Search Results (337)

Pulses are recognized as sustainable foods due to their high nutritional density, low environmental footprint, and versatility as plant-based ingredients. Fermentation has emerged as a powerful bioprocessing tool to further enhance nutritional, sensory, techno-functional, and health-promoting properties of pulses. This review summarizes recent advances in the fermentation of commonly consumed pulses using lactic acid bacteria, yeasts, molds, and co-fermentation microorganism consortia, focusing on the biochemical mechanisms underlying changes in their nutritional and bioactive potential. Microbial metabolism (i.e., α-galactosidase and phytase activity) reduces antinutritional factors, such as raffinose family oligosaccharides and phytic acid, while promoting the release of bound nutrients and bioactive compounds as phenolics, increasing their bioaccessibility and bioactivity. Microbial amylases change the carbohydrate profile by decreasing simple sugars, modifying starch digestibility, and favoring resistant starch production. Microbial lipases remodel lipids, improving the fatty-acid distribution and nutritional value. Protein hydrolysis by microbial proteases enhances digestibility and generates bioactive peptides with antioxidant and antihypertensive properties, among others. Co-fermentation systems offer additional opportunities to tailor metabolic outcomes, facilitating positive symbiotic interactions between microorganisms. Overall, fermentation represents a key technology to unlock the full potential of pulses as next-generation ingredients, supporting the development of nutritious, functional, and sustainable foods for future food systems. Full article

Serine proteases represent a significant family of proteolytic enzymes, characterized by their serine-dependent catalytic mechanism. These enzymes are integral to various biological processes, including fungal growth, development, and pathogenicity. Despite their importance, the sequence characterization and expression patterns of this protein family in Setosphaeria turcica are not yet fully characterized and remain underexplored. A total of 74 putative serine protease family proteins (StSPs) were identified in S. turcica and classified into 12 subfamilies based on phylogenetic analysis. Structural domain analysis revealed that 24 StSPs contain signal peptides, of which five were experimentally validated as secretory proteins through yeast secretion assays. Expression profiling using RNA-seq data demonstrated that StSPs exhibit distinct expression patterns across different developmental and infection stages, with 61 genes showing high expression during critical infection phases. The expression levels of nine genes were validated via qRT-PCR, and the results were consistent with the RNA-seq data. Among these proteins, StSP8-4 demonstrated elevated expression during the course of fungal infection. Functional characterization of StSP8-4 OE and RNAi strains revealed that this gene plays a crucial role in maintaining fungal pathogenicity, although silencing did not impair conidium or hyphal development. These findings provide valuable insights for further research on serine protease genes in S. turcica. Full article

Floral volatile terpenoids are known to play important roles in plant pollination biology by attracting animal pollinators, repelling antagonists, and enhancing resistance to potential microbial pathogens. The terpenoid blend emitted by a flower is usually plant-lineage specific and is primarily determined by a set of versatile terpene synthases (TPSs), which catalyze the final step of diverse terpenoid synthesis. The strongly scented flower of Jasminum sambac (L.) Aiton emits linalool and α-farnesene, which dominate the nocturnal floral VOCs, yet the corresponding TPSs have not been identified. Here, we show that four TPS enzymes are responsible for the synthesis of a mixture of volatile terpenoids in the flower, based on their highly correlated and almost exclusive expression in the petal, as well as their enzymatic characterizations in vitro and in Nicotiana benthamiana Domin. JsTPS01 (TPS-a) acts as a sesquiterpene synthase, producing τ-cadinol in yeast at levels that mirror its rhythmic expression in petals. JsTPS02 (TPS-b) carries a plastid-targeting transit peptide, localizes to chloroplasts/plastids, and converts geranyl diphosphate (GPP) to linalool with high affinity (K_m = 28.2 ± 3.4 µM). JsTPS03 is a TPS-b clade member that can convert farnesyl diphosphate (FPP) to farnesol with a K_m of 14.4 ± 5.9 μM in an in vitro assay using isolated yeast vehicles. JsTPS04 (TPS-e/f) exhibits dual targeting—cytosolic in protoplasts of Arabidopsis thaliana (L.) Heynh, but plastidic in J. sambac petals—and functions as a bifunctional mono-/sesqui-TPS, forming linalool from GPP (K_m = 2.5 ± 0.3 µM) and trans-nerolidol from FPP (K_m = 7.6 ± 0.6 µM). Transient expression in N. benthamiana leaves further confirmed its in-planta linalool production. Collectively, we identified four preferentially expressed terpene synthases that contribute to the production of linalool, τ-cadinol, trans-nerolidol, and farnesol in J. sambac. Full article

