Search Results (157)

To enhance the gelling functionality of plant proteins, this study developed hybrid gels by blending casein with soy protein isolate (SPI) at various ratios using microbial transglutaminase (MTG) as a cross-linking catalyst. The gels were systematically characterized in terms of microstructure, water distribution, rheological and textural properties, secondary structure, and intermolecular interactions. Incorporation of casein significantly improved gel strength, water-holding capacity, and network uniformity. Notably, the 1:1 casein-to-SPI ratio yielded the highest performance, featuring extensive inter-protein cross-linking, an increased proportion of ordered secondary structures, and a finely porous matrix that effectively immobilized water. Mechanistically, MTG-catalyzed covalent bonding established the primary network scaffold, while hydrophobic interactions and disulfide bonds further stabilized the gel matrix. These findings demonstrate that MTG-induced Casein–SPI hybrid gels can enhance the functional properties of plant proteins and offer a viable strategy for designing sustainable protein-based food structures with tailored performance. Full article

This investigation focused on developing nutrient-dense Medjool date-based bars (MDBs) formulated with Medjool date paste, milk protein concentrate, whey proteins, and other functional ingredients. Comprehensive proximate analysis, mineral profiling, amino acid determination, and instrumental assessments, including color measurement, scanning electron microscopy, differential scanning calorimetry, and texture analysis, were performed, followed by organoleptic evaluation. Medjool date paste served as the primary carbohydrate source (76.44%), while whey protein isolate and milk protein concentrate contributed substantially to the protein fraction (89.26% and 81.62%, respectively). The resulting bars contained 19.32–26.78% crude protein, 10.96% fat, and 12.35–12.71% moisture, delivering 414.72–416.04 Kcal 100 g⁻¹. Sugar profiles remained consistent across formulations (total sugars: 36.77–36.98%), with appreciable mineral content including potassium (884–923 mg 100 g⁻¹), calcium (418–585 mg 100 g⁻¹), and phosphorus (402–459 mg 100 g⁻¹). The essential amino acid composition equaled or surpassed that of hen’s egg, establishing the product as a superior protein source. Antioxidant analysis demonstrated total phenolic content of 452.22–554.12 mg GAE 100 g⁻¹ and total flavonoids of 358.06–374.24 mg QE 100 g⁻¹, with consistent radical scavenging capacity, reduced browning via protein–polyphenol binding (ΔG −58 to −72 kJ mol⁻¹), a balanced texture (hardness 157–189 N), and consistent sensory scores (87.63–93.28% acceptability), without significant differences among formulations. Molecular docking confirmed β-lactoglobulin’s tight antioxidant shielding and caseinate’s flexible bioavailability boost, yielding shelf-stable functional snacks that advance date palm valorization. The results demonstrate the successful development of functional MDBs with an excellent nutritional profile and strong panelist acceptance. Full article

The present study evaluated the effects of sugarcane-derived polyphenol nutritional supplement on milk production, milk composition, and somatic cell count (SCC) in lactating Holstein cows. A second objective was to evaluate the quality and stability of yogurt produced with milk from supplemented cows. Sixteen cows (20 ± 2 L/d milk, 120 ± 21 days in milk, 550 ± 82 kg body weight) were assigned to either a control group or a polyphenol-supplemented group in a randomized block design for a six-week evaluation period. Milk yield and individual milk samples for composition analysis (fat, protein, lactose, non-fat solids, casein, free fatty acids, and milk urea nitrogen) were recorded weekly, and only SCC was monitored three times per week. Yogurt was produced from pooled milk of each treatment and evaluated for fermentation parameters (pH, titratable acidity), shelf life, and sensory attributes were evaluated using a triangle test (discriminative sensory analysis). Polyphenol supplementation did not significantly affect milk yield or composition (p > 0.05), but reduced SCC considerably (238 × 10³ vs. 593 × 10³ cells/mL; p < 0.01). Yogurt fermentation and storage parameters were not significantly affected by supplementation, although pH and titratable acidity changed considerably over time (p < 0.01). Sensory analysis indicated that only 37.3% of panelists correctly identified differences between yogurts, which did not differ from chance expectation in the triangle test. These results demonstrate that polyphenol supplementation can improve udder health, evidenced by reduced SCC, without compromising milk yield, composition, or yogurt quality. Polyphenols thus represent a promising nutritional supplement for enhancing dairy cow health and maintaining product quality. Full article

