Search Results (313)

Objectives: Saccharomyces boulardii CNCM I-745, a probiotic yeast, is effectively used for the treatment of acute diarrhea as well as for the prevention and treatment of traveller‘s diarrhea and diarrhea under tube feeding. The underlying mechanisms are not fully elucidated. Both antitoxic and regulatory effects on the intestinal barrier, mediated either by the yeast or yeast-derived substrates, have been discussed. Methods: To examine the effects of Saccharomyces boulardii released substrates (S.b.S) on gastrointestinal (GI) barrier function, a murine small intestinal organoid cell model under stress was used. Stress was induced by lipopolysaccharide (LPS) exposure or withdrawal of growth factors from cell culture medium (GF_Red). Stressed organoids were treated with S.b.S (200 µg/mL), and markers of GI barrier and inflammatory response were assessed. Results: GF_Red-induced stress was characterized by disturbances in selected tight junction (TJ) (p < 0.05), adherent junction (AJ) (p < 0.001), and mucin (Muc) formation (p < 0.01), measured by gene expressions, whereby additional S.b.S treatment was found to reverse these effects by increasing Muc2 (from 0.22 to 0.97-fold change, p < 0.05), Occludin (Ocln) (from 0.37 to 3.5-fold change, p < 0.0001), and Claudin (Cldn)7 expression (from 0.13 ± 0.066-fold change, p < 0.05) and by decreasing Muc1, Cldn2, Cldn5, and junctional adhesion molecule A (JAM-A) expression (all p < 0.01). Further, S.b.S normalized expression of nucleotide binding oligomerization domain (Nod)2- (from 44.5 to 0.51, p < 0.0001) and matrix metalloproteinase (Mmp)7-dependent activation (from 28.3 to 0.02875 ± 0.0044 ** p < 0.01) of antimicrobial peptide defense and reduced the expression of several inflammatory markers, such as myeloid differentiation primary response 88 (Myd88) (p < 0.01), tumor necrosis factor α (Tnfα) (p < 0.01), interleukin (IL)-6 (p < 0.01), and IL-1β (p < 0.001). Conclusions: Our data provide new insights into the molecular mechanisms by which Saccharomyces boulardii CNCM I-745-derived secretome attenuates inflammatory responses and restores GI barrier function in small intestinal organoids. Full article

Calcium microcapsules were developed by spray-drying using mannoproteins (MPs) extracted from brewer’s spent yeast, xanthan gum (XG), and maltodextrin as encapsulating materials. The formulas included 11 g of calcium, 24 g of MP, and 0, 2, 4, or 8 g of XG 100 g⁻¹ solids, obtaining C1, C2, C3, and C4 microcapsules, respectively. Maltodextrin was added to complete 100 g of solids. Calcium intestinal (IB), colonic (CB), and total bioaccessibility (TB) were estimated after a simulated gastrointestinal digestion followed by in vitro colonic fermentation. The macromolecules of microcapsules interacted by ionic and hydrophobic forces. Microcapsules C1 and C2 showed a spherical shape. However, the addition of XG to the formulation contributed to the formation of concavities in the microcapsules. All microcapsules had higher IB than the control (CaCl₂), probably due to the calcium-chelating peptides dialyzed from MP. Moreover, C1 and C2 showed the highest IB values (≈23%). However, C3 and C4 showed the highest CB values (≈11%), attributing this effect to the short-chain fatty acids produced during colonic fermentation. Finally, C1 and C2 showed the highest TB (31.8 ± 0.1 and 32.0 ± 0.4%, respectively). The use of MP allowed for obtaining a supplement with high calcium bioaccessibility. Full article

