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20 pages, 6058 KiB  
Article
The GPI-Anchored Aspartyl Proteases Encoded by the YPS1 and YPS7 Genes of Candidozyma auris and Their Role Under Stress Conditions
by Alvaro Vidal-Montiel, Daniel Clark-Flores, Eulogio Valentín-Gómez, Juan Pedro Luna-Arias, Erika Rosales-Cruz, César Hernández-Rodríguez, Lourdes Villa-Tanaca and Margarita Juárez-Montiel
J. Fungi 2025, 11(8), 573; https://doi.org/10.3390/jof11080573 (registering DOI) - 1 Aug 2025
Abstract
Candidozyma auris is a multidrug-resistant, thermo- and osmotolerant yeast capable of persisting on biotic and abiotic surfaces, attributes likely linked to its cell wall composition. Here, seven putative genes encoding yapsins, aspartyl proteases GPI-anchored to the membrane or cell wall, were identified in [...] Read more.
Candidozyma auris is a multidrug-resistant, thermo- and osmotolerant yeast capable of persisting on biotic and abiotic surfaces, attributes likely linked to its cell wall composition. Here, seven putative genes encoding yapsins, aspartyl proteases GPI-anchored to the membrane or cell wall, were identified in the genomes of C. auris CJ97 and 20-1498, from clades III and IV, respectively. The C. auris YPS1 gene is orthologous to the SAP9 of C. albicans. The YPS7 gene is orthologous to YPS7 in C. glabrata and S. cerevisiae, so that they may share similar roles. An in silico analysis suggested an interaction between pepstatin and the catalytic domain of Yps1 and Yps7. Although this inhibitor, when combined with caffeine, had a subtle effect on the growth of C. auris, it induced alterations in the cell wall. CauYPS1 and CauYPS7 expression increased under nutrient starvation and NaCl, and at 42 °C. The transcriptome of the 20-1498 strain suggests that autophagy may play a role in thermal stress, probably degrading deleterious proteins or maintaining cell wall and vacuolar homeostasis. Therefore, CauYps1 and CauYps7 may play a role in the cell wall integrity of C. auris in stress conditions, and they could be a target of new antifungal or antivirulence agents. Full article
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22 pages, 2422 KiB  
Article
A Conserved N-Terminal Di-Arginine Motif Stabilizes Plant DGAT1 and Modulates Lipid Droplet Organization
by Somrutai Winichayakul, Hong Xue and Nick Roberts
Int. J. Mol. Sci. 2025, 26(15), 7406; https://doi.org/10.3390/ijms26157406 (registering DOI) - 31 Jul 2025
Abstract
Diacylglycerol-O-acyltransferase 1 (DGAT1, EC 2.3.1.20) is a pivotal enzyme in plant triacylglycerol (TAG) biosynthesis. Previous work identified conserved di-arginine (R) motifs (R-R, R-X-R, and R-X-X-R) in its N-terminal cytoplasmic acyl-CoA binding domain. To elucidate their functional significance, we engineered R-rich sequences in the [...] Read more.