This study investigated the combined effects of oil concentration, emulsifier type, and saliva on the lubrication behavior of casein-based oil-in-water emulsions to support the design of milk-based foods with optimized mouthfeel. Emulsions stabilized with Tween 20, whey protein isolate (WPI), or sucrose ester were prepared at oil concentrations ranging from 0.01% to 3%, and their viscosity, microstructure, tribological properties, and ζ-potential were systematically characterized, with human saliva incorporated to simulate oral conditions. Oil concentration did not significantly alter viscosity, although droplet aggregation increased with higher oil levels. Lubrication performance was governed primarily by emulsifier type: Tween 20 generated an oil film at approximately 0.2% oil, WPI exhibited progressively enhanced lubricity with increasing oil concentration, and sucrose ester produced consistently poor lubrication due to its rigid interfacial layers. Saliva addition improved lubrication across all systems and reduced oil precipitation by promoting the formation of smaller, more stable structures. These findings demonstrate that emulsifier selection is central to modulating oil–protein–saliva interactions, with WPI at moderate oil levels yielding favorable lubrication with controlled oil release, thereby providing a mechanistic basis for developing healthier, palatable milk-based foods. Full article

The objectives of this study were (i) to analyze the ClassyFarm welfare scores in loose housing system (LHS) and tied housing system (THS) Rendena herds, and (ii) to investigate the influence of the housing on individual milk yield and quality traits in Rendena cows. The dataset consisted of 3761 individual milk samples from 750 Rendena cows, collected between August 2022 and November 2023 from 17 single-breed herds of the Veneto region. Available data included days in milk (DIM), parity, and milk yield, as well as fat, protein, casein, and lactose contents, somatic cell count, differential somatic cell count, and urea concentration. For milk traits, a linear mixed model included housing system, DIM, and parity as fixed effects, while cow, herd-test-date, and residual variability were random effects. In both housing systems, a good level of welfare was observed, although shortcomings in biosecurity measures were identified in both LHS and THS. Milk quality (in terms of protein % and casein %) and yield were higher in LHS compared to THS. The observed differences cannot be explained by the housing system alone, as other management and nutritional factors may have played a role, highlighting the need for further studies to clarify these contributions. Full article

Human embryonic kidney (HEK293) cells are a widespread choice for recombinant protein expression. To optimise yields, the hydrolysate Tryptone N1 (TN1) is commonly added post-transfection. TN1 is obtained by controlled enzymatic digestion of casein. As an animal by-product, TN1 faces stricter regulations during cross-country shipments than plant-based products. This raises the question of whether plant-derived peptides are a suitable alternative to TN1. Using polyethyleneimine (PEI) as a cationic polymer, we transfected HEK293-6E cells grown in suspension in serum-free medium and divided the transfectants into four groups (each in triplicate). Two plant-based hydrolysates each derived from pea and broad bean were compared with TN1 and a no-hydrolysate control group. We monitored the cultures for total cell numbers and viability at days 1, 4, and 5 post-transfection. Both plant-based hydrolysates and TN1 showed similar live cell percentages, in contrast to the no-hydrolysate control, which showed lower viability. Five days post-transfection, the expressed His-tagged protein, a tegumental antigen from the eukaryotic parasite Echinococcus granulosus, was retrieved from the serum-free culture supernatant, and the expressed recombinant protein was quantified. The linear ranges for the protein load on the stain-free blot and for the use of the fluorescent anti-His-Tag Alexa₄₈₈ antibody were determined. Using these parameters, stain-free Western blotting and total protein normalization were performed. The plant-derived pea and broad bean hydrolysates reproducibly resulted in similar expression levels as animal-derived TN1; all three hydrolysates were better than no hydrolysate. We conclude that plant-derived hydrolysates are a suitable, more sustainable replacement for TN1. Full article

Edible mushrooms are an underexplored source of industrial proteases, whose synthesis is highly dependent on the cultivation substrate. This study investigated the effect of nine culture media on the proteolytic profiles of Auricularia sp., Lentinus sp., Macrocybe sp., and Grifola frondosa. Fungi were cultivated on diverse media (e.g., Czapek, Malt, Soy Flour). We analyzed total protein, specific activities (total, cysteine, serine proteases) using a biochemical assay, and protein secondary structure via FTIR, with metabolic patterns identified by PCA. A dissociation was found between total protein yield (highest in MFI/Casein media) and specific activity (highest in maltose media), suggesting catabolite repression. Distinct metabolic strategies emerged: Grifola frondosa specialized in serine protease production in the minimal Czapek medium (catabolic derepression), while Macrocybe sp. maximized cysteine protease production on soy flour (substrate induction). FTIR confirmed this, revealing a β-sheet-dominant (75.5%) structure for Grifola extract versus a random-coil-dominant (60.8%) structure for Macrocybe. This study provides a framework for mechanism-based bioprocess design, enabling the tailored production of serine proteases from G. frondosa (Czapek medium) or cysteine proteases from Macrocybe sp. (soy medium) for customized biotechnological applications. Full article

