Screening and Identification of Yeasts from Fruits and Their Coculture for Cider Production

Coculturing non-Saccharomyces yeasts with Saccharomyces cerevisiae could enrich the aromatic complexity of alcoholic beverages during cider brewing. Therefore, the present study performed rapid strain screening via selective culture medium and aroma analysis and adopted a response surface methodology to optimize fermentation conditions to produce 2-phenylethyl acetate (PEA), which presents a rose and honey scent. The effects of coculturing yeasts on cider quality were evaluated through hedonic sensory analysis and the check-all-that-apply (CATA) method. Hanseniaspora vineae P5 and S. cerevisiae P1 produced ciders with high levels of PEA and 2-phenylethanol, respectively. The optimal fermentation process consisted of sequential inoculation with a 31 h delay between inoculations, followed by fermentation for 14.5 d at 18.7 °C, yielding 17.41 ± 0.51 mg/L of PEA, which was 4.6-fold higher than that obtained through the unoptimized fermentation process. Additionally, the CATA results revealed that the cider produced through coculturing was associated with descriptors such as “smooth taste”, “honey”, “pineapple”, and “fruity”, which can be attributed to the high ethyl acetate and PEA levels in the cider.

Impact of Starmerella bacillaris and Zygosaccharomyces bailii on ethanol reduction and Saccharomyces cerevisiae metabolism during mixed wine fermentations

The bulk of grape juice fermentation is carried out by the yeast Saccharomyces cerevisiae, but non-Saccharomyces yeasts can modulate many sensorial aspects of the final products in ways not well understood. In this study, some of such non-conventional yeasts were screened as mixed starter cultures in a fermentation defined medium in both simultaneous and sequential inoculations. One strain of Starmerella bacillaris and another of Zygosaccharomyces bailii were chosen by their distinct phenotypic footprint and their ability to reduce ethanol levels at the end of fermentation, particularly during simultaneous vinification. S. bacillaris losses viability strongly at the end of mixed fermentation, while Z. bailii remains viable until the end of vinification. Interestingly, for most non-Saccharomyces yeasts, simultaneous inoculation helps for survival at the end of fermentation compared to sequential inoculation. S. cerevisiae viability was unchanged by the presence of the either yeast. Characterization of both strains indicates that S. bacillaris behavior is overall more different from S. cerevisiae than Z. bailii. S. bacillaris has a less strict glucose repression mechanism and molecular markers like catabolite repression kinase Snf1 is quite different in size. Besides, S. cerevisiae transcriptome changes to a bigger degree in the presence of S. bacillaris than when inoculated with Z. bailii. S. bacillaris induces the translation machinery and repress vesicular transport. Both non-Saccharomyces yeast induce S. cerevisiae glycolytic genes, and that may be related to ethanol lowering, but there are specific aspects of carbon-related mechanisms between strains: Z. bailii presence increases the stress-related polysaccharides trehalose and glycogen while S. bacillaris induces gluconeogenesis genes.

Use of Lachancea thermotolerans for Biological vs. Chemical Acidification at Pilot-Scale in White Wines from Warm Areas

Climate change is affecting vineyards, resulting in grapes with a low acidity a high pH and sugar at harvest time. The most common procedure so far to improve the acidity and reduce the final pH of wines is to use tartaric acid, but wine can also be acidified microbiologically using Lachancea thermotolerans yeasts, a natural bio-tool that acidifies gradually during the first stage/days of fermentation. Two strains of L. thermotolerans were compared with one Saccharomyces cerevisiae at a pilot-scale under similar fermentation conditions and in duplicate. A sequential inoculation was performed on the third day for the non-Saccharomyces, producing only about 1 g/L of lactic acid, which was suitable for comparison with the Saccharomyces, to which 1.5 g/L of tartaric acid had been added to lower the final pH. The three fermentations ended with a total acidity without significant differences. A significant and normal feature of the L. thermotolerans yeasts is their higher propane-1,2,3-triol production, which was observed in the Laktia yeast, and the acetic acid was <0.3 g/L. The amount of volatile metabolites was generally higher for non-Saccharomyces and the increase was seen in carbonyl compounds, organic acids, lactones, fumaric compounds, and phenols. Finally, the sensory analysis showed that there were hardly any significant differences, even though the non-Saccharomyces had a higher quantity of volatile metabolites, which could lead to a good acceptance of the product, since biological acidification was used, generating a more natural product.

Survey of Inoculated Commercial Saccharomyces cerevisiae in Winery-Based Trials

Wine production has developed from spontaneous to controlled fermentations using commercial active dry yeasts (ADY). In this study, S. cerevisiae commercial ADY were tested, and yeast community dynamics were monitored at different fermentation stages in three winery-based trials with volumes ranging from 60 L to 250 hL. The differentiation of S. cerevisiae strains was achieved using microsatellite markers. In Experiment 1, results showed that both ADY strains revealed similar profiles, despite being described by the producer as having different properties. In Experiment 2, higher genetic diversity was detected when co-inoculation was tested, while in sequential inoculation, the initial ADY seemed to dominate throughout all fermentation. Pilot-scale red wine fermentations were performed in Experiment 3, where one single ADY strain was tested along with different oenological additives. Surprisingly, these trials showed an increase in distinct profiles towards the end of fermentation, indicating that the dominance of the ADY was lower than in the blank modality. The use of ADY is envisaged to promote a controlled and efficient alcoholic fermentation, and their purchase represents an important cost for wineries. Therefore, it is most relevant to survey commercial ADY during wine fermentation to understand if their use is effective.