Diacylglycerol-O-acyltransferase 1 (DGAT1, EC 2.3.1.20) is a pivotal enzyme in plant triacylglycerol (TAG) biosynthesis. Previous work identified conserved di-arginine (R) motifs (R-R, R-X-R, and R-X-X-R) in its N-terminal cytoplasmic acyl-CoA binding domain. To elucidate their functional significance, we engineered R-rich sequences in the N-termini of Tropaeolum majus and Zea mays DGAT1s. Comparative analysis with their respective non-mutant constructs showed that deleting or substituting R with glycine in the N-terminal region of DGAT1 markedly reduced lipid accumulation in both Camelina sativa seeds and Saccharomyces cerevisiae cells. Immunofluorescence imaging revealed co-localization of non-mutant and R-substituted DGAT1 with lipid droplets (LDs). However, disruption of an N-terminal di-R motif destabilizes DGAT1, alters LD organization, and impairs recombinant oleosin retention on LDs. Further evidence suggests that the di-R motif mediates DGAT1 retrieval from LDs to the endoplasmic reticulum (ER), implicating its role in dynamic LD–ER protein trafficking. These findings establish the conserved di-R motifs as important regulators of DGAT1 function and LD dynamics, offering insights for the engineering of oil content in diverse biological systems. Full article
(This article belongs to the Special Issue Modern Plant Cell Biotechnology: From Genes to Structure, 2nd Edition)
14 pages, 1948 KiB  
Article
Molecular Responses of Saccharomyces cerevisiae to Growth Under Conditions of Increasing Corn Syrup and Decreasing Molasses
by Binbin Chen, Yu Chyuan Heng, Sharifah Nora Ahmad Almunawar, Elvy Riani Wanjaya, Untzizu Elejalde and Sandra Kittelmann
Fermentation 2025, 11(8), 432; https://doi.org/10.3390/fermentation11080432 - 28 Jul 2025
Viewed by 163
Abstract
Molasses, a by-product of raw sugar production, is widely used as a cost-effective carbon and nutrient source for industrial fermentations, including the production of baker’s yeast (Saccharomyces cerevisiae). Due to the cost and limited availability of molasses, efforts have been made [...] Read more.
Molasses, a by-product of raw sugar production, is widely used as a cost-effective carbon and nutrient source for industrial fermentations, including the production of baker’s yeast (Saccharomyces cerevisiae). Due to the cost and limited availability of molasses, efforts have been made to replace molasses with cheaper and more readily available substrates such as corn syrup. However, the quality of dry yeast drops following the replacement of molasses with corn syrup, despite the same amount of total sugar being provided. Our understanding of how molasses replacement affects yeast physiology, especially during the dehydration step, is limited. Here, we examined changes in gene expression of a strain of baker’s yeast during fermentation with increasing corn syrup to molasses ratios at the transcriptomic level. Our findings revealed that the limited availability of the key metal ions copper, iron, and zinc, as well as sulfur from corn syrup (i) reduced their intracellular storage, (ii) impaired the synthesis of unsaturated fatty acids and ergosterol, as evidenced by the decreasing proportions of these important membrane components with higher proportions of corn syrup, and (iii) inactivated oxidative stress response enzymes. Taken together, the molecular and metabolic changes observed suggest a potential reduction in nutrient reserves for fermentation and a possible compromise in cell viability during the drying process, which may ultimately impact the quality of the final dry yeast product. These findings emphasize the importance of precise nutrient supplementation when substituting molasses with cheaper substrates. Full article
(This article belongs to the Section Yeast)
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14 pages, 3198 KiB  
Article
Small Nucleolar RNA from S. cerevisiae Binds to Phosphatidylinositol 4,5-Bisphosphate
by Irma A. Jiménez-Ramírez, Miguel A. Uc-Chuc, Luis Carlos Rodríguez Zapata and Enrique Castaño
Non-Coding RNA 2025, 11(4), 55; https://doi.org/10.3390/ncrna11040055 - 28 Jul 2025
Viewed by 161
Abstract
Background: snoRNAs have traditionally been known for their role as guides in post-transcriptional rRNA modifications. Previously, our research group identified several RNAs that may bind to PIP2 with LIPRNA-seq. Among them, snR191 stood out due to its potential specific interaction with this [...] Read more.
