Search Results (97)

The transition towards a circular bioeconomy is essential to mitigate the environmental pressures caused by the increasing global demand for food and energy. Agro-food waste (AFW) is a plentiful, inexpensive feedstock, exploitable in biorefineries to produce valuable molecules. The aim of this study was to isolate native non-conventional yeasts (NCY) from various AFW and to evaluate their potential for the ‘natural’ synthesis of aroma compounds via fermentation. Ten strains were isolated and identified as belonging to Saccharomyces cerevisiae, Pichia kluyveri, Pichia californica and Wickerhamomyces anomalus species. The fermentative performance and production of aroma volatile compounds were tested using different household wastes as substrates. Figs containing substrate, which is the richest in fermentable sugars, allowed for the fastest microbial adaptation and highest yields of volatile compounds. HS-SPME-GC/MS analysis revealed that the most prominent compounds were isoamyl alcohol, ethyl acetate and isoamyl acetate with the highest production levels showed by W. anomalus YDSCYP4 and P. kluyveri YDSCYP5. Enzymatic profiling revealed significant arylamidase and esterase activities in the selected strains, related to their role in the hydrolysis of aroma precursors. These findings demonstrate the efficiency of these autochthonous yeasts for the sustainable production of aroma compounds, supporting the development of eco-friendly biotechnological processes. Full article

This study investigated the biochemical composition and antioxidant potential of flours from pigment-deficient Chlorella vulgaris variants (honey and white) and wild-type (green) and the impact of lactic acid bacteria–yeast co-culture fermentation. The three variants were characterized for composition, total polyphenol (TPC) and flavonoid (TFC) contents, antioxidant capacity (DPPH, FRAP, and ORAC assays), and reactive oxygen species production in HT-29 intestinal cells. All extracts were noncytotoxic up to 100 µg/mL. Among all variants, the green showed the highest native TPC, TFC, and overall antioxidant activity. TPC and TFC were similar between honey and white, while FRAP was higher in honey and ORAC was higher in white. Biomasses were subsequently fermented for 24 h using Lactiplantibacillus plantarum CR L1 or Levilactobacillus brevis L204 with either Saccharomyces cerevisiae TRE Y100 or Kluyveromyces marxianus MK Y55. Fermentation resulted in significant pH reduction and increases in titratable acidity and lactic acid production, particularly in co-cultures involving K. marxianus. However, the effects on antioxidant properties were strongly matrix-dependent, with significant increases in TPC and antioxidant activity observed only in the white variant. Overall, pigmentation and microbial pairing emerged as key determinants of metabolic outcomes. These findings highlight the potential of co-culture fermentation to enhance the bioactive profile of pigment-deficient C. vulgaris, supporting their application in functional foods. Full article

Pomegranate peel, accounting for 35–50% of the fruit weight, is an underutilized agri-food by-product. This study applied, for the first time, fermentation with Saccharomyces cerevisiae as a simple and sustainable strategy to simultaneously obtain tannin-rich extracts and polysaccharide fractions with potential prebiotic activity. Peels from two cultivars, Wonderful and G1, differing in peel thickness, were subjected to three fermentation protocols (air- and not air-exposed) and monitored at 25 °C over 48 and 72 h. HPLC-DAD analysis showed that yeast-inoculated fermentation increased total tannin concentration in dry extracts (up to 70%) without inducing chemical modifications to tannin profiles. As determined by Dynamic Light Scattering, fermentation promoted significant depolymerization of native polysaccharides, while DOSY-¹H-NMR analyses revealed the presence of reduced molecular weight fractions down to 26 kDa. In vitro growth assays confirmed that fermented polysaccharides were more efficiently utilized as a carbon source by Bifidobacterium breve and Lactiplantibacillus plantarum compared to non-fermented controls, likely thanks to polysaccharide depolymerization induced by fermentation. The study demonstrated that air-exposed S. cerevisiae fermentation was an effective process alternative to chemical or enzymatic hydrolysis for modifying pomegranate peel pectin directly within a complex matrix, while simultaneously enhancing tannin recovery. This approach represents a possible sustainable strategy for pomegranate peel valorization into functional ingredients. Full article

