Search Results (35)

Over the past few years, consumers’ preference for wines with a distinct regional identity has shifted producers to adopt traditional grape varieties and has increased interest in the indigenous grapevine microbiome, due to its potential influence on the wine quality characteristics. In the present study, indigenous Saccharomyces cerevisiae strains were isolated from 3 local grape varieties (“Keratsouda”, “Karnachalas”, and “Bogialamas”) of the Soufli region in northern Greece. After molecular identification, all strains were evaluated for key oenological traits (ethanol resistance, SO₂ resistance, H₂S production, and aminoacid decarboxylation). Three representative strains (one per grape variety) were further evaluated for their long-term survival after freeze-drying and their fermentation potential at 12 °C and 22 °C. Ethanol content ranged in similar values (11.9–12.5%) for all wines, but kinetic parameters were significantly affected by the strain used and the fermentation temperature. Principal component analysis revealed that the volatile composition as well as practical wine attributes were affected by both the strain used and the fermentation temperature and resulted in distinct sensory profiles in the new products. Full article

Winemaking involves a microbial ecosystem where yeast diversity, shaped by terroir and winemaking conditions, determines wine characteristics. Understanding the microbial diversity of vineyards and spontaneous fermentation is crucial for explaining a winery’s typical wine profile. Studying and inoculating indigenous strains make it possible to produce high quality wines, reflecting the production environment. This study analyzes the yeast species involved in 16 spontaneous fermentations (8 in 2022 and 8 in 2023) from grapes of four distinct vineyards under two sets of winemaking conditions. A total of 1100 yeast colonies were identified by MALDI-TOF and DNA sequencing techniques. Saccharomyces (S.) cerevisiae and Hanseniaspora uvarum were the most prevalent species, alongside significant populations of non-Saccharomyces yeasts such as Lachancea thermotolerans and Metchnikowia pulcherrima, which were the most abundant ones. Minor yeast species, including Aureobasidium pullulans, Starmerella bacillaris, Kazachstania servazzi, and other Hanseniaspora spp., were also detected. The results demonstrated that yeast diversity in spontaneous fermentations varied according to vineyard origin and winemaking conditions. Differences between the two vintages studied indicated that annual climatic conditions significantly influenced yeast diversity, especially among non-Saccharomyces species. This substantial diversity represents a valuable source of indigenous yeasts for preserving the typicity of a winery’s wines under controlled conditions. Full article

Non-Saccharomyces yeasts have the potential to ameliorate wine ethanol levels, but such fit-for-purpose yeast strains are still lacking. Seventy-one indigenous non-Saccharomyces yeasts isolated from spontaneous fermentations of four wine regions in China (Ningxia, Xinjiang, Gansu, and Shaanxi) were screened for ethanol formation and were characterized for major metabolite profiles in synthetic grape juice fermentation to obtain non-Saccharomyces yeasts with low ethanol yields. Four Hanseniaspora strains with less volatile acidity production were primarily selected, and their ethanol yield was reduced by 22–32% compared to S. cerevisiae. These strains were further evaluated for oenological properties, namely ethanol and temperature tolerance, H₂S production, and killer activities against S. cerevisiae. Strain HuC-3-2 was then subjected to Atmospheric Room Temperature Plasma (ARTP) mutagenesis, and a mutant (HuC32-2-72) with rapid growth and optimized ethanol-reducing capability was obtained. The best-performing strains were further characterized in sequential fermentations with S. cerevisiae in Merlot juice, and resulted in a 1.4% v/v decrease in ethanol yield. Comprehensive analysis of yeast populations and the production of key metabolites highlighted important carbon sinks, as well as glycerol formation, partially accounting for the ethanol reduction. In addition to ethanol amelioration, the Hanseniaspora strains also led to alterations in many metabolites, including volatile compounds and some organic acids, which can further modulate wine aroma and flavor. Full article