Background: snoRNAs have traditionally been known for their role as guides in post-transcriptional rRNA modifications. Previously, our research group identified several RNAs that may bind to PIP2 with LIPRNA-seq. Among them, snR191 stood out due to its potential specific interaction with this lipid, distinguishing itself from other snoRNAs. However, a detailed study is needed to define the molecular interactions between RNA and lipids, which remain unknown but may serve as a mechanism for transport or liquid–liquid phase separation. This study aimed to determine the interaction between a snoRNA called snR191 and PIP2. Method: A novel methodology for RNA-PIP2 interaction was carried out. Total RNA from Saccharomyces cerevisiae was incubated with PIP2-bound nitrocellulose membranes and RT-PCR reactions. We performed the prediction of snR191-PIP2 interaction by molecular docking and in silico mutations of snoR191. Results: From LIPRNA-seq analysis, we identified that PIP2-bound RNAs were significantly enriched in diverse biological processes, including transmembrane transport and redox functions. Our RNA-PIP2 interaction approach was successful. We demonstrated that snR191 specifically interacts with PIP2 in vitro. The elimination of DNA ensured that the interaction assay was RNA-specific, strengthening the robustness of the experiment. PIP2 was docked to snR191 in a stem–loop–stem motif. Six hydrogen bonds across four nucleotides mediated the PIP2-snR191 interaction. Finally, mutations in snR191 affected the structural folding. Conclusions: In this study, we demonstrate the effectiveness of a new methodology for determining RNA–lipid interactions, providing strong evidence for the specific interaction between snR191 and PIP2. Integrating biochemical and computational approaches has allowed us to understand the binding of these biomolecules. Therefore, this work significantly broadens our understanding of snR191-PIP2 interactions and opens new perspectives for further research. Full article
(This article belongs to the Section Long Non-Coding RNA)
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20 pages, 4658 KiB  
Article
Valorizing Carasau Bread Residue Through Sourdough Fermentation: From Bread Waste to Bread Taste
by Simonetta Fois, Valentina Tolu, Vanna Sanna, Antonio Loddo, Manuela Sanna, Piero Pasqualino Piu, Daniela Piras, Tonina Roggio and Pasquale Catzeddu
Microorganisms 2025, 13(8), 1745; https://doi.org/10.3390/microorganisms13081745 - 25 Jul 2025
Viewed by 164
Abstract
Surplus bread accounts for a significant proportion of food waste in many countries. The focus of this study was twofold: firstly, to investigate the use of carasau bread residue as a sourdough substrate, and secondly, to reuse this sourdough into a new carasau [...] Read more.
Surplus bread accounts for a significant proportion of food waste in many countries. The focus of this study was twofold: firstly, to investigate the use of carasau bread residue as a sourdough substrate, and secondly, to reuse this sourdough into a new carasau baking process. Selected lactic acid bacteria (Lactiplantibacillus plantarum) and yeast strains (Saccharomyces cerevisiae and Wickerhamomyces anomalus) were used to inoculate three substrates: bread residue (S1), bread residue supplemented with durum wheat middlings (S2), and semolina (S3). Sourdoughs were refreshed for five days by backslopping, and microbiological and physicochemical analyses were performed. Results indicated that incorporating wheat middlings into bread residue enhanced microbial performance, as evidence by a decrease in pH from 6.0 to around 4.5 compared to using bread residue alone as a substrate. Carasau bread produced with the sourdough derived from bread residue and wheat middlings exhibited comparable physicochemical properties to commercial baker’s yeast carasau bread, but had better sensory properties, scoring a mean acceptability of 7.0 versus 6.0 for baker’s yeast bread. These results show that bread residue supplemented with wheat middlings can serve as a sourdough substrate, allowing its reuse in the baking process to produce high-quality carasau bread and promote the circular economy. Full article
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26 pages, 2915 KiB  
Review
Recent Knowledge in the Application of Saccharomyces cerevisiae in Aquaculture: A Bibliometric and Narrative Review
by Elshafia Ali Hamid Mohammed, Béla Kovács and Károly Pál
Antibiotics 2025, 14(8), 736; https://doi.org/10.3390/antibiotics14080736 (registering DOI) - 22 Jul 2025
Viewed by 434
Abstract
Aquaculture is a key food production sector responsible for meeting the nutritional needs of a rapidly growing global population. However, the emergence of disease outbreaks has become a major challenge for the aquaculture industry, resulting in significant economic losses. The use of costly [...] Read more.
