Search Results (2,155)

Soybean molasses, a by-product of alcohol-based soy protein concentrate production, is rich in stachyose and other functional oligosaccharides, but its high sucrose content and other fermentable non-target sugars hinder the efficient purification of stachyose. In this study, the sugar-utilization patterns of four commonly used microbial chassis or production strains, Escherichia coli W, E. coli BL21, Saccharomyces pastorianus Weihenstephan 34/70, and Komagataella phaffii (formerly Pichia pastoris) GS115, were systematically compared to identify a suitable host for selective stachyose enrichment. Among them, E. coli W showed the best performance in rapidly consuming non-target sugars while retaining stachyose. Based on this strain, a CRISPR–Cas9 engineering strategy was applied by deleting the endogenous α-galactosidase gene melA and overexpressing the sucrose permease gene cscB. The resulting strain selectively and nearly completely removed sucrose and other non-target sugars from soybean molasses, increasing the proportion of stachyose from <30% to >90% of total soluble solids. Further optimization of nitrogen source level, inoculum size, and initial °Brix improved fermentation performance. These results demonstrate an effective biological pre-purification strategy for selective stachyose enrichment from soybean molasses. Full article

Toxicological risk assessment of food ingredients has traditionally relied on identifying a no-observed-adverse-effect level (NOAEL) or benchmark dose (BMD), followed by the application of default uncertainty factors (UFs) to derive health-based guidance values (HBGVs) such as the acceptable daily intake (ADI). While effective for conventional food additives, this approach may be inappropriate for nutrients and intrinsic food components with established physiological functions. This paper critically explores these limitations using free glutamate as a central example, alongside additional cases relevant to infant nutrition, including vitamin C, iodine, and human milk oligosaccharides (HMOs). Data on free glutamate in human milk show that breastfed infants habitually ingest amounts far exceeding additive-based ADIs without adverse effects, underscoring the limitations of applying default uncertainty factors and classical toxicological paradigms to endogenous nutrients. Comparable considerations apply to protein hydrolysates and amino acid-based infant formulas evaluated by EFSA, where growth, tolerance, and compositional suitability are integral to safety assessment. Overall, nutrient safety evaluation requires an integrative, physiology-informed framework that incorporates realistic exposure, developmental stage, and metabolic competence. Breast milk provides a biologically relevant reference, supporting a proportionate and science-based application of toxicological principles in infant nutrition. Full article

Background: Paramagnetic manganese (Mn(II)) has emerged as a promising alternative to gadolinium-based contrast agents (GBCAs) due to its favorable magnetic properties. Despite extensive research, no Mn-based agent has yet achieved clinical translation. Because free Mn(II) is toxic, macromolecular complexes incorporating stable macrocyclic DOTA chelators conjugated to polysaccharides may enhance coordination stability and improve the safety profile of Mn(II)-based contrast agents. Methods: Two chemical routes, maleimide- and ester-mediated, were evaluated for covalent coupling of DOTA-based macrocyclic ligands to the backbone of selected poly- and oligosaccharides. Subsequently, DOTA-modified carboxymethyldextran, aminodextran, and chitosan oligosaccharide were labeled with paramagnetic Mn(II) under mild conditions. ATR-FTIR confirmed the successful conjugation of DOTA chelators to the sugar backbone. The conjugates were further characterized by DLS, ICP-MS, and FPLC. In vitro relaxivity was measured at high field strength to evaluate MRI performance. In vivo contrast efficacy was first assessed using in ovo MRI in chicken embryos and subsequently evaluated by biodistribution studies in nude mice. Results: In vitro relaxivity studies demonstrated higher signal enhancement of the poly-/oligosaccharide-DOTA-Mn(II) conjugates compared with MnCl₂ and the clinical agent gadoteridol (ProHance^®). In ovo MRI showed persistent vascular enhancement up to 120 min, while in nude mice, contrast enhancement was observed in the liver, kidneys, and gallbladder 40 min post-injection. Conclusions: Mn(II)-tagged sugar-based imaging probes may offer a promising non-gadolinium alternative to GBCAs, with tunable biodistribution profiles depending on carrier molecular weight. Full article

