Food Chemistry

Why was the work done?: Gluten-free (GF) beers typically exhibit poorer foam stability than conventional barley malt-based beers, yet the molecular basis underlying this difference remains poorly understood. This study aimed to investigate protein-level factors associated with foam stability in GF and barley beers. How was the work done?: Three commercially produced GF beers, comprising two lagers brewed from rice or sorghum, and one pale ale brewed from a millet-buckwheat-rice blend, were compared with two commercially produced barley-malt beers, comprising a lager and a pale ale. Soluble protein was isolated from freeze-dried beer powder using TCA-acetone. Protein profiles were examined using SDS-PAGE, and proteomes were characterised and compared using label-free quantitative LC-MS/MS. What are the main findings?: GF beers showed lower soluble protein concentrations and significantly lower foam stability than the barley beers. SDS-PAGE revealed fewer and weaker protein bands in GF beers compared with barley beers. Proteomic analysis showed that barley beers were enriched in known foam-active proteins, particularly lipid transfer protein 1 (LTP1) and Protein Z (serpin family members), whereas these proteins were not detected or were detected only at very low levels in GF beers.

Cloudberry (Rubus chamaemorus L.)-derived nanovesicles (NVs) represent a promising but still poorly characterized class of plant-derived vesicles with potential relevance for skin-related applications. Here, we isolated cloudberry fruit-derived NVs and investigated their physicochemical and molecular properties, cellular uptake, cytocompatibility, and functional effects in human dermal fibroblasts (HDF) and HaCaT keratinocytes. Nanoparticle tracking analysis and transmission electron microscopy confirmed a nanosized vesicle preparation with characteristic round morphology, while protein quantification supported reproducible isolation of NV-associated material. In vitro, cloudberry NVs showed concentration-dependent effects on cell viability and proliferation, with lower doses being better tolerated. Labelled NVs were internalized by both HDF and HaCaT cells in a time-dependent manner. Under oxidative stress conditions, cloudberry NVs reduced H2O2-induced senescence-associated {beta}-galactosidase staining in HDFs and exerted cytoprotective effects in both cell lines, alongside measurable cell-free antioxidant activity in the DPPH assay. In scratch wound-healing assays, cloudberry NVs modulated wound closure in a dose-dependent manner, with the lowest tested concentration showing the most favorable response. UHPLC-MS/MS-based proteomics and metabolomics further indicated the presence of diverse secondary metabolites and stress-related protein cargo. Together, these results support the view that cloudberry-derived NVs are biologically active plant nanovesicles with potential utility in skin-related regenerative applications. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=126 SRC="FIGDIR/small/741293v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@c9094dorg.highwire.dtl.DTLVardef@81a1b5org.highwire.dtl.DTLVardef@9fafe2org.highwire.dtl.DTLVardef@1d421d3_HPS_FORMAT_FIGEXP M_FIG C_FIG

Kakadu plum (Terminalia ferdinandiana), an Australian native fruit, is among the richest known dietary sources of vitamin C and hydrolysable tannins, yet its capacity to protect the intestinal epithelium against oxidative stress remains largely unexplored. This study optimised the extraction of bioactive compounds from freeze-dried Kakadu plum powder and evaluated their antioxidant activity using both chemical and cellular antioxidant in vitro assay. Phenolic compounds were extracted using three solvents (water, 80% ethanol, and 80% methanol) combined with shaking, ultrasound, or microwave assistance. Solvent, rather than processing technique, was the dominant determinant of antioxidant capacity: ethanol and methanol maximised total phenolic content, total flavonoid content, and DPPH radical-scavenging activity, whereas water extracts showed the highest ferric-reducing antioxidant power. Twenty-four phenolic compounds identified by HPLC-ESI-QTOF-MS/MS were mapped by network pharmacology to nine core oxidative-stress targets, and cross-species molecular docking predicted conserved binding of key phenolics to canine orthologs of PTGS2 and MMP2. In an H2O2-induced oxidative-stress in vitro cell model using canine small intestinal epithelial cells, both water (less than 25 ug/mL) and ethanol (less than 250 ug/mL) extracts significantly suppressed intracellular reactive oxygen species (ROS) in a dose-dependent manner, with the ethanol extract effective across a wider concentration range. This work demonstrated that Kakadu plum extract could be a promising natural, multi-target antioxidant ingredient for canine intestinal health, and provided a reference for future in vivo research.