Chloroplasts, as the organelles primarily responsible for photosynthesis, require a substantial supply of iron ions. Conversely, due to Fe toxicity, the homeostasis of these ions is subject to tight regulation. Permease in chloroplast 1 (PIC1) has been identified as the primary iron importer into chloroplasts. However, previous studies suggested the existence of a distinct pathway for Fe transfer to chloroplasts, likely involving mitoferrin-like 1 (MFL1) protein. In this work, Arabidopsis MFL1 (AtMFL1) and its cucumber homolog (CsMFL1) were characterized using, among others, Arabidopsis protoplasts as well as both yeast and Arabidopsis mutants. Localization of both proteins in chloroplasts has been shown to be mediated via an N-terminal transit peptide. At the gene level, MFL1 expression profiles differed between the model plant and the crop plant under varying Fe availability. The expression of other genes involved in chloroplast Fe homeostasis, including iron acquisition, trafficking, and storage, was affected to some extent in both AtMFL1 knockout and overexpressing plants. Moreover, root growth and photosynthetic parameters changed unfavorably in the mutant lines. The obtained results imply that AtMFL1 and CsMFL1, as putative chloroplast iron transporters, play a role in both iron management and the proper functioning of the plant. Full article

Peptides play a crucial role in the development of pharmaceuticals and functional foods. Multiple studies have shown that natural bioactive peptides possess antioxidant, antihypertensive, anti-tumor, and anti-inflammatory activities. Marine bioactive peptides, especially those sourced from fish, constitute a substantial reservoir of these molecules. Although considerable research has been undertaken on fish-derived peptides, studies specifically concerning those from tuna are limited. Tuna, a marine fish of high nutritional value, generates substantial by-product waste during fishing and processing. Therefore, it is essential to conduct an evaluation of the advancements in study on tuna-derived active peptides and to offer a perspective on the direction of future investigations. This review integrates prospective bioactive peptides derived from tuna and reports contemporary strategies for their investigation, including extraction, purification, screening, identification, and activity evaluation procedures, including Yeast Surface Display (YSD) and molecular docking. This review seeks to promote the continued investigation and application of bioactive peptides derived from tuna. Full article

The number of fungal infections is steadily increasing, with considerable morbidity and mortality. Additionally, antifungal resistance is a growing concern, highlighting the need to develop new treatment options. One alternative is the use of antimicrobial peptides (AMPs). The aim of this study was to assess the in vitro and in vivo antifungal activity of designed short AMPs, Act-6 and Act 8-20, derived from cecropin transcripts of beetles from the family Scarabaeidae, against eight reference strains of the pathogenic yeasts Candida and Cryptococcus. We also evaluated the effect of these modified AMPs on the biofilm, morphogenesis, and cell morphology of Candida albicans, as well as the in vivo activity via a murine model of disseminated candidiasis. The AMPs herein analyzed exhibit differential antifungal activity against the yeasts assessed, and inhibit biofilm, hyphae, and pseudohyphae formation with morphological alterations in C. albicans. Moreover, the fungal load in mice treated with these AMPs significantly decreased. Altogether, our results suggest that Act-6 and Act 8-20 are promising antifungal molecules to control mycoses. Full article

Background/Objectives: The potential of DNA as an information-dense storage medium has inspired a broad spectrum of creative systems. In particular, hybrid biomolecular systems that integrate new materials and chemistries with DNA could drive novel functions. In this work, we explore the potential for proteins to serve as molecular file addresses. We stored DNA-encoded data in yeast and leveraged yeast surface display to readily produce the protein addresses and make them easy to access on the cell surface. Methods: We generated yeast populations that each displayed a distinct protein on their cell surfaces. These proteins included binding partners for cognate antibodies as well as chromatin-associated proteins that bind post-translationally modified histone peptides. For each specific yeast population, we transformed a library of hundreds of DNA sequences collectively encoding a specific image file. Results: We first demonstrated that the yeast retained file-encoded DNA through multiple cell divisions without a noticeable skew in their distribution or a loss in file integrity. Second, we showed that the physical act of sorting yeast displaying a specific file address was able to recover the desired data without a loss in file fidelity. Finally, we showed that analog addresses can be achieved by using addresses that have overlapping binding specificities for target peptides. Conclusions: These results motivate further exploration into the advantages proteins may confer in molecular information storage. Full article