γ-Aminobutyric acid (GABA) is a bioactive metabolite valued in functional foods, but its microbial production is strongly influenced by nutrient availability. Levilactobacillus brevis CRL 2013 is an efficient GABA producer; however, its biosynthesis depends on culture medium composition. In this study, integrated physiological, proteomic, and transcriptional analyses were applied to assess the influence of nitrogen source composition and concentration on GABA production. No extracellular GABA was detected in a chemically defined medium containing all amino acids and glutamate (CDMg), whereas supplementation with yeast extract or Casitone restored high-level production. The highest GABA accumulation (~250 mM) was obtained in CDMg supplemented with 1% yeast extract or 2% Casitone, and a clear peptide dose-dependent effect was observed. In contrast, other protein hydrolysates or free amino acids alone did not stimulate GABA synthesis. Proteomic analysis revealed overexpression of the key enzyme GadB and changes in nucleotide and fatty acid pathways. Transcriptional analysis confirmed that peptide supplementation was accompanied by increased transcription of the gadRCB–gltX operon, in agreement with GABA accumulation. Overall, these results demonstrated that peptide composition and availability are critical determinants of GABA biosynthesis in Lv. brevis CRL 2013and provide a basis for optimizing peptide-based media to enhance GABA formation in food fermentations. Full article

Parasitoid wasp venoms represent highly specialized biochemical arsenals that evolved to manipulate host physiology and ensure successful development of the parasitoid offspring. However, the molecular targets and mechanisms underlying this complex host modulation remain poorly understood. To address this, we employed an AI-driven discovery pipeline, integrating the sequence-based predictor D-SCRIPT with the structural modeler AlphaFold3, to characterize LjSPI-1, a venom serpin from Liragathis javana. This computational workflow highlighted a previously unreported candidate partner—a host serine carboxypeptidase (Chr09G02510). Crucially, we detected a direct physical interaction between these two proteins through both in vitro pull-down and in vivo yeast two-hybrid assays, supporting this AI-prioritized interaction under experimental conditions. Our study identifies a high-priority molecular pairing and demonstrates the utility of an AI-guided strategy for uncovering candidate targets of venom proteins. In addition, guided by the predicted biochemical role of Chr09G02510, we propose several plausible physiological hypotheses linking this interaction to host peptide metabolism and immune modulation. These hypotheses serve as a conceptual basis for future mechanistic and toxicological investigations. Full article

Fermented mare’s milk, or koumiss, has been consumed for centuries across Central Asia for its nutritional and therapeutic value. Mare’s milk differs from bovine milk by its near 1:1 casein-to-whey ratio, high lysozyme and lactoferrin, abundant immunoglobulins, and low β-lactoglobulin, which enhance digestibility, reduce allergenicity, and increase antimicrobial activity. During fermentation, lactic acid bacteria and yeasts transform this substrate into a reservoir of bioactive proteins, peptides, and metabolites. Multi-omics profiling has identified more than 2300 peptides and over 350 metabolites, including sequences with angiotensin-converting enzyme (ACE)-inhibitory, antioxidant, antimicrobial, and immunomodulatory activities. Preclinical and limited clinical data indicate potential benefits for lipid metabolism, cardiovascular function, and gut health. Mechanistically, these effects appear to arise from synergistic actions of native proteins, fermentation-derived peptides, and probiotic consortia. Technological advances such as rational starter culture design, controlled proteolysis, and microencapsulation offer strategies to enhance bioactive yield and stability. However, standardized fermentation protocols and clinical validation remain necessary to position koumiss as a scientifically supported functional food. Full article