Knowledge of the influence of particle size, blending, and processing temperature on flour functionality is critical for the utilisation of emerging grains. This study investigated the flour pasting behaviour of four Australian native grains - Button Grass, Curly Mitchell Grass, Native Millet, and Weeping Grass - in relation to particle size, wheat flour substitution at various levels (0-30%), and elevated RVA temperatures (95-140 degrees Celcius). The pasting properties of flour fractions (fine, medium, and wholegrain) were analysed using a Rapid Visco Analyser, and water absorption capacity was also determined. Particle size effects were species-specific. Native Millet and wheat had higher viscosity in the finer fractions, whereas Weeping Grass exhibited maximum viscosity in the medium fraction. Button Grass and Curly Mitchell Grass showed limited sensitivity to particle size, indicating a stronger influence of intrinsic structure. Increasing substitution with native flours generally reduced viscosity, except for Weeping Grass, which was less affected and maintained stable viscosity. High-temperature rapid visco analysis revealed distinct thermal responses: most grains showed reduced final viscosity at elevated temperatures, whereas Weeping Grass maintained relatively high final viscosity across most temperatures. These findings demonstrate the diverse functional behaviour of Australian native grains and provide insights into their potential application in gluten-free and composite flour systems.

Nisin is a promising antimicrobial peptide widely used in food preservation due to its efficacy against Gram-positive spoilage and pathogenic bacteria. Although Nisin is increasingly applied in the food sector, the biopeptide suffers from instability within food matrixes and can rapidly lose its antimicrobial potential following interaction with food biomolecules. Thus, it is necessary to investigate approaches that can be employed to extend the stability and activity of Nisin. Hence, the aim of this study was to develop and characterise a chitosan-alginate polyelectrolyte microencapsulation system capable of enhancing Nisin stability while retaining antimicrobial activity. The microencapsulation of Nisin was achieved by pre-gelation of alginate using calcium chloride and subsequent direct electrostatic interaction between cationic Nisin and chitosan with pre-gelled anionic alginate at pH 5.0. Following microcapsule formation, physicochemical and structural characterisation was performed using Zeta potential determination and measurement of the polydispersity index (PDI) via dynamic light scattering. SEM micrographs were used to confirm morphology, while Fourier-transform infrared (FTIR) spectroscopy and high-performance liquid chromatography (HPLC) were utilised to assess chemical integrity and functional group preservation of encapsulated Nisin. Following this, stable microcapsules with diameters ranging from 150-200 nm and smooth surface morphology were obtained. Microcapsule formation was strongly influenced by formulation parameters, particularly pH, calcium ion concentration, and chitosan content, with deviations from optimal acidic conditions (< pH 5.0) resulting in aggregation, increased polydispersity, and reduced encapsulation efficiency. The microcapsules were monodispersed (PDI {approx} 0.30) and electrostatically stable, exhibiting a Zeta potential of approximately +36 mV. These microcapsules remained stable over a prolonged storage period of 21 days under refrigerated conditions while retaining antimicrobial activity against Bacillus cereus. Encapsulation efficiency reached approximately 65%, confirming effective retention of Nisin within the polymer matrix. Overall, the findings demonstrate that chitosan-alginate ionic gelation is a non-denaturing and effective encapsulation strategy for extending the functional stability of Nisin. These microcapsules show strong potential as natural antimicrobial delivery systems for food and beverage applications, particularly in acidic food matrices, with implications for improved food safety and shelf-life extension.