Host preference and invasiveness of commensal bacteria in the Lotus and Arabidopsis root microbiota

Roots of different plant species are colonized by bacterial communities, that are distinct even when hosts share the same habitat. It remains unclear to what extent the host actively selects these communities and whether commensals are adapted to a specific plant species. To address this question, we assembled a sequence-indexed bacterial culture collection from roots and nodules of Lotus japonicus that contains representatives of most species previously identified using metagenomics. We analysed taxonomically paired synthetic communities from L. japonicus and Arabidopsis thaliana in a multi-species gnotobiotic system and detected signatures of host preference among commensal bacteria in a community context, but not in mono-associations. Sequential inoculation experiments revealed priority effects during root microbiota assembly, where established communities are resilient to invasion by latecomers, and that host preference of commensal bacteria confers a competitive advantage in their cognate host. Our findings show that host preference in commensal bacteria from diverse taxonomic groups is associated with their invasiveness into standing root-associated communities.

Chemical and Sensory Profiles of Merlot Wines Produced by Sequential Inoculation of Metschnikowia pulcherrima or Meyerzyma guilliermondii

Inoculation of selected non-Saccharomyces yeasts with Saccharomyces cerevisiae as means to produce Merlot wines with reduced ethanol contents was investigated. Fermentations of grape musts (25.4° Brix, pH 3.50, and 4.23 g/L titratable acidity) were conducted in stainless steel tanks inoculated with Metschnikowia pulcherrima strains P01A016 or NS-MP or Meyerozyma guilliermondii P40D002 with S. cerevisiae Syrah added after three days. After fermentation, wines with Mt. pulcherrima contained 13.8% (P01A016) or 13.9% (NS-MP) v/v ethanol, respectively, amounts which were lower than in wines with S. cerevisiae alone (14.9% v/v). Delayed inoculation of must with S. cerevisiae (day 3) or musts with My. guilliermondii contained elevated concentrations of ethyl acetate (145 and 148 mg/L, respectively), concentrations significantly higher than those with S. cerevisiae inoculated on day 0 or with either strain of Mt. pulcherrima. Descriptive sensory analysis revealed a significant effect due to panelist but not due to Mt. pulcherrima or My. guilliermondii. This research indicates the potential for commercial application of these yeasts towards the production of reduced alcohol wines but without imparting negative sensory attributes.

Impact of Lachancea thermotolerans strain and lactic acid concentration on Oenococcus oeni and malolactic fermentation in wine

The yeast Lachancea thermotolerans can produce lactic acid during alcoholic fermentation (AF) and thereby acidify wines with insufficient acidity. However, little is known about the impact of L. thermotolerans on Oenococcus oeni, the primary lactic acid bacterium used in malolactic fermentation (MLF). This study explored the impact of sequential cultures of L. thermotolerans and Saccharomyces cerevisiae on MLF performance in white and red wines. Four L. thermotolerans strains were tested in Sauvignon blanc with sequential S. cerevisiae inoculation, compared to an S. cerevisiae control and the initially un-inoculated treatments. The L. thermotolerans wines showed large differences in acidification, and progression of MLF depended on lactic acid production, even at controlled pH. The highest and lowest lactic acid producing strains were tested further in Merlot fermentations with both co-inoculated and sequentially inoculated O. oeni. The low lactic acid producing strain enabled successful MLF, even when this failed in the S. cerevisiae treatment, with dramatically quicker malic acid depletion in O. oeni co-inoculation than in sequential inoculation. In contrast, a high lactic acid producing strain inhibited MLF irrespective of the O. oeni inoculation strategy. In a follow-up experiment, increasing concentrations of exogenously added lactic acid slowed MLF and reduced O. oeni growth across different matrices, with 6 g/L of lactic acid completely inhibiting MLF. The results confirm the inhibitory effect of lactic acid on O. oeni while highlighting the potential of some L. thermotolerans strains to promote MLF and the others to inhibit it.

Effect of Individual, Simultaneous and Sequential Inoculation of Pseudomonas fluorescens and Meloidogyne incognita on Growth, Biochemical, Enzymatic and Nonenzymatic Antioxidants of Tomato (Solanum lycopersicum L.)

This study was conducted on tomato (Solanum lycopersicum cv. K-21) to investigate the bioprotective nature of Pseudomonas fluorescens and its interactive effects with Meloidogyne incognita in terms of growth biomarkers, changes in biochemical attributes and modulation in antioxidant enzymes of the tomato plant. In this study, we grew tomato plants with M. incognita and P. fluorescens in separate pots, simultaneously and sequentially (15 days prior or post) after 15 days of seed sowing. The sequential inoculation of Mi15→Pf maximally increased the root-knot index and decreased the nematode population. It was also noted that inoculation suppressed the plant growth biomarkers in comparison to control. However, maximum suppression in nematode reproduction and increment in growth and physiological attributes were observed when P. fluorescens was applied 15 days prior to the nematode (Pf15→Mi) as compared to control. All the treatments showed an increase in antioxidant enzymes. Expression of phenol content and defensive enzymes such as peroxidase (POX) and superoxide dismutase (SOD) increased, in contrast to a significant reduction in malondialdehyde (MDA) and hydrogen peroxide (H2O2) contents when compared with the untreated inoculated plants. However, the highest levels of POX and SOD, and a lowest of phenol, MDA and H2O2 were displayed in the treatment Pf15→Mi, followed by Mi+Pf and Mi15→Pf.