The production of strawberry wine is an effective strategy for addressing the significant economic losses caused by strawberry spoilage. In recent years, there has been an increase in consumer demand for quality and flavor diversity in fruit wines. Therefore, it is necessary to develop novel strawberry wine products. In this research, we assessed and analyzed the influences of fermentation with Hanseniaspora uvarum, Kurtzmaniella quercitrusa, and Saccharomyces cerevisiae under four fermentation conditions on the fermentation kinetics, organoleptic characteristics, chemical compositions, antioxidant capacities, and flavor profiles of strawberry wines. Strawberry wines fermented with the indigenous non-Saccharomyces yeasts H. uvarum and K. quercitrusa showed higher 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-Azinobis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS) free-radical-scavenging capacities and significantly different flavor profiles compared to strawberry wines fermented with S. cerevisiae. In addition, adjusting the initial soluble solids contents of strawberry juices and fermentation temperatures positively affected the quality and flavor profiles of strawberry wines fermented with the H. uvarum and K. quercitrusa strains. Under the condition of 18 °C–20 °Brix, strawberry wine fermented with K. quercitrusa presented the highest antioxidant capacity, with enhanced flavor diversity and color intensity. It is worth noting that K. quercitrusa can be an alternative yeast for producing high-quality strawberry wine with a distinct floral aroma. Full article

Cocoa fermentation plays a key role in defining chocolate’s flavor, with yeasts being central to this process. This study aimed to explore intraspecific genetic diversity of major indigenous yeasts (i.e., Saccharomyces cerevisiae and Pichia kudriavzevii), and their potential interaction in the cocoa pulp environment. Their metabolic intraspecific diversity was characterized in synthetic cocoa pulp medium. Then, Saccharomyces cerevisiae, Pichia kudriavzevii, and other strains were introduced to each other to evaluate their potential negative interaction. Interesting strain associations were selected to further explore their interaction in synthetic cocoa pulp medium as well as real fresh cocoa pulp. From a fermentation campaign in Ivory Coast, a set of Saccharomyces (S.) cerevisiae and Pichia (P.) kudriavzevii strains were isolated from batches classified according to their chocolate quality (i.e., standard, intermediate, or premium chocolate). Less abundant species (i.e., Torulaspora franciscae, Kluyveromyces marxianus) were also isolated and tested for their potential negative interactions with S. cerevisiae and P. kudriavzevii. A set of strains were selected and cultured in single and in co-culture in a minimal cocoa pulp synthetic medium and in fresh cocoa pulp to highlight potential positive and/or negative interactions regarding fermentative aroma profile (i.e., higher alcohols, acetate esters, medium-chain fatty acids, and ethyl esters). The results highlighted the dominance of S. cerevisiae in fermentation kinetics and medium- to long-chain ester production, contrasted with P. kudriavzevii’s efficiency in short-chain ester synthesis. Intraspecific aroma profile variations can be pointed out. The co-cultures of P. kudriavzevii and S. cerevisiae strains isolated from the premium chocolate batch had a positive impact on the fermented pulp aroma profile. Negative interactions were observed with Torulaspora franciscae, which eliminated P. kudriavzevii’s aroma expression. Finally, the comparison of the data obtained for the minimal cocoa pulp synthetic medium compared to the cocoa pulp allowed us to draw conclusions about the use of synthetic media for studying cocoa fermentation. These findings emphasize the complex microbial interactions in cocoa fermentation that could shape future cocoa bean aroma. Full article

Japan has numerous vineyards with distinct geographical and climatic conditions. To the best of our knowledge, there is no comprehensive analysis of the diversity of yeasts associated with wine grapes from Japan. This study aimed to determine yeast diversity in wine grapes from four wine-producing regions in Japan and to evaluate the physicochemical characteristics of wines produced with indigenous Saccharomyces cerevisiae strains isolated from two regions. A total of 2648 strains were isolated from nine wine grape samples. MALDI-TOF MS and 26S rDNA sequence analyses revealed that the strains belonged to 21 non-Saccharomyces yeasts and 1 Saccharomyces yeast (S. cerevisiae). Non-Saccharomyces yeasts were found in high quantities and were highly distributed among the wine grape samples. Differences in the distribution of the identified yeast species were noted among the different wine grape varieties and regions. Indigenous S. cerevisiae strains of different genotypes from different regions exhibit distinct physiological traits. Our findings are expected to enhance our understanding of the local yeasts associated with Japanese vineyards and contribute to obtaining cultures that can provide region-specific organoleptic characteristics to local wines produced in Japan. Full article