Aquaculture is a key food production sector responsible for meeting the nutritional needs of a rapidly growing global population. However, the emergence of disease outbreaks has become a major challenge for the aquaculture industry, resulting in significant economic losses. The use of costly and toxic antibiotics for treatment has a negative impact on the aquatic environment. Consequently, there has been a growing interest in probiotics as a non-antibiotic approach to manage disease outbreaks and improve fish performance. The use of the yeast Saccharomyces cerevisiae (SC) has shown remarkable benefits in aquaculture. In February 2025, a systematic search was conducted based on the Web of Science (WoS) database for the period 2015–2025 to identify relevant studies investigating the beneficial effects of SC in aquaculture. After searching on WoS, 466 documents were found and analyzed using R-bibliometric package for comprehensive analysis to identify research gap, trends, and distribution of global literature that focuses on SC in aquaculture. The most relevant and recent articles were reviewed, summarized and discussed. The yeast SC have shown a wide range of benefits, including improved growth performance, feed efficiency, enhanced diversity of the gut microbiome and immune response. The implementation of SC is becoming a recent trend and its efficacy in aquatic environments has been thoroughly investigated. This review aims to provide a valuable insight into SC as one of the most important aquaculture probiotics. It also emphasizes the need for further research to fully understand its benefits and the way it works. Full article
(This article belongs to the Special Issue Challenges and Strategies for the Antibiotic Resistance Crisis)
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11 pages, 1123 KiB  
Article
Biodiversity of Yeast Species Isolated During Spontaneous Fermentation: Influence of Grape Origin, Vinification Conditions, and Year of Study
by Ana Benito-Castellanos, Beatriz Larreina, María Teresa Calvo de La Banda, Pilar Santamaría, Lucía González-Arenzana and Ana Rosa Gutiérrez
Microorganisms 2025, 13(7), 1707; https://doi.org/10.3390/microorganisms13071707 - 21 Jul 2025
Viewed by 337
Abstract
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 [...] Read more.
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
(This article belongs to the Special Issue Wine Fermentation Microorganisms)
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26 pages, 2712 KiB  
Article
[1,3]Thiazolo[3,2-b][1,2,4]triazolium Salts as Effective Antimicrobial Agents: Synthesis, Biological Activity Evaluation, and Molecular Docking Studies
by Mykhailo Slivka, Boris Sharga, Daryna Pylypiv, Hanna Aleksyk, Nataliya Korol, Maksym Fizer, Olena I. Fedurcya, Oleksandr G. Pshenychnyi and Ruslan Mariychuk
Int. J. Mol. Sci. 2025, 26(14), 6845; https://doi.org/10.3390/ijms26146845 - 16 Jul 2025
Viewed by 379
Abstract
This study focuses on the search for new effective synthetic antimicrobial compounds as a tool against the widespread presence of microorganisms resistant to existing drugs. Five derivatives of [1,3]thiazolo[3,2-b][1,2,4]triazoles were synthesized using an accessible protocol based on electrophilic heterocyclization and were characterized using [...] Read more.