This study evaluated the nutritional, functional, biological, and sensory potential of proteins derived from Yarrowia lipolytica biomass and their enzymatic hydrolysates for food applications. Three strains were cultivated under bioreactor conditions, with strain JII1c selected for its superior biomass yield and protein content. Its amino acid composition was rich in lysine and branched-chain amino acids, with protein quality indices (CS = 37.8%, EAAI = 36.17%) confirming value in plant-based diets. Proteins were isolated and hydrolysed using a non-commercial serine protease from Cucurbita ficifolia, which enhanced solubility (NSI: 19.4 → 49.2%), water and oil absorption, and emulsion stability. Hydrolysates showed notable biological activities, including ACE (71.8%), DPP-IV (52.3%), and α-glucosidase (67.4%) inhibition, indicating potential metabolic benefits. Sensory evaluation of extrudates confirmed improvements in aroma, texture, and flavour when hydrolysates were incorporated. The use of a plant-derived protease demonstrates a sustainable approach to producing bioactive peptides. Y. lipolytica hydrolysates emerge as promising clean-label ingredients that combine nutritional quality with techno-functional performance, supporting their integration into health-oriented and sustainable food products. Full article

Bacteria are promising sources of bioflocculants, yet their regulatory machinery for bioflocculant synthesis remains underexplored. This study focused on evaluating the biosynthetic genes, optimisation and assessment of bioflocculation efficiency in wastewater. The isolated bioflocculant producers were identified by 16S rRNA and rpoB gene analysis. Polymerase chain reaction was used to assess the presence of polyketide synthase I (PKS-1), polyketide synthase II (PKS-II), non-ribosomal peptide synthetase (NRPS), epsH and epsJ. A one-factor-at-a-time technique was utilised for optimisation of culture conditions. The bioflocculants’ efficiencies were evaluated in wastewater using the Jar test method. Among 31 isolates, Klebsiella michiganensis and Klebsiella pasteurii were the most potent bioflocculant producers. They both revealed the presence of PKS-II. K. pasteurii possessed the epsH gene. The optimal conditions for maximum bioflocculant production (95% activity) by K. michiganensis were a temperature of 35 °C, pH of 5, galactose, tryptophan and 84 h of incubation. K. pasteurii’s maximum bioflocculant production of 83% was obtained at a temperature of 35 °C and pH of 7, with galactose, a mixture of urea, yeast extract, and ammonium sulphate (NH₄)₂SO₄ and 96 h of fermentation. Their bioflocculants reduced the chemical oxygen demand and turbidity of wastewater by more than 70%. The bacteria had promising bioflocculant production with potential applicability in wastewater treatment. Full article

Background/Objectives: Vaccine elicitation of antibodies with high HIV-1 neutralization breadth is a long-standing goal. Recently, the induction of such antibodies has been achieved at the fusion peptide site of vulnerability. Questions remain, however, as to how much anti-fusion peptide antibodies can be improved and whether their neutralization breadth and potency are sufficient to prevent HIV-1 infection. Methods: Here, we use yeast display coupled with deep mutational screening and biochemical and structural analyses to study the improvement of the best fusion peptide-directed, vaccine-elicited antibody, DFPH_a.01, with an initial 59% breadth. Results: Yeast display identified both single and double mutations that improved recognition of HIV-1 envelope trimers. We characterized two paratope-distal light chain (LC) mutations, S10R and S59P, which together increased breadth to 63%. Biochemical analysis demonstrated DFPH-a.01_10R59P-LC, and its component mutations, to have increased affinity and stability. Cryo-EM structural analysis revealed elbow-angle influencing by S10R-LC and isosteric positioning by S59P-LC as explanations for enhanced breadth, affinity, and stability. Conclusions: These results, along with another antibody with enhanced performance (DFPH-a.01_1G10A56K-LC with 64% breadth), suggest that mutations improving DFPH_a.01 are plentiful, an important vaccine insight. Full article