The polymorphism of the bovine β-casein gene (CSN2) is of increasing interest due to its relevance for A2 milk production. This study genotyped 2773 Holstein-Friesian cows for five CSN2 alleles (A1, A2, A3, B, I) using both conventional DNA-based PCR and a newly evaluated direct PCR protocol. Eleven genotypes were detected, with A2/A2 (33.9%) and A1/A2 (30.3%) being the most common, resulting in an A2 allele frequency of 59.0%. Genetic diversity indices indicated moderate polymorphism and a significant deviation from Hardy–Weinberg equilibrium, consistent with ongoing selection for the A2 allele. Associations between CSN2 genotype and milk traits (305-day milk, fat, and protein yield; fat% and protein%) were evaluated using linear mixed-effects models including lactation number, age at calving, and calving year as covariates, and cow ID as a random intercept. Several genotype effects reached statistical significance (p < 0.05); however, all effect sizes were very small (partial η² < 0.01), indicating that any influence of CSN2 on production traits is negligible within this population and management context. These findings suggest that A2-oriented selection is unlikely to compromise productivity. The direct PCR genotyping method achieved 96–100% success and enabled substantially faster and more cost-efficient processing (approximately 80–90% reduction in reagent costs), providing a rapid and scalable approach for large herds. Full article

To promote sustainable food production, the effective valorization of agricultural byproducts is essential. This study investigated the potential of underutilized chestnut inner skin (CIS) and thinned young persimmon fruit (YPF) extracts as functional ingredients in pudding gels, selected as a complex model system utilizing heat-induced egg gelation with milk and sugar. Puddings were prepared by replacing water with 10% or 50% CIS or YPF extracts. We comprehensively evaluated the physicochemical properties (texture, color, viscosity), microstructure (SEM), and sensory quality. Additionally, immunoreactive allergenic proteins (ovalbumin, casein, β-lactoglobulin) were quantified using ELISA, and antioxidant activity was measured via DPPH and H-ORAC assays. Results indicated that while high extract concentrations (50%) negatively impacted texture by increasing hardness and forming air pockets, the 10% YPF treatment yielded a smooth, homogeneous microstructure comparable to the control. Crucially, the 10% YPF extract significantly reduced the concentration of detectable allergenic proteins, attributed to the formation of insoluble tannin–protein complexes, without compromising sensory acceptance. Furthermore, the addition of these extracts significantly enhanced the antioxidant activity of the puddings in a concentration-dependent manner. These findings demonstrate that 10% YPF is a promising candidate for developing sustainable, hypoallergenic, and antioxidant-rich functional food products. Full article

The aim of this study was to assess the influence of milk’s rennet coagulation properties (RCPs) on cheese yield and cheese-making losses in the production of Parmigiano Reggiano PDO cheese. Higher contents of citric acid (181.10 vs. 172.13 vs. 166.47 mg/100 g) and phosphorus (95.02 vs. 91.14 vs. 88.78 mg/100 g) in milk with optimal and sub-optimal RCPs, compared to milk with poor RCPs, respectively, positively affect the acidity of the milk, lowering the pH values (6.68 vs. 6.70 vs. 6.72, respectively), which results in a faster reaction between chymosin and casein and consequently a reduced time of milk coagulation. The lower values of curd firming time and the higher values of curd firmness, strength to cut (68.97 vs. 64.43 vs. 44.38 g), and strength to compression (31.48 vs. 30.49 vs. 25.70 g) for milk with optimal and sub-optimal coagulation, compared to milk with poor coagulation, result in a higher stress resistance across the technological steps of the cheese-making process, leading to lower fat losses (14.23 vs. 15.48 vs. 16.72%) in the whey and a higher cheese yield (8.79 vs. 8.56 vs. 8.08 kg/100 kg). Full article