This study focuses on the search for new effective synthetic antimicrobial compounds as a tool against the widespread presence of microorganisms resistant to existing drugs. Five derivatives of [1,3]thiazolo[3,2-b][1,2,4]triazoles were synthesized using an accessible protocol based on electrophilic heterocyclization and were characterized using infrared (FTIR) and nuclear magnetic resonance (NMR) spectroscopies, and their in vitro antimicrobial and antifungal activities were evaluated using the agar plate diffusion method and the microdilution plate procedure. Both antibacterial (Gram-positive and Gram-negative) and antifungal activities were found for the examined samples. The minimum inhibitory concentration (MIC) varied from 0.97 to 250 µg/mL, and the minimum bactericidal concentration (MBC) from 1.95 to 500 µg/mL. Compound 2a showed good antifungal action against Candida albicans and Saccharomyces cerevisiae with minimum fungicidal concentration (MFC) 125 and MIC 31.25 µg/mL. The molecular docking revealed that the 2-heptyl-3-phenyl-6,6-trimethyl-5,6-dihydro-3H-[1,3]thiazolo[3,2-b][1,2,4]triazol-7-ium cation stands out as a highly promising candidate for further investigation due to a wide range of interactions, including conventional hydrogen bonds, π–σ, π–π T-shaped, and hydrophobic alkyl interactions. The synthesis and preliminary evaluation of [1,3]thiazolo[3,2-b][1,2,4]triazoles yielded promising antimicrobial and antifungal candidates. The diverse interaction profile of the 2-heptyl derivative salt allows this compound’s selection for further biological studies. Full article
(This article belongs to the Section Materials Science)
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15 pages, 725 KiB  
Article
In Vitro Evaluation of Ruminal Fermentation and Methane Production in Response to the Addition of Modified Nano-Bentonite with or Without Saccharomyces cerevisiae to a Forage-Based Diet
by Sohila Abo-Sherif, Sobhy Sallam, Ali M. Allam, Mounir El-Adawy and Yosra Soltan
Animals 2025, 15(14), 2081; https://doi.org/10.3390/ani15142081 - 15 Jul 2025
Viewed by 326
Abstract
Modified nano-clays, alone or combined with probiotics, may offer a novel and sustainable approach to improve ruminal fermentation and mitigate CH4 emissions in high-fiber diets. This study evaluated the properties and effects of modified nano-bentonite (MNB), with or without yeast (Saccharomyces [...] Read more.
Modified nano-clays, alone or combined with probiotics, may offer a novel and sustainable approach to improve ruminal fermentation and mitigate CH4 emissions in high-fiber diets. This study evaluated the properties and effects of modified nano-bentonite (MNB), with or without yeast (Saccharomyces cerevisiae), compared to natural bentonite (NB) and monensin, using the in vitro gas production (GP) technique. The substrate used was a basal diet composed primarily of forage (Trifolium alexandrinum clover) in a 70:30 forage-to-concentrate ratio. The treatments were a control group receiving the basal diet without additives; a monensin-added diet containing 40 mg/kg of dry matter (DM); a yeast-added diet with Saccharomyces cerevisiae at 2 × 108 CFU/g of DM; a NB clay-added diet at 5 g/kg of DM; and MNB diets added at two levels (0.5 g/kg of DM (MNBLow) and 1 g/kg of DM (MNBHigh)), with or without S. cerevisiae. MNB showed a smaller particle size and improved properties, such as higher conductivity, surface area, and cation exchange capacity, than NB. Sulfur and related functional groups were detected only in MNB. No differences were observed in total GP, while both the monensin diet and the MNBHigh-with-yeast diet significantly reduced CH4 emissions compared to the control (p < 0.05). The MNBHigh-without-yeast combination significantly (p < 0.05) reduced hemicellulose degradation, as well as total protozoal counts, including Isotricha and Epidinium spp. (p < 0.05), compared to the control. Ammonia levels did not differ significantly among treatments, while NB and MNBHigh diets tended to have (p = 0.063) the highest short-chain fatty acid (SCFA) concentrations. These findings suggest the potential modulatory effects of yeast and MNB on rumen fermentation dynamics and CH4 mitigation. Full article
(This article belongs to the Special Issue Feed Additives in Animal Nutrition)
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17 pages, 2439 KiB  
Article
CtGH76, a Glycoside Hydrolase 76 from Chaetomium thermophilum, with Elongated Glycan-Binding Canyon
by Silvana Ruth Ruppenthal, Wang Po-Hsun, Mohamed Watad, Christian Joshua Rosner, Marian Samuel Vogt, Markus Friedrich, Anna-Lisa Voigt, Angelique Petz, Petra Gnau and Lars-Oliver Essen
Int. J. Mol. Sci. 2025, 26(14), 6589; https://doi.org/10.3390/ijms26146589 - 9 Jul 2025
Viewed by 289
Abstract
Fungal cell walls, composed of polysaccharides and proteins, play critical roles in adaptation, cell division, and protection against environmental stress. Their polyglucan components are continuously remodeled by various types of glycosyl hydrolases (GHs) and transferases (GTs). In Saccharomyces cerevisiae and other ascomycetes, enzymes [...] Read more.