Novel ingredients from rowanberry pomace were developed for French-type bread applications via supercritical CO₂ extraction and the enzymatic and ultrasound treatment of the defatted residue (DFR), which contained 6.367% of proteins, 8.36% of soluble, and 43.04% insoluble fiber. Proteolytic enzymes from Bacillus licheniformis and Aspergillus oryzae, and cellulolytic enzyme mixtures Viscozyme L and Celuclast, were used to increase the soluble fraction. Treating DFR with enzymes generated significant amounts of soluble substances containing oligosaccharides, fructose, and glucose, with Viscozyme L being more effective than proteases. Tri-, and tetrapeptides, chlorogenic acids, and dihydroxy coumarins were also present in the soluble extracts of fermented DFR. The antioxidant characteristics of treated DFR were evaluated by the in vitro assays. Substitution of >5% of wheat flour with untreated DFR significantly reduced bread volume and crumb porosity; however, these adverse effects were mitigated by using fermented DFR. The highest bread volume (1845 cm³) and porosity (78.38%) were observed in bread containing 5% pomace that underwent enzymatic hydrolysis and ultrasound treatment. The substitution of flour with DFR significantly increased the antioxidant characteristics of bread samples and the substances generated during the in vitro digestion. It may be concluded that rowanberry pomace ingredients may improve bread nutritional quality and assist in the sustainable use of fruit processing by-products. Full article

This study focuses on Lacto-N-neotetraose (LNnT), a core component of human milk oligosaccharides. Although LNnT has been demonstrated to promote early intestinal development and maintain gut homeostasis, its protective mechanism against D-galactose-induced intestinal injury and associated cognitive impairment remains unclear. This investigation systematically examined the protective effects and underlying mechanisms of LNnT against D-gal-induced colonic damage and cognitive impairment in mice. The results demonstrated that LNnT not only significantly improved systemic physiological phenotypes and upregulated the expression of colonic tight junction proteins to repair the intestinal barrier, but also effectively enhanced learning and memory abilities in mice. Concurrently, LNnT reduced serum proinflammatory factor levels, elevated the anti-inflammatory factor IL-10, and alleviated oxidative stress. Furthermore, LNnT remodeled the gut microbiome structure by increasing microbial diversity, enhancing beneficial bacteria abundance, and promoting short-chain fatty acid production. Untargeted metabolomics analysis further revealed that LNnT corrected metabolic disturbances by regulating key sphingolipid molecules (ceramide, sphingosine, S1P) and the expression of related metabolic enzymes (ACER2, SphK2). In summary, this study suggests that LNnT mitigates intestinal injury and improves cognitive function, potentially through modulation of the gut microbiota–sphingolipid metabolism axis, although further causal validation is warranted. These findings provide a mechanistic foundation for future studies exploring its potential as a functional dietary ingredient. Full article

The significance of less abundant genera within the gut microbiota, such as Parabacteroides, remains largely unexplored. Despite its low levels, Parabacteroides is highly conserved and potentially beneficial across populations. This trial aimed to evaluate whether a four-week intake of glucolacto-oligosaccharides (GLO), previously reported as an enhancer of Parabacteroides, improves defecation frequency as the primary outcome. It also assessed holistic gut health and underlying microbiota-based mechanisms. In this randomized, double-blind, placebo-controlled trial, 50 healthy Japanese participants with a defecation frequency of five or fewer times per week were enrolled. The mean (±SE) weekly defecation frequency in the GLO group was 3.2 ± 0.2 at baseline, increasing to 5.8 ± 0.6 at week 4, whereas that in the placebo group was 3.4 ± 0.3 at baseline, increasing to 4.4 ± 0.3 at week 4. The time-dependent weekly defecation frequency was significantly higher in the GLO group than in the placebo group (p = 0.029). Changes in the relative abundance of the genus Parabacteroides significantly increased in the GLO group compared with in the placebo group. Changes in fecal bile acid composition were also confirmed in the GLO group compared with the placebo group, which was thought to be due to the unique features of Parabacteroides. Furthermore, changes in alpha diversity indices were significantly higher in the GLO group than in the placebo group (Simpson, p = 0.041; Pielou, p = 0.022). Additional analysis demonstrated that the increase in alpha diversity in the GLO group was significantly correlated with the increase in the relative abundance of Parabacteroides (p = 0.006), which tended to be associated with decreases in serum gamma-glutamyltransferase (p = 0.089) and serum triglyceride (p = 0.075) levels. These data suggest that GLO intake improved defecation status, selectively increased Parabacteroides, and harmonized the gut environment. Full article