Traditional Mongolian fermented foods have been extensively utilized for dietary regulation and the promotion of gastrointestinal health. However, spontaneous fermentation remains inherently unpredictable, leading to significant variations in microbial community dynamics, metabolite accumulation, and the consistency and quality of the final product. Drawing on the traditional preparation of Mongolian acidic foods, this study established a controlled production strategy for whole-wheat probiotic fermented soup (WWPFS) by combining enzymatic pretreatment with probiotic-directed fermentation. Physicochemical characterization, 16S rRNA gene-based microbial community profiling, LC-MS/MS-based untargeted metabolomics, safety evaluation, and an Escherichia coli-induced gut microbiota dysbiosis model were employed to optimize and comprehensively characterize the fermentation process of WWPFS. The optimized process established a reproducible fermentation system consistently dominated by Lactobacillus and Bacillus across independent fermentation batches. Compared with traditional spontaneous fermentation, probiotic-directed fermentation remodeled the physicochemical properties of the whole-wheat matrix, including carbon, nitrogen, phosphorus, sulfur, and mineral composition, and facilitated the accumulation of putatively annotated LC-MS/MS features, including DL-lactate, 1,4-D-xylobiose, diacetyl, and phenyllactic-acid-related features derivatives. Acute oral and 28-day repeated-dose toxicity evaluations showed no treatment-related adverse effects within the tested dose range and study duration. In the Escherichia coli-induced gut microbiota dysbiosis mouse model, microbial richness, diversity, and community structure differed among the experimental groups, and both low- and high-dose WWPFS groups showed significant shifts in overall gut microbial community composition relative to the model group after multiple-testing correction, together with directional recovery of selected model-responsive bacterial genera. Cross-system integration identified coordinated response patterns between fermentation-derived metabolite features and model-responsive gut bacterial taxa, supporting a potential metabolite-microbiota link in WWPFS-mediated gut microbiota modulation. In summary, probiotic-directed fermentation improved the controllability of the traditional Mongolian fermented food production process, reshaped its metabolic profile, and enhanced its potential to modulate the gut microbiota. These findings provide experimental evidence supporting the modernization of traditional Mongolian fermented foods and the development of probiotic-based functional foods.

This study aims to investigate changes in protein secondary structures (-helix, -sheet, random coils, and -turn) and macronutrient distribution in different cool-season oat varieties and steam-pressure toasting durations using synchrotron-based mid-infrared (Mid-IR) spectroscopy and Fourier Transform Infrared spectroscopy (FTIR) imaging. All oat samples, provided by the Crop Development Center at the University of Saskatchewan, were harvested over three consecutive years (2018, 2019, and 2020). The first experiment compared four oat varieties (CDC Arborg, CDC Nasser, CDC Haymaker, and Summit), while the second examined CDC Nasser oats subjected to steam-pressure toasting (SPT) at 121 for 0, 30, 60, 90, and 120 minutes. FTIR chemical imaging revealed that carbohydrates, proteins and lipids in the four oat varieties were mainly concentrated in the endosperm, aleurone layer and embryo, crease region, and remained unchanged after SPT. Peak-fitting deconvolution of the Amide I band (1700-1600 cm-1) and subsequent quantitative analysis revealed that the four oat varieties exhibited broadly similar protein secondary structure profiles, with statistically significant but subtle variety effects detected for -helix (P = 0.026), -turn (P = 0.047), and the -helix to -sheet ratio (P = 0.048), however, -sheet and random coil proportions did not differ significantly among varieties. In contrast, SPT induced pronounced structural rearrangements, with significant increase in -sheet proportion (P = 0.003) and significant decreases in random coil content (P = 0.026). Notably, 30 minutes of toasting was sufficient to significantly increase -sheet and decrease the random coil contents. These changes are consistent with heat-induced protein denaturation and intermolecular -sheet aggregation, where thermal energy breaks the hydrogen bonds that stabilize the disordered random coil conformation, causing the unfolded polypeptide chains to reassemble into highly ordered -sheet aggregates. After SPT, the peak centers of Amide I and II bands shifted to lower wavenumbers and both bands broadened while their intensities were maintained, reflecting the reorganization of the remaining protein into -sheet aggregates rather than any loss of amide-active protein. These findings suggest that, although genotype has a relatively minor effect on the protein secondary structure of oats, hydrothermal treatments fundamentally reorganize the protein matrix from a disordered to an ordered conformation, which may have implications for protein digestibility, solubility, and nutritional function.