In this study, camel milk (CM) and Gir cow milk (GCM) were fermented through cofermentation via yeast–lactic cultures, i.e., Lacticaseibacillus rhamnosus (M9, MTCC 25516) and Saccharomyces cerevisiae (WBS2A, MG101828), and their antioxidant and antidiabetic effectiveness were studied. To optimize the growth conditions, the level of proteolysis was evaluated by exploring various inoculation levels (1.5, 2.0 and 2.5%) as well as incubation durations (0, 12, 24, 36 and 48 h). Peptides were extracted and purified through 2D gel electrophoresis as well as SDS–PAGE. Water-soluble extracts (WSEs) of ultrafiltered (UF) peptide fractions were evaluated via reversed-phase high-performance liquid chromatography (RP-HPLC) to identify the peptide segments. By applying the Peakview tool, peptide sequences obtained from liquid chromatography–mass spectrometry (LC/MS) were reviewed by comparison with those in the BIOPEP database. Furthermore, the elevated levels of TNF-α, IL-6, IL-1β and nitric oxide (NO) in RAW 267.4 cells treated with lipopolysaccharide (LPS) are considerably lower than those in cultured CM and GCM. Protein macromolecules in CMs and GCMs have been captured via confocal laser scanning microscopy (CLSM) and Fourier transform infrared (FTIR) spectroscopy both before and after fermentation. Full article

The growing interest in health-promoting diets has brought fermented foods into the spotlight due to their unique microbial compositions and bioactive metabolites. Fermented foods and their beneficial microbiota are expected to stimulate the overall industry’s expansion over the next few years as their beneficial health effects become established. This narrative review explores the evolving dynamics of fermented food microbiota and their interactions with the gut microenvironment, emphasizing strategic pathways to enhance human health. Fermented foods, both industrially produced and traditionally prepared, serve as carriers of beneficial microorganisms such as lactic acid bacteria, yeasts, and certain fungi that transform food substrates into bioactive compounds including short-chain fatty acids (SCFAs), exopolysaccharides, and bioactive peptides. Simultaneously, their bioactive metabolites are the subject of passionate investigation by the scientific community, uncovering novel beneficial aspects that have not been elucidated until now. These metabolites contribute to improved gut barrier function, modulation of immune responses, and overall metabolic health. Notably, microbial fermentation can reshape the intrinsic properties of food, offering therapeutic potential beyond basic nutrition. The interactions between food-derived microbes and the host gut microbiota suggest a synergistic mechanism influencing gastrointestinal and systemic health outcomes. Nevertheless, there remains a significant gap in the comprehensive evaluation of the existing literature in this specific research area. Further research is needed to standardize fermented food formulations, validate the effects of individual microbial strains, and optimize their application in personalized nutrition and functional food development. Accordingly, this review highlights the association between the microbiota of fermented foods and their metabolites with the gut microenvironment, emphasizing their potential health-promoting properties. Full article

Introduction: Candidozyma auris (Cz. auris) has emerged globally, and diseases caused by it are associated with a mortality rate of 30–72%. This yeast is often multidrug-resistant and challenging to treat. A synthetic peptide, consisting of 11 amino acids of human lactoferrin (hLF1-11), offers a new therapy that is active against Candida albicans, non-albicans Candida yeasts, as well as Cz. auris. The current study examined the susceptibility of clinically relevant Candida species to hLF(1-11) in vitro and investigated the synergistic interaction of this peptide with fluconazole (FLU) and anidulafungin (ANI). Methods: Susceptibility of the yeasts to hLF(1-11) was tested with a microdilution method to determine minimum inhibitory concentrations (MICs). A total of 59 strains belonging to 16 species of Candida or Candidozyma were tested. The treatment cohort included 20 strains of Cz. auris originating from six different countries. Results: Mean MIC values of all susceptible strains ranged from 16.66 ± 6.46 μg/mL to 45.83 ± 10.21 μg/mL. There were no statistical differences in the susceptibility of hLF(1-11) for Cz. auris across geographic origins. In the combinatory tests, drugs acting together, the fractional inhibitory concentration indexes [FIC] < 1.0, showed a synergistic or additive effect on the efficacy of FLU and ANI when used in combination with hLF(1-11). [FIC] indexes 1–2 were interpreted as intermediate. MIC values in combinatory use were 1–2 titer steps lower than when used alone. Conclusions: hLF(1-11) inhibits the growth of yeasts that belong to the genus Candida, including Cz. auris. The combinatory use may be further investigated to treat infections caused by resistant yeasts. Full article