Traditional fermented beverages from Chiapas, Mexico, represent an important source of microbial diversity, particularly of Saccharomyces cerevisiae. In native strains isolated from traditional fermented beverages, Saccharomyces cerevisiae has been observed to display distinct morphological and physiological traits; therefore, the aim of this study was to evaluate the population growth and the tolerance of twenty isolates to different stress factors such as temperature, osmotic pressure, and high ethanol concentrations, as well as the genetic variability through interdelta analysis, and to determine whether these physiological and molecular characteristics are associated with the type of beverage and the locality of origin. Differences were observed in tolerance to various factors, including high ethanol concentrations and elevated temperatures, as well as in the production of volatile compounds, with Taberna and Mezcal isolates showing notable performance. These isolates were able to withstand temperatures ranging from 43 to 45 °C and ethanol concentrations of up to 17% in Mezcal and Pox isolates, and 15% in Taberna isolates. High concentrations of isoamyl acetate and higher alcohols such as isoamyl alcohol were detected. In addition, the genetic variability of the isolates was evaluated, and its relationship with the type of beverage and the geographical origin of production was explored, including isolates obtained from Taberna, Mezcal, Pox, and Chicha de Chilacayote. Intraspecific variability was assessed through a retrotransposon-based analysis of the interdelta region using different primer combinations (δ1-δ2, δ12-δ21, and δ12-δ2). The generated banding patterns were analyzed using the Unweighted Pair Group Method with Arithmetic Mean (UPGMA), which enabled the identification of molecular variability patterns among the isolates. Furthermore, a UPGMA analysis was performed using physiological and compound production data, revealing a relationship between these characteristics and the geographical origin of the isolates. The results revealed a high degree of intraspecific variability, which was associated with both the type of beverage and the locality of origin of the isolates. Full article

Sorghum (Sorghum bicolor (L.) Moench) is increasingly recognized as a sustainable crop due to its adaptability to challenging environmental conditions and its nutritional potential. The present study aimed to characterize the nutritional composition and native microbial species associated with three sorghum hybrids cultivated in Bulgaria. Crude protein was 9.37–10.42%, total carbohydrate content was between 87.4 and 89.6%, and crude fat content was in the range of 3.84–4.9%. Linoleic acid was the predominant fatty acid in all hybrids, accounting for 44.9% to 48.0% of total lipids. Quinic acid emerged as the dominant organic acid in all hybrids, with the highest concentration of 729.37 mg/100 g. The microbiological assessment focused on lactic acid bacteria (LAB) and yeasts. Microbial isolates were subjected to molecular identification through 16S rRNA gene and ITS region sequencing. The predominant LAB species included Levilactobacillus brevis, Lactiplantibacillus plantarum, Lactiplantibacillus pentosus, Pediococcus acidilactici, and Pediococcus pentosaceus, while most of the yeast isolates belonged to Saccharomyces cerevisiae. Phylogenetic analysis indicated substantial intraspecies variation, particularly within LAB strains, suggesting the presence of unique genotypic traits. These findings contribute to a better understanding of sorghum’s nutritional value and endogenous microbiota and open opportunities for developing sorghum-based functional products. Full article

In this study, we investigated the impact of two native strains of Starmerella bacillaris, used both in pure culture and in a co-inoculation with Saccharomyces cerevisiae, on the volatile profile of a chemically defined fermented model must. The focus of this study was the production of monoterpenes and sesquiterpenes and their potential sensory contributions. Geraniol and linalool were detected in all fermentations with Starmerella bacillaris, in ranges of 26.7–43.9 µg/L and 34.3–41.3 µg/L, respectively, independent of the inoculation strategy used. Both strains produced concentrations above their respective odour thresholds of 20 µg/L and 25.5 µg/L. Odour activity value (OAV) analysis confirmed that fermentations with Starmerella bacillaris, particularly under co-inoculation conditions, generated the highest OAVs for these monoterpenes. Citronellol was only detected in mixed fermentations, while nerolidol and farnesol isomers were produced in variable amounts, depending on the strain and inoculation strategy, at concentrations below the odour threshold. These findings demonstrate the ability of Starmerella bacillaris to facilitate de novo biosynthesis of linalool, geraniol, and sesquiterpenes during alcoholic fermentation—in the case of linalool and geraniol, at concentrations exceeding their respective odour thresholds—highlighting the biotechnological potential of these native strains to enhance aroma in wines, particularly those made from neutral grape varieties. Full article

This study explores the production of anhydrous ethanol from discarded fruits, aiming to determine optimal fermentation conditions and evaluate the feasibility of a green separation technology. Fermentation experiments were performed using juices from Psidium guajava (S1), Carica paapaya (S2), and mucilage residues of Coffea arabica (S3). All fermentations were carried out at a pH of 4.5 for 7 days in 1 L bioreactors. A full 2² factorial design was applied to evaluate the effects of two variables: yeast type (commercial Saccharomyces cerevisiae [CY] vs. native yeast [NY]) and temperature (21 °C vs. 30 °C). Higher ethanol concentrations were achieved with CY at 30 °C, yielding 6.79% ethanol for S3. A multi-criteria matrix prioritized coffee residues due to their high ethanol yield, biomass availability, and economic viability. The ethanol was dehydrated using a packed-bed bioadsorption system with crushed corn, which increased purity from 6.7% v/v to 98.9% v/v in two stages, while avoiding azeotropic limitations. Energy analysis revealed low specific consumption (3.68 MJ/kg), outperforming conventional distillation. The results of this study, obtained at operating temperatures of 30 °C and 21 °C, a pH of 4.5, and an operating time of 7 days in a 1L bioreactor, demonstrate ethanol concentrations of 6.79%, confirming the technical feasibility of using agricultural waste as a raw material and validating the efficiency of a bioadsorption-based dehydration system. These findings address the current gap in integrating green ethanol separation with low-cost agricultural residues and highlight a sustainable alternative for decentralized bioethanol production. Full article