Background: Natural compounds from avocado fruit (avocadene, avocadyne, and acetate derivatives) exhibit notable biological activity, although their molecular mechanisms remain unclear. The avocado-derived lipids exert potent nematocidal activity against several parasitic nematodes. In Caenorhabditis elegans (C. elegans), those compounds caused concentration-dependent toxicity, impairing first stage larval growth, egg hatching, and adult survival. Treated worms exhibited impaired mitochondrial respiration, reduced oxygen consumption, and elevated reactive oxygen species. These effects suggest that avocado lipids disrupt mitochondrial function and lipid metabolism, in part by inhibiting acetyl-CoA carboxylase, the rate-limiting enzyme of fatty acid biosynthesis. Methods: We investigated the effects of these compounds on the lipid profile of C. elegans and their association with endogenous lipid pools using NMR spectroscopy, click-chemistry-based fluorescence labeling, thin-layer chromatography (TLC), and microscopy. Results: Lipidomic analysis of stage 4 larvae (L4) and embryos treated with avocadene acetate revealed increased lipid NMR signals. Fluorescence-assisted TLC and NMR further suggested that avocadyne preferentially associates with triglyceride-linked fatty acids, particularly monounsaturated and flexible polyunsaturated chains, without detectable interactions with conformationally-constrained polyunsaturated species. Fluorescent avocadyne derivatives were efficiently internalized with distinct localization patterns in L4 larvae and embryonic cells. Conclusions: Overall, the lipid homeostasis remodeling of L4 larvae in response to lipotoxic shock was associated with phospholipid increase and remarkable lipid droplets onset, whereas embryos showed accumulation of lipids in enlarged droplets and developmental arrest.

Type 2 diabetes (T2D) is a prevalent metabolic disorder affecting millions worldwide, characterized by insulin resistance and impaired glucose homeostasis. While mammalian models are widely used, Drosophila melanogaster provides a powerful alternative due to its conserved insulin signaling pathways, genetic tractability, and suitability for high throughput studies. In addition to glucose dysregulation, lipid metabolism plays a crucial role in T2D pathophysiology, as alterations in lipid composition contribute to insulin resistance and metabolic dysfunction. Lipidomic studies have emerged as an essential approach to identify metabolic signatures and potential biomarkers for disease progression and therapeutic targeting. In this study, T2D like model was established by inducing insulin resistance through knockdown of the insulin receptor in brain insulin-producing cells using the dilp2-Gal4>UAS-InRRNAi system. This genetic manipulation resulted in significant metabolic dysregulation, including elevated glucose, trehalose, and triacylglyceride levels, along with increased oxidative stress indicators. Additionally, mRNA expression analysis of key insulin signaling components, including insulin receptor substrate 1, dilp2, dilp3, dilp5, and phosphorylated Akt, further validated the model. To further investigate metabolic alterations, Lipid profiling was performed using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) in non targeted LC-MS-based metabolomics approach to identify lipid biomarkers associated with T2D. Multivariate statistical analyses, including PCA and PLS-DA, revealed distinct lipid signatures between wild-type and T2D flies. Notably, specific phosphatidylglycerol species PG 34:0, PG 34:4, PA 38:3, PIP 38:1, PIP2 38:6, and LPS 24:0 demonstrated an area under the curve (AUC) of 1, indicating their strong reliability as lipid biomarkers for T2D diagnosis.