Penicillium expansum, a major postharvest pathogen, causes blue mold decay in apples, resulting in substantial economic losses and mycotoxin contamination. Despite the importance of effector proteins in fungal pathogenicity, the role of metalloproteases in P. expansum remains unclear. Here, we characterize an effector candidate, PeMep, through whole genome sequencing and functional analyses. Functional validation confirmed the secretory capacity of its signal peptide via yeast assays and subcellular localization. Deletion of PeMep significantly impaired fungal growth (23% reduction), conidiation (23.3% decrease), and germination efficiency. The ΔPeMep mutant exhibited hypersensitivity to osmotic, oxidative, and thermal stresses, highlighting its vital role in environmental adaptability. Importantly, pathogenicity assays revealed attenuated virulence in the ΔPeMep mutant, with 15–30% smaller lesion sizes on apples and delayed hyphal penetration compared to the wild-type, demonstrating that PeMep is essential for the pathogenic process of P. expansum 3.3703. These findings identify PeMep as a potential multifunctional effector protein crucial for fungal development, environmental adaptation, and pathogenicity in P. expansum 3.3703, providing a novel target for postharvest disease management. Full article

The World Health Organization (WHO) prioritized 19 fungal species based on the significant impact of these pathogens on human health, including the emergence of antifungal resistance, which highlights the necessity of finding new antifungal therapies. Among these novel therapeutic approaches, the antimicrobial pore-forming peptide C14R has shown to be promising against Candida albicans and Candida auris. In this study, the antifungal in vitro efficacy of C14R was assessed against six additional species from the WHO priority list, Cryptococcus neoformans, Cryptococcus gattii, Candida glabrata, Candida tropicalis, Candida parapsilosis and Candida krusei, as well as against Candida dubliniensis. This study shows that C14R has good antifungal activity against several clinical isolates of the studied species, with MIC values between 0.8476 and 10.88 µg/mL. Most notably, some of the studied isolates are resistant to commonly used antifungal drugs but are susceptible to the peptide. C14R showed, moreover, its capacity to disrupt Cryptococcus capsules, beyond its already proven capacity to disrupt plasma membranes, and its antifungal activity was not affected depending on the serotype or species assessed. The inclusion of basidiomycete and ascomycete yeasts allowed us to display the broad-spectrum potential of C14R, highlighting it as a promising candidate as an antifungal agent. Full article

Surface display technology has revolutionized whole-cell biocatalysis by enabling efficient enzyme immobilization on microbial cell surfaces. Compared with traditional enzyme immobilization, this technology has the advantages of high enzyme activity, mild process, simple operation and low cost, which thus has been widely studied and applied in various fields. This review explores the principles, optimization strategies, applications in the food industry, and future prospects. We summarize the membrane and anchor protein structures of common host cells (Escherichia coli, Bacillus subtilis, and yeast) and discuss cutting-edge optimization approaches, including host strain genetic engineering, rational design of anchor proteins, innovative linker peptide engineering, and precise regulation of signal peptides and promoters, to maximize surface display efficiency. Additionally, we also explore its diverse applications in food processing and manufacturing, additive synthesis, food safety, and other food-related industries (such as animal feed and PET packaging degradation), demonstrating their potential to address key challenges in the food industry. This work bridges fundamental research and industrial applications, offering valuable insights for advancing agricultural and food chemistry. Full article

Cottonseed is a valuable source of high-quality proteins and oils. Defatted cottonseed meal (DCSM), a by-product of cottonseed oil extraction, holds significant potential as a sustainable protein resource. This review outlines the chemical composition, structural features, and unique properties of cottonseed, with a focus on its inherent antinutritional factors, such as gossypol. Strategies for enhancing the utilization of DCSM as a protein source are systematically evaluated, including physical, chemical, and biological methods used to eliminate or reduce antinutritional components. Among these, microbial fermentation, particularly solid-state fermentation, is highlighted as a promising, eco-friendly approach for detoxification and nutritional improvement. This review further discusses critical factors influencing the removal of anti-nutritional compounds, such as pretreatment methods, fermentation parameters, and microbial strains. The efficacy of probiotic strains (e.g., Bacillus and yeasts) in enhancing the protein digestibility, amino acid profiles, and functional properties of DCSM is discussed. Additionally, recent advances in the application of fermented cottonseed protein in foods (e.g., animal feed, functional peptides, and food additives) and non-food sectors (e.g., biofuels and bioplastic) are explored. The integration of probiotic-driven fermentation processes is proposed as a strategy to exploit the full nutritional and economic potential of DCSM, paving the way for its broader and sustainable use in foods and non-food applications. Full article