Histone 3 lysine 4 methylation (H3K4me) is an evolutionarily conserved epigenetic marker associated with transcriptional activation, playing a crucial role in growth and development. In yeast, all forms of H3K4 methylation are catalyzed by the COMPASS complex. However, purifying endogenous COMPASS remains challenging due to its low abundance, compositional complexity, and structural instability, resulting in low yield, poor purity, and heterogeneity in isolated complexes. These technical limitations have impeded the structural elucidation of the intact COMPASS complex and contributed to inconsistencies in reported in vitro enzymatic activity, thereby limiting a comprehensive understanding of its functions. Here, we present an optimized tandem affinity purification strategy that enables the high-yield isolation of native COMPASS from Saccharomyces cerevisiae with >99% purity and intact subunit composition, as validated by biochemical analyses. Using recombinant nucleosomes as substrates, we systematically characterized its catalytic properties and found that endogenously purified COMPASS exhibited strict dependence on H2B ubiquitination for catalyzing H3K4 methylation. This work establishes an efficient purification strategy for future structural and functional studies of COMPASS and provides critical insights into its catalytic properties. Full article

All traditional mezcal producers use artisan methods to produce mezcal. The low technological development in the elaboration processes results in low yield and high residual sugar concentration. First, this work optimized the concentration of initial sugars and yeast-assimilable nitrogen (YAN) in Agave durangensis juice fermentation at the laboratory level. A yield near 0.49 g EtOH/g sugar and a productivity of 1.54 g EtOH/L*h was obtained with an initial sugar concentration of 120 g/L and a YAN concentration of 0.227 g/L. Only Saccharomyces cerevisiae was found after 24 h of incubation at laboratory level, using MALDI-TOF identification. Agave durangensis heads crushed by the artisan process were used to test the inoculant performance. A mezcal yield of 11.6 kg agave/L of mezcal was obtained using the S cerevisiae inoculant and nitrogen addition, which was significantly different (p < 0.05) from other treatments. The population dynamics during fermentation were analyzed through isolation and identification using MALDI-TOF. Several yeast species (Pichia kluyveri, Torulaspora delbrueckii, Zygosaccharomyces bailii, and Saccharomyces cerevisiae) were found at the beginning of fermentation. Nonetheless, only S. cerevisiae was found at the end of fermentation. The implantation of the inoculant used was confirmed through the comparative analysis of amplification patterns of the GTG5 microsatellite of the strains identified as S. cerevisiae, finding that the inoculated strain proportion was greater than 80% of the yeast population. A technological alternative to increase the efficiency of the process is combining the addition of YAN and the inoculation of the native S. cerevisiae, which was isolated from artisan alcoholic fermentation of agave to produce mezcal. Full article

The introduction of heterologous pathways into microbial cell compartments offers several potential advantages, including increasing enzyme concentrations and reducing competition with native pathways, making this approach attractive for producing complex metabolites like fatty acids and fatty alcohols. However, measuring subcellular concentrations of these metabolites remains technically challenging. Here, we explored 3-hydroxypropionic acid (3-HP), readily quantifiable and sharing the same precursors—acetyl-CoA, NADPH, and ATP—with the above-mentioned products, as a reporter metabolite for peroxisomal engineering in the yeast Komagataella phaffii. To this end, the malonyl-CoA reductase pathway for 3-HP production was targeted into the peroxisome of K. phaffii using the PTS1-tagging system, and further tested with different carbon sources. Thereafter, we used compartmentalized 3-HP production as a reporter system to showcase the impact of different strategies aimed at enhancing the peroxisomal NADPH pool. Co-overexpression of genes encoding a NADPH-dependent redox shuttle from Saccharomyces cerevisiae (IDP2/IDP3) significantly increased 3-HP yields across all substrates, whereas peroxisomal targeting of the S. cerevisiae NADH kinase Pos5 failed to improve 3-HP production. This study highlights the potential of using peroxisomal 3-HP production as a biosensor for evaluating peroxisomal acetyl-CoA and NAPDH availability by simply quantifying 3-HP, demonstrating its potential for peroxisome-based metabolic engineering in yeast. Full article