The yeast Saccharomyces cerevisiae ensures successful fermentation in winemaking, although the persistent use of commercial strains lead to the loss of aroma complexity of wines. Hence, the research of indigenous S. cerevisiae with proper oenological features and well adapted to specific wine-growing areas become of great interest for winemakers. Here, 206 pure cultures of S. cerevisiae were isolated from two wineries during a two-year sampling campaign and bio-typed through interdelta sequences analyses with the aim to evaluate the occurrence and persistence of the S. cerevisiae wild population linked to each winery. Both wineries belong to the same Verdicchio DOC wine area (Castelli di Jesi), and never used commercial yeasts during fermentation. Results showed 19 different biotypes with a specific population of S. cerevisiae in each winery, without cross-contamination with each other and with commercial starter strains. Moreover, inside each winery a persistence of some dominant biotypes was observed over time (three biotypes in winery 1; 95% of isolates in the two years and one biotype in winery 2; 20% of isolates in the two years), indicating a sort of “winery-effect”. The evaluation of S. cerevisiae populations for the oenological characters by microfermentations showed a proper and well distinct aromatic imprinting on the resulted wines supporting the concept of “winery effect”. Full article

We used inter-delta typing (IDT) and MALDI-TOF profiling to characterize the genetic and phenotypic diversity of 45 commercially available winemaking Saccharomyces cerevisiae strains and 60 isolates from an organic winemaker from Waipara, New Zealand, as a stratified approach for predicting the commercial potential of indigenous isolates. A total of 35 IDTs were identified from the commercial strains, with another 17 novel types defined among the Waipara isolates. IDT 3 was a common type among strains associated with champagne production, and the only type in commercial strains also observed in indigenous isolates. MALDI-TOF MS also demonstrated its potential in S. cerevisiae typing, particularly when the high-mass region (m/z 2000–20,000) was used, with most indigenous strains from each of two fermentation systems distinguished. Furthermore, the comparison between commercial strains and indigenous isolates assigned to IDT 3 revealed a correlation between the low-mass data (m/z 500–4000) analysis and the recommended use of commercial winemaking strains. Both IDT and MALDI-TOF analyses offer useful insights into the genotypic and phenotypic diversity of S. cerevisiae, with MALDI-TOF offering potential advantages for the prediction of applications for novel, locally isolated strains that may be valuable for product development and diversification. Full article

Assyrtiko is a rare ancient grape variety of Greece, which is known to produce Protected Designation of Origin (PDO) Santorini white wines. Besides the famous character of the volcanic terroir, Assyrtiko of Santorini is also marked by a low pH value and sharp acidity. The aim of the present study was to apply a new inoculation procedure that modulates the fermentation process by maintaining the unique sensorial characteristics of Assyrtiko wines based on acidity. For this purpose, the Lachancea thermotolerans species, known for the formation of lactic acid, was tested in sequential fermentation with three different Saccharomyces cerevisiae strains. At the end of the fermentation process, implantation control for S. cerevisiae strains (interdelta sequence profile analysis) was performed, oenological parameters were determined according to the OIV protocols, and the volatile compounds produced were measured by gas chromatography–mass spectrometry (GC/MS). Finally, all produced wines were evaluated by quantitative descriptive analysis by two groups of experts; the Greek team of oenologists from Santorini Island specialized in Assyrtiko wines, and the French team of oenologists specialized in wine from Bordeaux. As expected, the inoculated strain was the one that dominated the fermentation process, but nine S. cerevisiae indigenous strains were also identified in the produced wines. Lachancea thermotolerans produced 1 g/L of lactic and also modulated the volatile profile of the wines independently of the S. cerevisiae strain used. The origin of the panelists played an important role in bringing up sensorial traits, such as acidity. Our results led to a new interesting application of L. thermotolerans for white wine production adapted to climate change claims. Full article