The Rare Cold-Inducible 2 (RCI2) gene encodes a conserved hydrophobic peptide that plays a crucial role in ion homeostasis, membrane stability, and responses to abiotic stress. In this study, six members of the MsRCI2 gene family were identified in Medicago sativa L., all of which contain highly conserved PMP₃ domains. Comparative collinearity analysis revealed syntenic relationships between M. sativa and M. truncatula, with each gene displaying distinct expression profiles under various stress conditions. Among them, MsRCI2B was significantly upregulated in response to salt stress. Alfalfa plants overexpressing MsRCI2B exhibited enhanced salt tolerance, as evidenced by increased antioxidant enzyme activities and reduced accumulation of malondialdehyde (MDA), hydrogen peroxide (H₂O₂), and superoxide anion (O₂⁻) compared to wild-type plants. Furthermore, the transgenic lines maintained better Na⁺/K⁺ homeostasis under salt stress, reflected by a lower Na⁺/K⁺ ratio and significantly elevated expression of key ion transport genes, including MsSOS1, MsAKT1, and MsNHX1. To elucidate the molecular mechanisms underlying MsRCI2B function, a yeast two-hybrid (Y2H) screen identified 151 potential interacting proteins. Gene Ontology (GO) enrichment analysis revealed that these interactors are mainly involved in antioxidant defense and ion transport. Further validation confirmed direct interactions between MsRCI2B and both calmodulin (CaM) and vacuola H⁺-ATPase (V-H⁺-ATPase), suggesting that MsRCI2B contributes to ion homeostasis through interactions with CaM and V-H⁺-ATPase, thereby promoting Na⁺/K⁺ balance and enhancing salt tolerance. These findings provide new insights into the role of MsRCI2B in salt stress responses and underscore its potential as a genetic target for enhancing salinity tolerance in forage crops. Full article

The objective of this study was to determine whether hydroxy-selenomethionine (OH-SeMet) exerts better protective effects on sows against heat stress than sodium selenite (SeNa) or seleno-yeast (SeY). A total of 60 sows (Landrace × Yorkshire) were randomly allocated into the three groups and fed a base diet supplemented with SeNa, SeY, or OH-SeMet at 0.3 mg Se/kg under a heat stress condition for a reproductive cycle. Compared to SeNa or SeY, OH-SeMet could more effectively sustain offspring growth performance, as evidenced by an increased number of live-born piglets, higher litter weight at day 21, and greater litter body weight gain from days 1 to 21. OH-SeMet was more effective in supporting endogenous redox systems, as shown by enhanced levels of TXNRD and GSH and reduced levels of GSSG in the serum of sows, improved T-AOC, TXNRD, and GSH alongside decreased MDA and GSSG in the serum of piglets, and heightened T-AOC in the jejunum of piglets. Furthermore, among the two tested organic Se sources, OH-SeMet was more effective than SeY in regulating immune responses compared to SeNa. OH-SeMet reduced inflammation-related markers CRP, HP, MAP, LPS, IL-1β, IL-6, and TNF-α, some or all of which were reduced in the serum of sows and their offspring. In addition, OH-SeMet also showed reduced glucose, TG, and NEFA levels, along with elevated insulin levels in the serum of sows. Correspondingly, among the two organic forms of Se, particularly those sows fed OH-SeMet showed better gut protection for the sows’ offspring, as indicated by a reduced crypt depth and increased villus height/crypt depth ratio in the duodenum, jejunum, and ileum than those fed SeNa. Specifically, compared to SeNa or SeY, OH-SeMet upregulated the expression of selenoproteins (GPX6, TXNRD3, GPX4, and SELENON), the tight junction protein (ZO-1), and host defense peptide gene (pBD1, pBD2, pBD3, NPG3, NPG4), along with downregulating levels of inflammation factor (IL-1β, IL-6 and TNF-α) and pro-apoptotic factor (P53) in the jejunum of piglets. Taken together, OH-SeMet more effectively mitigated the adverse effects induced by heat stress in sows and their offspring. Full article