of several thousand kW/h per year. We have kept yeast stocks for several decades at room temperature on agar slants in glass reagent tubes covered with vaspar and sealed with cotton plugs. They were part of the Geisenheim Yeast Breeding Center stock collection that was started in the 19th century, well before −80 °C refrigeration technology was invented. Of these stocks, 60 tubes were analyzed and around one-third of them could be regrown. The strains were typed by sequencing of rDNA PCR fragments. Based on BlastN analyses, twelve of the strains could be assigned to Saccharomyces cerevisiae, two to S. kudriavzevii, and the others to Meyerozyma and Candida. The strains were used in white wine fermentations and compared to standard wine yeasts Uvaferm/GHM (Geisenheim) and Lalvin EC1118. Even with added nitrogen, the strains exhibited diverse fermentation curves. Post-fermentation aroma analyses and the determination of residual sugar and organic acid concentrations indicated that some strains harbor interesting flavor characteristics, surpassing current standard yeast strains. Thus, old strain collections bear treasures for direct use either in wine fermentations or for incorporation in yeast breeding programs aimed at improving modern wine yeasts. Furthermore, this provides evidence that low-cost/long-term culture maintenance at zero-emission levels is feasible.
Lager yeasts are hybrids between Saccharomyces cerevisiae and S. eubayanus. Wine yeast biodiversity, however, has only recently been discovered to include besides pure S. cerevisiae strains also hybrids between different Saccharomyces yeasts as well as introgressions from non-Saccharomyces species. Here, we analysed the genome of the Champagne Epernay Geisenheim (CEG) wine yeast. This yeast is an allotetraploid (4n − 1) hybrid of S. cerevisiae harbouring a substantially reduced S. kudriavzevii genome contributing only 1/3 of a full genome equivalent. We identified a novel oligopeptide transporter gene, FOT4, in CEG located on chromosome XVI. FOT genes were originally derived from Torulaspora microellipsoides and FOT4 arose by non-allelic recombination between adjacent FOT1 and FOT2 genes. Fermentations of CEG in Riesling and Müller-Thurgau musts were compared with the S. cerevisiae Geisenheim wine yeast GHM, which does not carry FOT genes. At low temperature (10°C), CEG completed fermentations faster and produced increased levels of higher alcohols (e.g. isoamyl alcohol). At higher temperature (18°C), CEG produced higher amounts of the pineapple-like alkyl esters i-butyric and propionic acid ethyl esters compared to GHM. The hybrid nature of CEG thus provides advantages in grape must fermentations over S. cerevisiae wine yeasts, especially with regard to aroma production.
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