Effect of non-wine Saccharomyces yeasts and bottle aging on the release and generation of aromas in semi-synthetic Tempranillo wines.

Interest in the use of non-conventional yeasts in wine fermentation has been increased in the last years in the wine sector. The main objective of this manuscript was to explore the aromatic diversity produced by wild and non-wine strains of S. cerevisiae, S. eubayanus, S. kudriavzevii, and S. uvarum species in young and bottle-aged Tempranillo wines as well as evaluate their fermentation capacity and the yield on ethanol, glycerol, and organic acids, that can contribute to diminishing the effects of climate change on wines. S. uvarum strain U1 showed the highest ability to release or de novo produce monoterpenes, such as geraniol and citronellol, whose values were 1.5 and 3.5-fold higher than those of the wine S. cerevisiae strain. We found that compared to the normal values for red wines, ß-phenylethyl acetate was highly synthesized by U1 and E1 strains, achieving 1 mg/L. Additionally, after aging, wines of S. eubayanus strains contained the highest levels of this acetate. Malic acid was highly degraded by S. kudriavzevii yeasts, resulting in the highest yields of lactic acid (>5-fold) and ethyl lactate (>2.8-fold) in their wines. In aged wines, we observed that the modulating effects of yeast strain were very high in ß-ionone. S. uvarum strains U1 and BMV58 produced an important aging attribute, ethyl isobutyrate, which was highly enhanced during the aging. Also, the agave S. cerevisiae strain develops an essential aroma after aging, reaching the highest ethyl leucate contents. According to the results obtained, the use of wild non-wine strains of S. cerevisiae and strains of the cryotolerant species S. eubayanus, S. kudriavzevii, and S. uvarum in Tempranillo wine fermentation increase the aroma complexity. In addition, wines from S. kudriavzevii strains had twice additional glycerol, those from S. uvarum 4-fold more succinic acid, while wines from wild strains yielded 1% v/v less ethanol which may solve wine problems associated with climate change.

GTR1 Affects Nitrogen Consumption and TORC1 Activity in Saccharomyces cerevisiae Under Fermentation Conditions.

The TORC1 pathway coordinates cell growth in response to nitrogen availability present in the medium, regulating genes related to nitrogen transport and metabolism. Therefore, the adaptation of Saccharomyces cerevisiae to changes in nitrogen availability implies variations in the activity of this signaling pathway. In this sense, variations in nitrogen detection and signaling pathway are one of the main causes of differences in nitrogen assimilation during alcoholic fermentation. Previously, we demonstrated that allelic variants in the GTR1 gene underlying differences in ammonium and amino acids consumption between Wine/European (WE) and West African (WA) strains impact the expression of nitrogen transporters. The GTR1 gene encodes a GTPase that participates in the EGO complex responsible for TORC1 activation in response to amino acids availability. In this work, we assessed the role of the GTR1 gene on nitrogen consumption under fermentation conditions, using a high sugar concentration medium with nitrogen limitation and in the context of the WE and WA genetic backgrounds. The gtr1Δ mutant presented a reduced TORC1 activity and increased expression levels of nitrogen transporters, which in turn favored ammonium consumption, but decreased amino acid assimilation. Furthermore, to identify the SNPs responsible for differences in nitrogen consumption during alcoholic fermentation, we studied the polymorphisms present in the GTR1 gene. We carried out swapping experiments for the promoter and coding regions of GTR1 between the WE and WA strains. We observed that polymorphisms in the coding region of the WA GTR1 gene are relevant for TORC1 activity. Altogether, our results highlight the role of the GTR1 gene on nitrogen consumption in S. cerevisiae under fermentation conditions.

Growth and lipid production of Rhodotorula glutinis R4, in comparison to other oleaginous yeasts.

Some Rhodotorula spp. have been characterized as oleaginous yeasts. Under certain culture conditions they can accumulate neutral lipids, which are mainly triglycerides (TAG). Microbial TAG that can be used as raw material for biodiesel synthesis are attractive for the biofuel industry. In this study, the ability to synthesize lipids of Rhodotorula glutinis R4, isolated in Antarctica, was compared with eight strains belonging to the genera Rhodotorula and Yarrowia with the aim of proposing a novel source of oils for biodiesel synthesis. All strains were cultured under nitrogen (N) limiting conditions and an excess of carbon (C) in the culture medium. We found that yeasts accumulated between 9-48.9 % (w/w) of lipids. Among them, R. glutinis R4 showed the highest growth (14 g L-1, µmax 0,092 h-1) and lipid production (7 g L-1; 47 % w/w). Microbial oils produced by R. glutinis R4 are similar to vegetable oils, with 61 % of oleic acid, indicating that it is adequate for biodiesel synthesis. Our results demonstrate that biodiesel derived from R. glutinis R4 complies with international fuel standards ASTM D6751 and EN 14214. Therefore, this work demonstrates that Rhodotorula glutinis R4 is a novel and valuable source of microbial oils for biodiesel synthesis.