Human milk oligosaccharides (HMOs) exhibit substantial inter-individual and secretor-dependent variation, yet comprehensive quantitative data across diverse maternal phenotypes remain limited. In this study, we analyzed 578 human milk samples from four Asian populations using a dual mass spectrometry approach, combining quadrupole time-of-flight (Q-TOF) for structural profiling and triple quadrupole (QQQ) mass spectrometry for absolute quantitation of 15 major HMOs. Samples were classified into Secretor (76.7%) and Non-Secretor (23.3%) groups based on α-1,2-fucosylated HMO profiles. Secretor milk was enriched in α-1,2-fucosylated HMOs, whereas Non-Secretor milk showed markedly reduced levels of these structures. However, Non-Secretor retained substantial total fucosylated HMOs (65–76% of Secretor levels), accompanied by increased α-1,3/4-fucosylated structures, including up to 3.2-fold higher levels of 3-fucosyllactose (3-FL). Sensitive QQQ quantitation further revealed trace levels of α-1,2-fucosylated HMOs in Non-Secretor at concentrations 10–100-fold lower than in Secretor. Correlation analysis indicated an inverse relationship between α-1,2- and α-1,3-fucosylation patterns, consistent with redistribution of fucosylation pathways. These findings suggest that the Non-Secretor phenotype represents a distinct compositional state rather than a simple loss of α-1,2-fucosylation and provide a quantitative framework for phenotype-informed nutritional strategies. Full article

Background: Intestinal barrier problems cause obesity and related health issues. We focus on treatments that fix the gut lining and change gut bacteria. Soy oligosaccharides (SOSs) are prebiotics. They change gut bacteria and help lower fats. The mechanism by which SOS affects high-fat diet (HFD)-induced obesity remains to be fully elucidated. Objectives: We want to see if SOS improves the mucus barrier in the gut by investigating how mucus is produced, modified and released. We hypothesise that SOS can reduce obesity and associated health problems by regulating mucus and gut bacteria. Methods: Accordingly, HFD-fed mice were used in this study. Results: The results showed that SOS alleviated HFD-induced weight gain and glucose disorders. It also enhanced the gut mucus barrier by promoting goblet cell differentiation and regulating mucus-related genes. In addition, SOS intervention was associated with increased abundance of potentially beneficial gut taxa. These bacterial changes were linked to better health measures. In conclusion, our findings demonstrate that SOS confer metabolic protection against HFD-induced obesity, at least partially, by coordinately modulating the mucus–microbiota axis. Conclusions: These data suggest that SOS may alleviate obesity and related disorders by improving the intestinal mucus layer and gut microbiota. Full article

Cow’s milk allergy (CMA) remains one of the most common food allergies in infancy, requiring the avoidance of cow’s milk and its derivatives. Breast milk is the best source of nutrition for infants. For those infants with CMA whose mothers are unable to breastfeed or choose not to, extensively hydrolysed formulas (eHFs) are widely recommended as first-line milk substitutes, whereas hydrolysed rice formulas (HRFs) are increasingly recognised as a viable alternative. This concept paper provides a healthcare professional (HCP) perspective on HRF, drawing on expert consensus from two meetings convened in 2025. Discussions noted the long history of safe and effective HRF use in Europe, its nutritional adequacy, and the evolving international guidelines supporting HRF as an alternative first-line option. A key meeting outcome was the development of a practical decision tree to help UK clinicians decide when HRF should be the preferred choice. Key considerations for its use in non-breastfed infants include the following: parental/caregiver stress related to persistent symptoms; ongoing symptoms despite multiple interventions; cultural and lifestyle choices; religious dietary requirements; and specialists’ recommendations. Secondary considerations highlighted by HCPs include the following: proven reactions whilst infants are breast-milk-fed together with parental request for formula; faltering growth; multiple symptoms; taste acceptance (older infants); and parental preference based on experience. The role of functional components, such as prebiotics and human milk oligosaccharides (HMOs), was noted in regard to the emerging evidence of benefits to the microbiome and immune development. The experts emphasised the importance of engaging HCPs across all levels of CMA care and addressing challenges in translating current guidance into treatment practice. It was concluded that, overall, HRF represents a nutritionally complete, plant-based alternative that has been shown to be well tolerated (taste, symptoms) in clinical studies. It can be used to broaden therapeutic options for infants with CMA in the UK who are not exclusively fed breast milk. Full article