Fungal cell walls, composed of polysaccharides and proteins, play critical roles in adaptation, cell division, and protection against environmental stress. Their polyglucan components are continuously remodeled by various types of glycosyl hydrolases (GHs) and transferases (GTs). In Saccharomyces cerevisiae and other ascomycetes, enzymes of the Dfg5 subfamily, which belong as GTs to the GH76 family, cleave an α1,4 linkage between glucosamine and mannose to facilitate covalent linkage of GPI-anchored proteins to the cell wall’s polyglucans. In contrast, the functions of other fungal GH76 subfamilies are not understood. We characterized CtGH76 from the sordariomycete Chaetomium thermophilum, a member of the Fungi/Bacteria-mixed GH76 subfamily, revealing conserved structural features and functional divergence within the GH76 family. Notably, our structural characterization by X-ray crystallography combined with glycan fragment screening indicated that CtGH76 can recognize GPI-anchors like members of the Dfg5 subfamily but shows a broader promiscuity toward other glycans with central α1,6-mannobiose motifs due to the presence of an elongated glycan-binding canyon. These findings provide new insights into GH76 enzyme diversity and fungal cell wall maturation. Full article
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21 pages, 2551 KiB  
Article
SOD1 Deficiency Reveals Indirect Redox Stress Mechanisms Underlying Vanillin Toxicity in Saccharomyces cerevisiae Yeast
by Sabina Bednarska, Magdalena Kwolek-Mirek, Roman Maslanka, Dominika Graboś, Gabriela Świniuch and Renata Zadrag-Tecza
Antioxidants 2025, 14(7), 842; https://doi.org/10.3390/antiox14070842 - 9 Jul 2025
Viewed by 302
Abstract
Vanillin is a compound of great utility, and its production is, among others, based on using microorganisms such as Saccharomyces cerevisiae yeast. The effect of vanillin on cells is not fully understood. It has been demonstrated that vanillin induces oxidative stress; however, evidence [...] Read more.
Vanillin is a compound of great utility, and its production is, among others, based on using microorganisms such as Saccharomyces cerevisiae yeast. The effect of vanillin on cells is not fully understood. It has been demonstrated that vanillin induces oxidative stress; however, evidence also suggests its beneficial effects, including antioxidant and anti-inflammatory properties. For this reason, the present study was designed to elucidate the mechanism of vanillin’s action and to ascertain the extent to which its toxic effect is attributable to oxidative stress. The studies were conducted using wild-type and Δsod1 mutant strains. SOD1 deficiency results in cell hypersensitivity to oxidative factors, thus making the mutant strain a valuable model for investigating various aspects of oxidative stress. Based on an evaluation of cell vitality, Yap1p activation, ROS content, and glutathione and NADP(H) content, it can be concluded that oxidative stress is a secondary effect of metabolic and redox perturbations in cells rather than a direct consequence of vanillin reactivity. Furthermore, alterations observed in the redox couples GSH/GSSG and NADPH/NADP+ are one of the reasons for oxidative stress and suggest that vanillin may induce the utilization of NADPH for cellular needs other than antioxidant effects. Full article
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21 pages, 4028 KiB  
Article
The Response Characteristics of One Saccharomyces cerevisiae Strain Under Continuous Passage in Artificial Culture Medium
by Tengyu Ma, Hongguang Zhu, Jiajia Yin, Yu Tian, Wenjing Yan and Haixin Sun
J. Fungi 2025, 11(7), 513; https://doi.org/10.3390/jof11070513 - 9 Jul 2025
Viewed by 493
Abstract
Saccharomyces cerevisiae often undergoes strain degeneration during industrial serial subculturing, though this phenomenon remains understudied. This study first conducted strain screening and biological characterization through TTC (2,3,5-triphenyltetrazolium chloride) colorimetric assays, Durham tube fermentation gas production tests, and WL medium (Wallerstein Laboratory medium) cultivation. [...] Read more.