Saccharomyces cerevisiae is commonly used for the production of alcoholic beverages, including cider. In this study, we examined indigenous S. cerevisiae and S. uvarum strains, both species commonly found in cider from Hardanger (Norway), for their strain-specific abilities to produce volatile and non-volatile compounds. Small-scale fermentation of apple juice with 20 Saccharomyces strains was performed to evaluate their aroma-producing potential as a function of amino acids (AAs) and other physicochemical parameters under the same experimental conditions. After fermentation, sugars, organic acids, AAs, and biogenic amines (BAs) were quantified using the HPLC–UV/RI system. A new analytical method was developed for the simultaneous determination of nineteen AAs and four BAs in a single run using HPLC–UV with prior sample derivatization. Volatile compounds were determined using HS-SPME-GC-MS. Based on 54 parameters and after the removal of outliers, the nineteen strains were classified into four groups. In addition, we used PLS regression to establish a relationship between aroma compounds and predictor variables (AAs, BAs, organic acids, sugars, hydrogen sulfide (H₂S) production, CO₂ release) of all 19 strains tested. The results of the VIP show that the main predictor variables affecting the aroma compounds produced by the selected yeasts are 16, belonging mainly to AAs. Full article

The increasing demand for craft beer is driving the search for novel ale yeast cultures from brewing-related wild environments. The focus of bioprospecting for craft cultures is to identify feral yeasts suitable to imprint unique sensorial attributes onto the final product. Here, we integrated phylogenetic, genotypic, genetic, and metabolomic techniques to demonstrate that sour beer during aging in wooden barrels is a source of suitable craft ale yeast candidates. In contrast to the traditional lambic beer maturation phase, during the aging of sour-matured production-style beer, different biotypes of Saccharomyces cerevisiae dominated the cultivable in-house mycobiota, which were followed by Pichia membranifaciens, Brettanomyces bruxellensis, and Brettanomyces anomalus. In addition, three putative S. cerevisiae × Saccharomyces uvarum hybrids were identified. S. cerevisiae feral strains sporulated, produced viable monosporic progenies, and had the STA1 gene downstream as a full-length promoter. During hopped wort fermentation, four S. cerevisiae strains and the S. cerevisiae × S. uvarum hybrid WY213 exceeded non-Saccharomyces strains in fermentative rate and ethanol production except for P. membranifaciens WY122. This strain consumed maltose after a long lag phase, in contrast to the phenotypic profile described for the species. According to the STA1+ genotype, S. cerevisiae partially consumed dextrin. Among the volatile organic compounds (VOCs) produced by S. cerevisiae and the S. cerevisiae × S. uvarum hybrid, phenylethyl alcohol, which has a fruit-like aroma, was the most prevalent. In conclusion, the strains characterized here have relevant brewing properties and are exploitable as indigenous craft beer starters. Full article

The use of earthenware amphorae in winemaking can give wines unique attributes enhancing their typicity. Therefore, in this study, spontaneous and inoculated in-amphora fermentations of Trebbiano Toscano grape must were monitored to assess the Saccharomyces cerevisiae strains occurring in each fermentation as well as the chemical characteristics of the wines. Strain typing via Interdelta analyses pointed out that the commercial starters did not dominate, showing 24% and 13% implantation percentages, and that 20 indigenous strains were present at different percentages, ranging from 2 to 20%, in inoculated and spontaneous fermentations. The assessment of the technical characteristics of the indigenous strains via fermentations at lab and pilot scale (20 L amphorae) and the sensory analysis of the experimental wines allowed for the selection of two indigenous strains to be used as starter cultures in comparison to a commercial strain in 300-L-amphorae vinifications in the cellar. The observed fermentative performances and sensory analysis of the experimental wines highlighted that one indigenous S. cerevisiae strain dominated the process and conferred distinctive sensory characteristics to the Trebbiano Toscano wine, demonstrating its effectiveness in managing the in-amphora fermentations. In addition, the results demonstrated the ability of amphorae to protect the polyphenolic compounds from oxidation during wine ageing. Indeed, the concentration of both hydroxycinnamic acids and flavonols decreased, with an average reduction of 30% and 14%, respectively, while hydroxybenzoic acids remained unchanged. Full article