This study investigates the extraction of pectic polysaccharides from rapeseed meal (RSM) using both conventional and enzyme-assisted techniques, and the obtained pectic polysaccharide fractions will be used later to produce prebiotic pectic oligosaccharides (POS). A two-step process was developed, involving enzymatic treatment with Alcalase^® 2.4 L for 2 h and Cellic^® CTec3 HS preparations for 24 h, followed by ammonium oxalate extraction, which effectively isolated two pectic polysaccharide-enriched fractions: PP-EAE (first step) and the resulting Ca-bound pectic polysaccharides fraction (CaPP-EAE) (second step). Both fractions exhibited a bimodal molecular weight profile, indicative of the presence of long-chain polysaccharides alongside oligosaccharides. CaPP-EAE compositional analysis revealed that the fraction contained 56.8% galacturonic acid (GalA), low methyl-esterified (LM) pectins with 53.2% homogalacturonan (HG) and 30.2% rhamnogalacturonan I (RG-I) domains, featuring side chains of arabinan, arabinogalactan, and galactan. Subsequent enzymatic treatment with 0.5% (v/v) of Pectinex^® Ultra Passover for 30 min transformed these fragments into a mixture of short-chain POS. Importantly, the produced short-chain POS fraction demonstrated enhanced prebiotic activity, particularly for bacterial strains of the family Lactobacillaceae, compared to a yeast strain. These findings provide a sustainable, biorefinery-compatible approach for extracting and modifying RSM polysaccharides, supporting the development of structurally defined POS as novel prebiotics. Full article

Background: Gut microbiome-derived metabolites, particularly short-chain fatty acids (SCFA) and tryptophan derivatives, are central mediators of the gut–brain axis. This ex vivo study assessed how nutritional interventions impact such metabolites during early life, a critical period for neurodevelopment. Methods: The effects of galacto-oligosaccharides (GOS), nutrient blends (vitamins, minerals and amino acids) and their combinations were evaluated in the gut microbiomes of infants (2–4 months, n = 6) and young children (2–3 years old, n = 6) using the ex vivo SIFR^® technology. Results: Baseline microbiome composition was age-dependent, with infants displaying lower α-diversity and greater interpersonal variability. After ex vivo incubation, nutrient blends increased the propionate/butyrate ratio and branched-chain fatty acids in young children and elevated several B-vitamins and amino acid-derived metabolites, including indole-3-carboxaldehyde, imidazoleacetic acid and pipecolinic acid. Combining nutrient blends with GOS exhibited potential synergistic effects on propionate (infants) and 2-hydroxyisocaproic acid (HICA, both age groups). GOS strongly stimulated Bifidobacteriaceae and increased metabolites linked to bifidobacterial metabolism like acetate, HICA, N-acetylated amino acids, aromatic lactic acids and acetylagmatine; in young children, butyrate and γ-aminobutyric acid (GABA) also increased. Conclusions: Combinations of GOS with nutrient blends impacted microbiome-derived metabolites associated with the gut–brain axis, with potential synergistic increases of metabolites with emerging roles in neurodevelopment, including GABA, acetylagmatine and HICA. Despite shared bifidogenic effects, differences between age groups indicate that microbiome maturity may influence responses to nutritional intervention. Future clinical studies are needed to determine whether these metabolite changes translate into neurodevelopmental benefits in vivo. Full article