Saccharomyces cerevisiae often undergoes strain degeneration during industrial serial subculturing, though this phenomenon remains understudied. This study first conducted strain screening and biological characterization through TTC (2,3,5-triphenyltetrazolium chloride) colorimetric assays, Durham tube fermentation gas production tests, and WL medium (Wallerstein Laboratory medium) cultivation. Subsequently, the changes in intergenerational biological traits after serial subculturing were investigated. Finally, transcriptomic analysis was employed to examine differential gene expression under high-glucose stress during continuous subculturing. The experimental results demonstrated that: (1) The S. cerevisiae QDSK310-Z-07 (GenBank: PP663884), isolated from farm soil, exhibited robust growth within a temperature range of 24–36 °C, with optimal growth observed at 28 °C. It thrived in a pH range of 4–5.5 and efficiently utilized various carbon and nitrogen sources; (2) After serial subculturing, the strain’s ethanol production capacity and fermentation rate partially declined and then stabilized, while maintaining strong tolerance to high ethanol concentrations and hyperosmotic stress; (3) Transcriptomic analysis revealed significant differential expression of genes related to lipid metabolism, amino acid metabolism, and other pathways under high-glucose stress following continuous subculturing. These findings elucidate the biological trait variations in S. cerevisiae during serial subculturing and provide key metabolic regulation candidate targets for its long-term adaptive evolution under high-glucose stress. Full article
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19 pages, 2061 KiB  
Article
Determining the Effectiveness of Saccharomyces cerevisiae as a Postbiotic in Mass-Reared Acheta domesticus (House Cricket)
by Kimberly L. Boykin, Erik Neff and Mark A. Mitchell
Insects 2025, 16(7), 702; https://doi.org/10.3390/insects16070702 - 9 Jul 2025
Viewed by 413
Abstract
Pre-, pro-, and post-biotics have been used with success in several commercially grown insect species to increase yields and improve health outcomes. However, few studies have been published evaluating the use of nutritional supplements in Acheta domesticus. For this study, we fed [...] Read more.
Pre-, pro-, and post-biotics have been used with success in several commercially grown insect species to increase yields and improve health outcomes. However, few studies have been published evaluating the use of nutritional supplements in Acheta domesticus. For this study, we fed day-old, farm-raised, A. domesticus a Saccharomyces cerevisiae postbiotic product at one of three different inclusion rates (0, 0.25, and 0.5%) for 32 days. Crickets were analyzed for differences in average weights, total biomass, percent survival, nutritional analyses, viral qPCR, and 16S/18S microbiomes. Crickets receiving the 0.5% inclusion feed trended towards having a higher total biomass (F = 3.823, p = 0.052) and a higher percent survival per bin (F = 3.667, p = 0.057) than the crickets receiving lower inclusion feeds. No significant differences were found in viral prevalences or loads. Significant changes to the microbiomes were mostly defined by increased abundances of presumed beneficial bacteria (Akkermansia, Catenibacillus, and Odoribacter) in the groups receiving postbiotics rather than by losses of harmful bacteria. For the 18S results, there was an increased abundance of a Gregarian apicomplexan, Leidyana erratica in the treatment groups. Overall, the 0.5% inclusion feed appeared to be beneficial and further study investigating other forms of feed additives is warranted. Full article
(This article belongs to the Section Role of Insects in Human Society)
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18 pages, 2012 KiB  
Article
ATP Supply from Cytosol to Mitochondria Is an Additional Role of Aerobic Glycolysis to Prevent Programmed Cell Death by Maintenance of Mitochondrial Membrane Potential
by Akane Sawai, Takeo Taniguchi, Kohsuke Noguchi, Taisuke Seike, Nobuyuki Okahashi, Masak Takaine and Fumio Matsuda
Metabolites 2025, 15(7), 461; https://doi.org/10.3390/metabo15070461 - 7 Jul 2025
Viewed by 592
Abstract
Eukaryotic cells generate ATP primarily via oxidative and substrate-level phosphorylation. Despite the superior efficiency of oxidative phosphorylation, eukaryotic cells often use both pathways as aerobic glycolysis, even in the presence of oxygen. However, its role in cell survival remains poorly understood. Objectives: In [...] Read more.