Ensuring the safety and suitability of proteins considered for food and biotechnology applications is an important challenge. Computational approaches can facilitate the prioritisation of candidate proteins for further investigation, particularly given the rapid growth of genomic and metagenomic sequence datasets. In this study, we developed computational models to distinguish edible plant tissue-associated (EPTA) proteins from non-EPTA proteins. The positive class comprised 939 proteins with experimentally validated evidence of expression in plant tissues commonly consumed by humans, hereafter referred to as EPTA proteins. The negative class comprised 939 proteins annotated in UniProt as toxic, allergenic, or antinutritional, hereafter referred to as non-EPTA proteins. Models were evaluated using five-fold cross-validation with a cluster-based partitioning strategy, ensuring that no protein in the validation set shared more than 40% sequence identity with proteins in the corresponding training set. First, composition-based sequence features were evaluated to establish a baseline for EPTA or non-EPTA classification, with the Extra Trees (ET) classifier achieving a validation AUROC of 0.848. To incorporate evolutionary information beyond amino acid composition, PSSM profiles were subsequently evaluated, yielding an AUROC of 0.842 with an SVM classifier. Finally, to capture richer sequence patterns and contextual information using protein language modelling, we evaluated ESM2-based models. The fine-tuned ESM2-T33 model achieved the highest validation AUROC of 0.925 and was selected as the final predictive model. The resulting approach was implemented as the EdiProPred web server (https://webs.iiitd.edu.in/raghava/edipropred/) and standalone software for computational classification of plant proteins. The EdiProPred framework provides a useful computational tool for prioritising candidate plant proteins and facilitating the exploration of novel protein resources for food and biotechnology applications.

Background and AimsThe genus Solanum including the wild species S. dulcamara, is rich in specialized metabolites such as steroidal glycoalkaloids (SGAs). Yet, much of this chemical diversity remains poorly characterized. This study aims to provide a comprehensive assessment of intra-specific chemodiversity in S. dulcamara. Using a dataset generated from 42 globally distributed accessions, we tested whether metabolic profiles differ among plant organs. We postulated that metabolic richness and abundance vary across accessions. Additionally, we hypothesized that differences in geographic origin or altitude affect SGA chemodiversity. MethodsAn untargeted metabolomic approach was applied to leaf, flower and root samples of 42 S. dulcamara accessions. Plants were grown in the greenhouse, and the extracted metabolites were analyzed using UHPLC-HRMS/MS in positive and negative ionization modes. Data processing and metabolite annotation were performed with a tailored bioinformatics workflow. Multivariate analyses were performed to evaluate chemical variation across organs and accessions. Key ResultsOur analyses revealed both organ and accession-specific metabolic diversity. Principal component analysis and clustering analyses revealed metabolic differentiation between leaves, flowers and roots. Leaves showed the highest metabolite richness and abundance, while roots showed the lowest. Alkaloids, especially SGAs, dominated positive mode profiles in roots, whereas shikimates and phenylpropanoids were prominent in negative mode profiles. Based on the leaf and flower SGAs profiles, four chemotypes were identified. Analyses of flavonoid and cinnamic acid derivatives, however, did not reveal chemotypes. Feature-based molecular network analyses confirmed that metabolite clusters are associated with plant organs, but not with altitude or geographic origin of the accessions. ConclusionsThe intraspecific chemodiversity within S. dulcamara is mainly driven by organ and accession-specific metabolic differences. We identified four SGA leaf and flower chemotypes, suggesting possible functional and ecological roles of this aboveground chemodiversity. These insights may contribute to applied research in plant resistance breeding and crop production.

This study investigates electro-fermentation candida Tropicalis SY005 to address fermentation kinetics limitation during grape must fermentation. In comparison with non-stimulated control sample, EF substantially enhanced sugar depletion, TSS drop by day 3 and generated a strongly reduced state (ORP -100 to -143mV). The oxidation-reduction shift enhanced cellular NAD+ regeneration, reducing total fermentation duration from 264 h to 72 h. GC-MS analysis showed pronounced major characteristic volatile compound confirming substantial metabolic pathway shifts in flavor of glycolytic flux. Moreover moderate electric field promoted cellular membrane electropermeabilization substantially promoting bioactive extraction.

Branched-chain amino acids (BCAAs) are essential amino acids involved in protein synthesis and energy metabolism. Because animals cannot synthesize BCAA de novo, plant-derived BCAAs are important to human nutrition. Although mungbean sprouts are widely consumed as functional plant-based food materials, changes in individual BCAA accumulation and their transcriptional regulation during mungbean germination remain poorly understood. In this study, amino acid contents and transcriptomic profiles were analyzed at three germination stages, 8H, 24H, and 72H. Total BCAA content increased during germination, whereas individual BCAAs exhibited distinct temporal accumulation patterns. Isoleucine and valine increased until 72H, while leucine increased during early germination and decreased after 24H. Transcriptome analysis revealed time-dependent expression changes in BCAA biosynthesis and degradation genes associated with the leucine decrease after 24H. These findings suggest that 24H represents an important transition point for BCAA accumulation and compositional change during mungbean germination. This study provides molecular evidence for the regulation of BCAA metabolism during mungbean germination and supports the potential use of germinated mungbean as a plant-based amino acid resource.