Background: Whether therapeutic albumin (ThHSA) can serve as a defense tool in Candida species (spp.) infections is still a matter of debate, although many physicians are in the habit of infusing ThHSA to restore the physiological concentration of endogenous human serum albumin (HSA). Given the need for innovative anti-Candida strategies, we assessed in vitro the role of ThHSA alone or in combination with voriconazole (VCZ) in combating Candida spp. growth and the role of bovine serum albumin (BSA)—used as a substitute for HSA—with two endogenous bovine antimicrobial peptides in combating C. albicans and other microbes. Results: The combination of ThHSA with VCZ enhanced the antifungal effect on C. albicans, sensitive C. tropicalis, sensitive C. glabrata, and C. lusitaniae. However, for resistant C. tropicalis, the combination of ThHSA with VCZ promoted yeast growth, and VCZ tended to suppress the antimicrobial effect of ThHSA on resistant C. glabrata. As to the possible transposition of ThHSA-type properties to BSA (as regards the growth inhibition of other pathogens), we tested combinations of BSA with two physiological chromogranin A-derived antimicrobial peptides (catestatin and cateslytin). BSA enhanced significantly the activity of catestatin (but not cateslytin) in combating C. albicans, A. fumigatus, and M. luteus, but was inactive against S. aureus and E. coli. Conclusions: Our experiments support the fact that albumins display intrinsic antimicrobial properties, with an unpredictable growth inhibitory effect on various microbes. ThHSA can thus be an adjunctive tool for more efficient care of some, though not all, infections. The interaction of BSA with catestatin and cateslytin is related to their structure, with BSA significantly enhancing the effect of catestatin but not that of cateslytin. Full article

The incorporation of probiotic microorganisms into fermented foods has long been recognized as a promising strategy to enhance gut health and overall well-being. Conventional probiotics, mainly from the bacterial genera Lactobacillus, Bifidobacterium, Lacticaseibacillus, Levilactobacillus, Lactiplantibacillus and yeast genus Saccharomyces, contribute to gastrointestinal homeostasis, immune modulation, and metabolic balance. Building on these foundations, recent advances in synthetic biology, systems microbiology, and genetic engineering have enabled the development of smart probiotics: engineered or selectively enhanced strains capable of sensing environmental cues and producing targeted bioactive compounds, such as neurotransmitters and anti-inflammatory peptides. These next-generation microorganisms offer precision functionality in food matrices and hold promise for applications in gastrointestinal health, immune support, and gut–brain axis modulation. However, their deployment also raises critical questions regarding biosafety, regulatory approval, and consumer acceptance. This review provides a comprehensive overview of the mechanisms of action, biotechnological strategies, and health-oriented fermentation applications of smart and functional probiotics, emphasizing their role in the future of personalized and evidence-based functional foods. Full article

Cereal grains have been dietary staples for millennia, providing essential nutrients alongside their primary carbohydrate content. Recently, the search for sustainable, nutrient-rich alternatives has drawn attention to spelt (Triticum aestivum ssp. spelta L.), a low-input crop with promising nutritional properties. Spelt supplies a higher content of unsaturated fatty acids and minerals, such as iron, zinc, and magnesium and exhibits lower levels of phytic acid compared to common wheat. This study explores the nutraceutical potential of fermented bakery products made from spelt and wheat flours using sourdough fermentation, a process driven by lactic acid bacteria (LAB) and yeasts. Breads produced with baker’s yeast were included for comparison. Specifically, this manuscript focuses on the generation of bioactive peptides (BPs), which have demonstrated anti-oxidant, anti-inflammatory, and gut-protective effects by modulating oxidative stress and inflammatory signaling pathways. By comparing aqueous extracts from breads prepared with varying flours and fermentation methods, optimal conditions for producing functional baked goods could be defined. The findings may offer new avenues for developing bakery products that potentially increase intestinal health while promoting sustainable agriculture through the use of spelt. Full article