Copper is an essential micronutrient and the ability to scavenge tightly bound or trace levels of copper ions at the host–pathogen interface is vital for fungal proliferation in animal hosts. Recent studies suggest that trace metal ion acquisition is critical for the establishment and propagation of Pseudogymnoascus destructans, the fungal pathogen responsible for white-nose syndrome (WNS), on their bat host. However, little is known about these metal acquisition pathways in P. destructans. In this study, we report the characterization of the P. destructans high-affinity copper transporter VC83_00191 (PdCTR1a), which is implicated as a virulence factor associated with the WNS disease state. Using Saccharomyces cerevisiae as a recombinant expression host, we find that PdCTR1a can efficiently traffic Cu ions into the yeast cytoplasm. Complementary studies in the native P. destructans fungus provide evidence that PdCTR1a transcripts and protein levels are dictated by Cu-bioavailability in the growth media. Our study demonstrates that PdCTR1a is a functional high-affinity copper transporter and is relevant to Cu homeostasis pathways in P. destructans. Full article

Yeast-yeast interactions have been studied mainly using wine yeasts. However, studies are rare for native yeasts from agave juice fermentation. Therefore, this work used strains isolated from the alcoholic fermentation of agave to study the survival of non-Saccharomyces yeasts co-cultivated with Saccharomyces cerevisiae in media of different nutritional qualities. First, the feasibility of using simple and low-cost culture media was demonstrated to study the interactions between Saccharomyces cerevisiae and non-Saccharomyces yeasts. The results presented here demonstrated the antagonistic effect exerted by S. cerevisiae on Torulaspora delbrueckii, which showed a more significant loss of viability. However, the nutritional composition of the culture medium also influences this effect. It was clear that a nutritionally rich medium improved the survival of non-Saccharomyces yeasts. Lastly, the change in the survival of non-Saccharomyces yeasts also entails a variation in the concentration and diversity of minor volatile compounds produced during fermentation. This was observed in the variety and relative abundance of compounds belonging to the most numerous chemical families, such as alcohols, esters, and terpenes. 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

The Brazilian Cerrado region has a rich plant diversity, with fruits that have peculiar and unique sensory characteristics. For these reasons, using these fruits for biotechnological production is a promising alternative, mainly to protect this biome from deforestation and degradation. The production of fermented acetic acid is an option to add value to native fruits and offer the market beverages with better nutritional quality and bioactive compounds. This work aimed to characterize fruits and to develop cagaita (Eugenia dysenterica DC.) acetic fermented beverage. The fruits were subjected to physical-chemical analyses in the first part. Subsequently, different treatments for fermentation were tested using two types of enzymes (amylase and pectinase), two subspecies of Saccharomyces cerevisiae yeast (UFLA CA11 and thermoresistant LNF Angel), and the chaptalization of the must with sucrose (16 °Brix). Alcoholic fermentation was carried out in an incubator with temperature control at 34 ± 1 °C. The pH, total soluble solids, titratable acidity, alcohol content, and density of the fermented products were monitored daily. The chaptalized must with amylase addition and thermoresistant yeast had the best performance during alcoholic fermentation, demonstrating that thermoresistant yeast is an economically advantageous and efficient alternative for the cagaita juice fermentation process. Subsequently, acetic fermentation was carried out using the slow method. Heat-resistant yeast without added enzymes was used to produce cagaita acetic fermented beverages within the parameters of the Brazilian legislation. Furthermore, phenolic compounds and antioxidant activity in the final product were observed. The work demonstrated the possibility of using cagaita fruits in biotechnological processes to produce new food products. Full article

In view of climate change and the increasingly antagonistic wine market, the exploitation of native genetic resources is revisited in relation to sustainable wine production. ‘Sideritis’ is a late-ripening Greek grape variety, which is quite promising for counteracting wine quality issues associated with the annual temperature rise. The aim of this study was to improve the quality and to enhance the aroma of ‘Sideritis’ wine through the use of native yeasts. To improve vinification, Hanseniaspora opuntiae L1 was used along with Saccharomyces cerevisiae W7 in mixed fermentations (SQ). The addition of H. οpuntiae significantly altered the chemical profile of the wine compared to the single-inoculated fermentations with W7 (IS). H. opuntiae increased all the acetate esters, except for hexyl acetate and (Z)-3-hexen-1-ol acetate. The concentration of 2-phenylethyl acetate, which imparts flowery and sweet notes, exhibited a 2.6-fold increase in SQ as compared to IS wines. SQ also showed higher levels in several ethyl esters, including ethyl butyrate, ethyl heptanoate and ethyl 7-octenoate, which are associated with fruity notes compared to IS. H. opuntiae produced citronellol, a terpene associated with rose and green notes, and increased the overall acceptance of the wine. Present results are thus quite promising for improving ‘Sideritis’ wine quality towards a sustainable wine production in Greece in view of global warming. Full article