Apple scar skin disease (ASSD), caused by Apple scar skin viroid (ASSVd), is characterized by peel scarring, cracking, dappling, and fruit deformation, resulting in reduced fruit quality and marketability. Despite its economic importance, the physicochemical and metabolic alterations underlying disease progression remain poorly understood. To address this knowledge gap, apple fruits representing four stages of ASSD (healthy, lightly infected, moderately infected, and highly infected) were comprehensively characterized. ASSVd infection was confirmed by RT-PCR, amplicon sequencing, and phylogenetic analysis. Fruit morphology and quality attributes, including firmness, total soluble solids (TSS), pH, titratable acidity (TA), and total phenolic content (TPC), were evaluated. ASSVd infection significantly reduced fruit weight and firmness and altered TSS and TA, indicating progressive deterioration of fruit quality. To investigate the underlying metabolic changes, peel and pulp tissues were analysed separately using gas chromatography-mass spectrometry (GC-MS), while major soluble sugars were quantified by 1H nuclear magnetic resonance (1H NMR) spectroscopy. Integrated metabolomic analyses revealed distinct tissue-specific metabolic reprogramming during disease progression. Major soluble sugars declined significantly during early infection, followed by tissue-dependent recovery at later stages, whereas organic acids, amino acids, phenolics, lipids, polyols, and pentacyclic triterpenoids exhibited dynamic stage-dependent changes. Notably, lupeol accumulated progressively, whereas ursolic acid and oleanolic acid declined, indicating disease-associated alterations in host triterpenoid metabolism. Multivariate analyses demonstrated clear metabolic separation among disease stages. Lupeol, ursolic acid, and chlorogenic acid were identified as candidate discriminatory metabolites in the peel, whereas myo-inositol, chlorogenic acid, and aspartic acid were identified in the pulp. Collectively, these findings demonstrate that ASSD induces coordinated, tissue-specific physicochemical and metabolic reprogramming that compromises postharvest fruit quality and reshapes defence-associated metabolism. This study provides the first integrated metabolomic characterization of ASSD progression and identifies potential metabolic biomarkers for disease diagnosis and severity assessment. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=142 SRC="FIGDIR/small/741181v1_ufig1.gif" ALT="Figure 1"> View larger version (56K): org.highwire.dtl.DTLVardef@92e453org.highwire.dtl.DTLVardef@1cc2436org.highwire.dtl.DTLVardef@15d7e27org.highwire.dtl.DTLVardef@1056568_HPS_FORMAT_FIGEXP M_FIG C_FIG

High dietary fructose consumption is a major contributor to the development of obesity and related cardiometabolic diseases, highlighting the need for accurate assessment of fructose metabolism in humans. Stable isotope tracer approaches, such as 13C6-fructose, require highly sensitive and specific analytical methods to quantify both concentrations and isotopic enrichments. In this study, we developed and validated a robust gas chromatography-triple quadrupole mass spectrometry (GC-QQQ)-based method for the simultaneous measurement of unlabeled and 13C6-fructose in human plasma. The method employs oximation and per-acetate derivatization, and demonstrates high specificity and accuracy. Intra- and inter-assay precision were below 10%, with no detectable carry-over, and a lower limit of quantification (LLOQ) of 0.1 nmol/mL for concentration and 0.02 molar percent excess (MPE%) for enrichment and no interference from glucose. We further compared data acquisition using multiple reaction monitoring (MRM) and selected ion monitoring (SIM). MRM showed superior performance at the low concentrations and enrichment levels characteristic of clinical plasma samples, resulting in improved sensitivity and lower LLOQs compared to SIM. Overall, this validated method provides a sensitive and reliable approach for fructose tracer studies in humans. Its application will facilitate robust investigations into fructose metabolism, and its role in metabolic dysregulation and obesity-related disease.