Inflammatory facial dermatoses (atopic dermatitis [AD], acne vulgaris, contact dermatitis, seborrheic dermatitis, rosacea, perioral dermatitis, and demodicosis, etc.) often profoundly impact patients’ appearance and psychological well-being. In this narrative review, we wanted to present the current knowledge on the role of skin microbiota in common facial dermatoses. Skin keratinocytes are the primary producers of antimicrobial peptides (AMPs) and express Toll-like receptors (TLRs), which stimulate the T helper (Th1) immune response, with the production of interferon (IFN). They can also produce certain pro-inflammatory cytokines, namely IL-1β, IL-18, IL-6, IL-10, and the tumor necrosis factor (TNF). In healthy infants, the bacterial skin microbiota is predominantly composed of Firmicutes (genera Staphylococcus and Streptococcus), as well as Actinobacteria, Proteobactera, and Bacteroidota. The genera Cutibacterium and Staphylococcus, which have antimicrobial effects and compete with pathogens for nutrients/ecological niches, coexist symbiotically on the skin and can reduce the expression of TLR2 and TLR4. In patients with AD, lesional/non-lesional skin was found to have increased colonization by Staphylococcus aureus which reduces effector T lymphocytes’ ability to produce cytokines, such as IL-17A and IFN-γ, leading to decreased AMP production and impaired skin microbiota immune functionality. In patients with rosacea, the overexpression of TLR2 may stimulate elevated pro-inflammatory cytokine production (IL-8, IL-1β, and TNF-α, etc.), exacerbating the inflammatory response. Also, increased colonization by Malassezia yeasts triggers a Th2 immune response and cytokine secretion (IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, TNF-α, beta-defensin, IFN-γ, nitric oxide, and histamine), and participates in signaling pathways. Insight into these factors may further improve clinical approaches to patients with facial dermatoses. Full article

Background/Objectives: Receptor-mediated transcytosis utilizing the native transporters at the blood–brain barrier (BBB) is a growing strategy for the delivery of therapeutics to the brain. One of the major challenges in identifying appropriate human transcytosis targets is that there is a species-specific transporter expression profile at the BBB, complicating translation of successful preclinical candidates into humans. In an effort to overcome this obstacle and identify proteins capable of binding human-relevant BBB ligands, we generated and screened a BBB-targeting library against human-induced pluripotent stem cell-derived brain microvascular endothelial-like cells (iPSC-derived BMEC-like cells). As targeting molecules, we used lamprey antibodies, known as variable lymphocyte receptors (VLRs), and generated a VLR library by immunizing lamprey with iPSC-derived BMEC-like cells, and inserting the resultant VLR repertoire into the yeast surface display system. Methods: The yeast displayed VLR library was then panned against human iPSC-derived BMEC-like cells and lead VLRs were validated using human in vitro models and mouse and human ex vivo brain tissue sections. Results: Finally, brain uptake for a set of VLRs was validated in mice. Of the 15 lead VLR candidates, 14 bound to human BBB antigens, and 10 bound to the murine BBB. Pharmacodynamic testing using the neuroactive peptide neurotensin indicated that the lead candidate, VLR2G, could cross the mouse BBB after intravenous injection and deliver sufficient neurotensin payload to generate a pharmacological response and lower systemic body temperature. Conclusions: Together, these results demonstrate the application of a novel screening technique capable of identifying a VLR with human relevance that can cross the BBB and deliver a payload. Full article

Verticillium wilt, caused by Verticillium dahliae, severely threatens various crops and trees worldwide. This study aimed to characterize the function of a CUE (coupling of ubiquitin conjugation to endoplasmic reticulum (ER) degradation)-domain-containing protein, VdCUE, in V. dahliae, which exhibits sequence divergence between the defoliating strain XJ592 and the non-defoliating strain XJ511. We generated ∆VdCUE-knockout mutants and evaluated their phenotypes in growth and virulence. Functional analyses included verifying the signal peptide activity of VdCUE, testing its ability to induce cell death or inhibit BAX-induced cell death in Nicotiana benthamiana leaves, and identifying host targets via yeast two-hybrid screening. The ∆VdCUE mutants showed reduced formation of melanized microsclerotia but no other obvious growth defects. Cotton plants infected with the ∆VdCUE mutants exhibited a significantly lower disease index and defoliation rate. VdCUE was confirmed to be secreted via a functional signal peptide, but it neither triggered cell death nor inhibited BAX-induced cell death. Three putative host targets were identified and supported by AI-based three-dimensional structural modeling, including tRNA-specific 2-thiouridylase, peptidyl-prolyl cis-trans isomerase, and 40S ribosomal protein, which may mediate VdCUE-dependent virulence regulation. These findings reveal VdCUE as a key virulence factor in V. dahliae, contributing to our understanding of its pathogenic mechanism. Full article