S. cerevisiae strains were isolated and identified from vineyards and the spontaneous fermentation must at the eastern foot of Helan Mountain in China, and their oenological properties and fermentation abilities were analyzed. From the total of 199 S. cerevisiae strains isolated and identified, 14 isolates (F4-13, F5-7, F5-9, F5-12, F5-18, F5-19, F5-21, F6-8, F6-23, F9-23, SXY-4, HT-10, ZXY-17, MXY-19) exhibited excellent tolerance to sugar, SO₂, and ethanol. Among the isolates, the strain F4-13 exhibited the better oenological properties, with low H₂S production (+), suitable flocculation ability (58.74%), and reducing-L-malic acid ability (49.07%), and generated high contents of polyphenol, anthocyanin, tannin, terpenes, and higher alcohols, which contributed to the improvement of the red fruity and floral traits of the wines. The obtained results provide a strategy for the selection of indigenous S. cerevisiae for wine fermentation to produce high-quality wine with regional characteristics. Full article

Immobilization of microbial cells for the production of industrially important enzymes has been reported to offer the advantages of recyclability, higher yields and cost effectiveness. The search for an appropriate matrix that is affordable and easy to prepare is a significant topic in microbial biotechnology. Here, an abundant type of agro-industrial waste—corncob—was utilized as an immobilization matrix for the production of xylanase from an indigenous yeast strain, Saccharomyces cerevisiae MK-157. This is the first report describing xylanase production from immobilized S. cerevisiae. To render the corncob matrix more porous, alkaline pretreatment was undertaken and yeast cells were immobilized on the matrix by cultivating at 30 °C for 48 h in Sabouraud dextrose broth. After incubation, the immobilized matrix was transferred to mineral salt medium containing 1% xylan and incubated at 30 °C for 24 h. Xylanase production was determined in cell-free culture supernatant and the matrix was recycled for up to seven cycles. Moreover, xylanase-mediated saccharification was carried out using sugarcane bagasse as a substrate and the release of reducing sugars was monitored. The results showed that the immobilized yeast produced 4.97 IU mL⁻¹ xylanase in the first production cycle, indicating a >tenfold increase compared to the free cells. Xylanase production further increased to its maximum levels (9.23 IU mL⁻¹) in the fourth production cycle. Nonetheless, the cells retained 100% productivity for up to seven cycles. The volumetric and specific productivity of xylanase were also the highest in the fourth cycle. Scanning electron microscopy images revealed the rough surface of the untreated corncob, which became more porous after alkaline pretreatment. Immobilized yeast cells were also visible on the corncob pieces. The saccharification of a natural resource—sugarcane bagasse—using xylanase preparation yielded 26 mg L⁻¹ of reducing sugars. Therefore, it can be concluded that yeast strains can yield sufficient quantities of xylanase, allowing possible biotechnological applications. Moreover, corncob can serve as a cost-effective matrix for industrially important yeast strains. Full article

Oenological yeasts play a critical role in the winemaking process. In this study, the biodiversity of the non-Saccharomyces yeast was analyzed and monitored using high-throughput sequencing and culture-dependent approaches. Oenological and fermentation characteristics of these native yeasts were further investigated. A total of 241 fungus species and 5 species of culturable non-Saccharomyces yeasts were detected using high-throughput sequencing and culture-dependent approaches, respectively. Five strains of aroma-producing yeasts (K4, K14, K19, K21, and K26) were isolated, and their growth characteristics, carbon source utilization, hydrogen sulfide production performance, and β-glucosidase activity were different. The oenological condition tolerances of most strains were lower than that of commercial S. cerevisiae X16. The co-inoculum of these strains and S. cerevisiae X16 regulated the volatile aroma characteristics of the fermented Korla fragrant pear (KFP) fruit wine, enriching and complicating the aroma flavor. Thus, the combined inoculation of these indigenous wine yeasts and S. cerevisiae has some application potential in the production of KFP wine. Full article