Background: There is a growing availability of plant-based alternatives for bovine milk. However, plant-based drinks differ e.g. in protein quantity, protein quality and carbohydrate composition. Knowledge on the impact this has on digestion-related outcomes such as postprandial amino acid (AA) availability and gastric behavior is still limited. Additionally, data on milk alternatives with higher overall protein concentration are lacking. Objective: We aimed to compare the differences in postprandial AA profiles, glycemic response, and gastric behavior between bovine milk and plant-based drinks with or without added protein using representative commercial drinks. Methods: 12 healthy males participated in a randomized crossover study with three treatments. After an overnight fast, participants drank 750 ml of semi-skimmed milk (Milk), a standard oat-based drink (OBD) or an oat-based drink enriched with pea protein (OBD+Pea). Blood samples were taken for measurement of plasma AA, glucose, insulin and FGF21 over 5 hours. Gastric content changes were assessed using magnetic resonance imaging scans for Milk and OBD+Pea over 2 hours. Results: Compared with Milk, both oat-based drinks elicited lower 5-h total AA iAUC (OBD -169706; OBD+pea -139552 microM*min; both P<0.001) and EAA iAUC (OBD -91748; OBD+pea -78199; both P<0.001), with no differences between oat-based drinks. Gastric content volume was lower for OBD+Pea versus Milk from 60-120 min (range -48 - -84 ml, all P<0.001). Compared to Milk, OBD and OBD+Pea increased glucose more from 15-45 min (range 0.86 - 1.73 mmol/l, all P<0.001) and insulin from 30-60 min (1.84 - 2.14-fold, all P<0.001). Conclusions: Protein enrichment in plant-based milk alternatives can be a valuable strategy to increase protein content and improve amino acid composition. However, such products still show faster gastric emptying, lower postprandial essential amino acid and higher glycemic and insulinemic responses compared to bovine milk because of their different carbohydrate and protein content and quality.

Biopreservatives including nisin and its derivatives are becoming more desirable in the food processing industry because of the growing demand for naturally preserved and minimally processed foods free from artificial preservatives. However, ensuring microbiological safety while meeting these consumer preferences remains a major challenge. This has necessitated the continuous investigation of potential new antimicrobial agents produced by naturally occurring microorganisms. Hence, this study explored the synthesis, characterisation, and optimisation of a bacteriocinogenic lactic acid bacterium and its antimicrobial product, possibly novel bacteriocin (Nisin 2A) from Lactococcus lactis isolated from commercial brined cheese. The isolation was achieved by screening for wild-type bacteriocin-producing lactic acid bacteria from dairy products using MRS media. Screening was performed using antagonism assays, yielding five producer organisms. Of these, the isolate whose metabolites exhibited the most potent antimicrobial activity was identified as Lactococcus lactis, which synthesised an active antimicrobial peptide designated as Nisin 2A, with a molecular mass of approximately 3.3 kDa as determined by UHPLC-MS and SDS-PAGE. Production of Nisin 2A was scaled up through fed-batch fermentation of Lactococcus lactis in modified MRS broth following process optimisation using a Plackett-Burman experimental design and purified by ammonium sulphate precipitation and solid-phase extraction (SPE). Furthermore, the antimicrobial potential of the bacteriocin was evaluated by the agar well diffusion assay and quantified using the tube dilution method. The purified peptide demonstrated broad-spectrum antimicrobial activity, particularly against the test Gram-positive bacteria Bacillus cereus and retained its bioactivity across a wide pH range (3-9) and high thermal conditions (up to 100 {degrees}C). Furthermore, it had high sensitivity to proteolytic enzymes (Proteinase K and Trypsin). Notably, the peptide was thermostable and retained up to 90% of its initial activity after thermal treatment and maintained consistent inhibitory performance after extended storage. These findings highlight the potential application of Nisin 2A as a natural biopreservative in food systems.