Eukaryotic cells generate ATP primarily via oxidative and substrate-level phosphorylation. Despite the superior efficiency of oxidative phosphorylation, eukaryotic cells often use both pathways as aerobic glycolysis, even in the presence of oxygen. However, its role in cell survival remains poorly understood. Objectives: In this study, aerobic glycolysis was compared between the Warburg effect in breast cancer cells (MCF7) and the Crabtree effect in a laboratory strain of Saccharomyces cerevisiae (S288C). Methods: The metabolic adaptations of MCF7 and S288C cells were compared following treatment with electron transport chain inhibitors, including FCCP, antimycin A, and oligomycin. Results: MCF7 and S288C cells exhibited strikingly similar metabolic rewiring toward substrate-level phosphorylation upon inhibitor treatment, suggesting that mitochondrial oxidative phosphorylation and cytosolic substrate-level phosphorylation communicate through a common mechanism. Measurement of mitochondrial membrane potential (MMP) and ATP concentrations further indicated that cytosolic ATP was transported into the mitochondria under conditions of reduced electron transport chain activity. This ATP was likely utilized in the reverse mode of H+/ATPase to maintain MMP, which contributed to the avoidance of programmed cell death. Conclusions: These results suggest that the ATP supply to mitochondria plays a conserved role in aerobic glycolysis in yeast and mammalian cancer cells. This mechanism likely contributes to cell survival under conditions of fluctuating oxygen availability. Full article
(This article belongs to the Section Cell Metabolism)
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19 pages, 8164 KiB  
Article
A Novel PCR-Based Tool to Trace Oenological Saccharomyces cerevisiae Yeast by Monitoring Strain-Specific Nucleotide Polymorphisms
by Anna Baldisseri, Davide Santinello, Sara Granuzzo, Martina Frizzarin, Fabio De Pascale, Geppo Sartori, Paolo Antoniali, Stefano Campanaro and Raffaele Lopreiato
Foods 2025, 14(13), 2379; https://doi.org/10.3390/foods14132379 - 4 Jul 2025
Viewed by 337
Abstract
Saccharomyces cerevisiae plays a fundamental role in winemaking, not only driving alcoholic fermentation but also producing secondary metabolites that contribute to the organoleptic properties of wine. To ensure consistent quality and process efficiency, wineries commonly employ selected starter strains. Accordingly, the ability to [...] Read more.
Saccharomyces cerevisiae plays a fundamental role in winemaking, not only driving alcoholic fermentation but also producing secondary metabolites that contribute to the organoleptic properties of wine. To ensure consistent quality and process efficiency, wineries commonly employ selected starter strains. Accordingly, the ability to control strain purity and traceability is of critical importance. Currently, the inter-delta PCR method is widely used for the strain-specific genotyping of S. cerevisiae. However, its resolution diminishes when analyzing genetically similar strains, such as those isolated from related grape types or during genotyping of large yeast collections. To address this limitation, we developed a novel strategy that integrates computational and experimental approaches to identify highly specific allelic variants (single nucleotide polymorphisms, SNPs) within the S. cerevisiae genome. Comparative genomic analysis of twenty-eight different strains led to the identification of multiple strain-specific SNPs. From these, nine SNPs spanning five strains were selected and validated through targeted PCR assays. These assays confirmed the feasibility of using SNPs as reliable genetic markers for strain discrimination and traceability. Overall, our findings demonstrate that this SNP-based approach, implemented via multiplex allele-specific (AS) PCR assays, offers a rapid, cost-effective, and highly discriminatory alternative to current genotyping methods, particularly for differentiating closely related strains. Full article
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