N-glycosylation is a fundamental post-translational modification that contributes to protein folding, stability, and secretion in eukaryotes. The catalytic subunit STT3 of the oligosaccharyltransferase complex mediates the transfer of preassembled oligosaccharides to nascent polypeptides in the endoplasmic reticulum. Here, we identified and functionally characterized PsSTT3B, one of the STT3 paralogs in Phytophthora sojae (P. sojae). PsSTT3B plays an important role in the growth, development, and pathogenicity of P. sojae. CRISPR/Cas9-mediated deletion of PsSTT3B resulted in reduced vegetative growth, sporangia production, and zoospore production in P. sojae. PsSTT3B deletion mutants demonstrated significantly reduced virulence on soybean leaves and etiolated seedlings. Importantly, PsSTT3B deletion mutants also exhibited reduced zoospore germination and diminished chemotaxis toward soybean isoflavones. Moreover, deletion of PsSTT3B increased sensitivity to tunicamycin and dithiothreitol and influenced the ConA-binding glycoprotein profile. Our findings show that PsSTT3B is associated with asexual development, virulence, and sensitivity to ER stress-related conditions of P. sojae. Our study suggests that PsSTT3B represents a potential candidate gene for the prevention and control of P. sojae. Full article

Cyclodextrins (CDs) have gained increasing attention as versatile platforms for enhancing drug delivery to the central nervous system, particularly in overcoming the restrictive properties of the blood–brain barrier (BBB). Owing to their unique cyclic oligosaccharide structure, CDs are capable of forming inclusion complexes with a wide range of therapeutic agents, thereby improving their solubility, stability, and bioavailability. In addition to their role as excipients, growing evidence indicates that CDs can actively modulate biological processes, including membrane fluidity and cholesterol homeostasis, which are critical factors in neurological disorders. This review explores the application of CDs in facilitating drug transport across the BBB through multiple mechanisms, including carrier-mediated transport, receptor-mediated transcytosis, and nanoparticle-based delivery systems. Special emphasis is placed on their use in the treatment of neurodegenerative and neurological diseases, such as Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, Niemann–Pick type C disease, and other central nervous system disorders. In these contexts, CD-based formulations have demonstrated the ability to enhance brain targeting, reduce pathological protein aggregation, and improve therapeutic outcomes in preclinical models. This review uniquely integrates cyclodextrin’s physicochemical properties with specific blood–brain barrier transport mechanisms, proposing a structure–transport–therapy framework that enables a more predictive understanding of brain-targeted drug delivery. Full article

Chickpea flour represents a valuable plant-based ingredient due to its high protein and fiber content; however, its application is limited by antinutritional factors and off-flavor compounds. Fermentation with LAB and yeasts, applied individually or in consortia, resulted in significant microbiological, nutritional, and aromatic changes. The fastest acidification (pH 3.9) and the most effective control of Enterobacteriaceae (<4 log CFU/g after 48 h) were observed in samples containing Lactiplantibacillus plantarum LP23, both as a monoculture and in combination with Debaryomyces hansenii Y15A. Peptide content significantly increased in all fermented samples compared to the control, with a synergistic effect in the co-culture Yarrowia lipolytica Y3 + Lacticaseibacillus paracasei L (around 230%). A pronounced reduction in raffinose-family oligosaccharides was observed, especially in the consortia Y. lipolytica Y3 + Lcb. paracasei L and D. hansenii Y15A + Lacp. plantarum LP23 (0.11–0.16 mmol/100 g). Samples with lower total volatile levels showed higher olfactory acceptability due to a marked reduction in aldehydes (up to 70–95% vs. control), and a balanced accumulation of alcohols, esters, ketones, and organic acids. Overall, LAB–yeast consortia effectively enhanced the nutritional quality, safety, and sensory properties of chickpea flour, supporting its use as a functional ingredient in plant-based foods. Full article

Hyaluronan (HA) is a major extracellular matrix glycosaminoglycan essential for tissue integrity, immune homeostasis, and host defense. Many microbial pathogens exploit host HA by producing hyaluronidases (Hyls), enzymes that degrade HA to promote tissue invasion, nutrient acquisition, immune modulation, and biofilm formation. Unlike mammalian Hyls, microbial Hyls predominantly function as β-elimination lyases, generating unsaturated disaccharides and oligosaccharides with distinct biological activities. Recent mechanistic and structural insights reveal that distinct microbial Hyl variants uniquely shape host–microbe interactions and disease outcomes. This review focuses on microbial Hyls, specifically bacterial Hyls, emphasizing their roles in host immune regulation and inflammatory diseases, particularly in Cutibacterium acnes-mediated acne pathogenesis. We also discuss emerging therapeutic strategies targeting the HA-Hyl axis to modulate inflammation, highlighting their potential as a foundation for novel human therapeutics. Full article