Romanian bors, a traditional fermented wheat bran beverage, is produced through spontaneous fermentation and represents a complex microbial ecosystem. Despite its cultural importance and presumed health benefits, its microbial ecology and functional potential remain poorly characterized. The present study aimed to elucidate the microbial community structure of bors and link it to functional traits relevant to fermentation performance and food functionality by integrating culture-independent sequencing with culture-dependent isolation and functional characterization. A total of 32 bors samples (12 commercial and 20 homemade) were analyzed. Amplicon-based sequencing revealed a microbiome dominated by lactic acid bacteria (LAB), with lactobacilli accounting for the majority of the bacterial communities and Lactobacillus amylolyticus being identified as the most prevalent and abundant species. The yeast communities were mainly composed of fermentative taxa, including Pichia kudriavzevii and Kluyveromyces marxianus. Lactobacillus amylolyticus and P. kudriavzevii were also the most frequently isolated species among bacteria and yeasts, respectively. These results highlighted a strong adaptation of the microbial isolates to starch-rich cereal substrates and underscored the central role of these microorganisms in wheat bran fermentation for bors production. Whereas the sequencing-based analyses showed no significant differences in overall diversity between the commercial and homemade bors samples, the cultivation-based results indicated a higher bacterial richness in the commercial products. Notably, the culture-dependent method captured substantially fewer taxa, highlighting the complementary nature of the two approaches. Of a total of 101 bacterial strains (88 LAB and 13 acetic acid bacteria) isolated, many exhibited rapid growth and strong acidification capacity, reaching pH values below 4.5 within 12 h. A functional screening revealed that 21 % of these strains displayed -amylase activity, 65 % phytase activity, and 50 % {beta}-glucosidase activity, highlighting their capacity to metabolize cereal substrates and enhance the nutrient availability of bors. All strains showed antibacterial activity against at least one indicator bacterium tested, with a universal inhibition of Listeria monocytogenes. Overall, Romanian bors harbored a lactic acid bacteria-dominated core microbiome with a significant functional diversity. These findings underscored its potential as a rich source of functional and technologically important strains for application in starter and protective culture development.

The binding affinities of a chimeric analog of a riboflavin derivative linked to biotin, (6- (7,8-dimethyl-2,4-dioxo-3,4-dihydrobenzo[g]pteridin-10(2H)-yl)hexyl 5-((3aS,4S,6aR)-2- oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanoate), referred to as C6-Rf-biotin-tag, to the riboflavin binding retain or streptavidin are in the M range, 1.29 {+/-} 0.277 and 3.00 {+/-} 0.459, respectively. These values suggest that C6-Rf-biotin-tag has potential applications in diagnostic assay and labelling target flavin binding proteins. The C6-Rf-biotin-tag which was characterized with respect to physical and biochemical properties retains UV/Vis spectroscopic and fluorescence behavior similar to riboflavin.

Water-soluble vitamins are essential micronutrients that are not stored extensively in the human body and must therefore be replenished through dietary intake. Mungbean sprouts are widely consumed in some populations; however, their contributions as dietary sources of water-soluble vitamins and the extent of genotype-dependent variations remain insufficiently characterized. In this study, six water-soluble vitamins (B1, B2, B3, B6, B9, and C) were quantitatively and qualitatively analyzed using ultra performance liquid chromatography in sprouts derived from 34 mungbean genotypes. With the exception of pyridoxine (vitamin B6), all analyzed vitamins were detected across all genotypes, and their contents showed significant genotype-dependent variation. To determine whether differences in vitamin composition are reflected in functional responses, antioxidant capacity and enzyme-based bioactivity were evaluated. Antioxidant activity varied significantly among genotypes and showed a positive correlation with nicotinamide (B3) content, while tyrosinase inhibition was positively correlated with thiamine (B1). Evaluation of vitamin contents on a fresh-weight basis demonstrated that a single 104 g serving of mungbean sprouts can provide 46%- 206% of the recommended daily allowance of thiamine depending on genotype, while supplying all analyzed water-soluble vitamins, including B1, B2, B3, B6, B9, and C. This studys findings support the potential of mungbean sprouts to serve as plant-based dietary sources of water-soluble vitamins and offer a basis for selecting genetic resources to improve the vitamin-related nutritional quality of mungbean sprouts.