Introduction: Magnetic nanoparticles are widely investigated as multifunctional platforms for drug delivery and theranostic applications, yet their biomedical implementation is hindered by aggregation, limited colloidal stability, and insufficient biocompatibility. Hybrid biopolymer coatings can mitigate these issues while supporting drug incorporation. Aim: This study aimed to develop casein–pullulan-coated Fe₃O₄ nanocomposites loaded with xanthohumol, enhancing stability and enabling controlled release for potential theranostic use. Methods: Fe₃O₄ nanoparticles were synthesized through co-precipitation and incorporated into a casein–pullulan matrix formed via polymer complexation and glutaraldehyde crosslinking. A 3² full factorial design evaluated the influence of casein:pullulan ratio and crosslinker concentration on physicochemical performance. Nanocomposites were characterized for size, zeta potential, morphology, composition, and stability, while drug loading, encapsulation efficiency, and release profiles were determined spectrophotometrically. Molecular docking was performed to examine casein–pullulan interactions. Results: Uncoated Fe₃O₄ nanoparticles aggregated extensively, displaying mean sizes of ~292 nm, zeta potential of +80.95 mV and high polydispersity (PDI above 0.2). Incorporation into the biopolymer matrix improved colloidal stability, yielding particles of ~185 nm with zeta potentials near –35 mV. TEM and SEM confirmed spherical morphology and uniform magnetic core incorporation. The optimal formulation, consisting of a 1:1 casein:pullulan ratio with 1% glutaraldehyde, achieved 5.7% drug loading, 68% encapsulation efficiency, and sustained release of xanthohumol up to 84% over 120 h, fitting Fickian diffusion (Korsmeyer–Peppas R² = 0.9877, n = 0.43). Conclusions: Casein–pullulan hybrid coatings significantly enhance Fe₃O₄ nanoparticle stability and enable controlled release of xanthohumol, presenting a promising platform for future targeted drug delivery and theranostic applications. Full article

Routine milk-recording data may provide valuable insights into dairy cow welfare, although their ability to accurately reflect herd-level welfare outcomes remains unclear. This study explored the associations between routinely collected milk biomarkers and farm-level welfare status using a comparative machine learning approach. Using the Welfare Quality^® (WQ^®) protocol, 43 commercial dairy farms were classified as Enhanced, Acceptable, or Not Classified. Farm-level milk variables included somatic cell count (SCC), differential somatic cell count (DSCC), fat-to-protein ratio (FPR), fat, protein, casein, lactose, urea, β-hydroxybutyrate (BHB), acetone, total plate count (TPC), and morning milk yield. Kruskal–Wallis tests revealed significant differences among welfare classes for DSCC, SCC, lactose, and milk yield (False Discovery Rate-adjusted p < 0.05). Six machine learning algorithms were trained using 10-fold stratified cross-validation. The Elastic-Net (ENET) model showed the highest mean performance (Accuracy = 0.72 ± 0.19; Kappa = 0.56 ± 0.31), followed by Random Forest and Multilayer Perceptron (Accuracy = 0.70). Model accuracy exhibited substantial variability across cross-validation folds, reflecting the limited sample size and class imbalance. Across models, the most influential variables were SCC, DSCC, lactose, milk yield, FPR, fat, and urea. Overall, the findings provide preliminary and exploratory evidence that routine milk biomarkers capture welfare-relevant patterns at the herd level, supporting their potential role as complementary indicators within data-driven welfare assessment frameworks. Full article

This study investigates the formulation and optimization of acid-stable water-in-oil-in-water (W/O/W) double emulsions stabilized by sodium caseinate (NaCN)–xanthan gum (XG) complexes, with the aim of developing a natural biopolymer-based delivery system exhibiting controlled release behavior. The emulsions were prepared at pH 4, and the effects of NaCN concentration, XG concentration, and primary fraction (PF) on the encapsulation efficiency (EE) and droplet size (DS) were systematically evaluated using response surface methodology (RSM) based on a Box–Behnken design (BBD). Microscopic and rheological analyses confirmed the formation of a rigid interfacial film around the droplets, leading to improved emulsion stability over one month of storage at 4, 25, and 40 °C. The release kinetics of chlortetracycline (CTC), used as a model drug, followed a Fickian diffusion mechanism, indicating efficient control of the release rate by the NaCN/XG interfacial complex. The optimized formulation (NaCN = 0.652%, XG = 0.339%, PF = 10%) yielded an encapsulation efficiency of 87.7% and a mean droplet size of 24.83 µm, demonstrating excellent predictive accuracy of the statistical model. The results highlight the potential of NaCN/XG complexes to produce acid-stable, biopolymer-based double emulsions capable of sustained release of bioactive compounds, making this system promising for food and pharmaceutical delivery applications. 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

The normal metabolism of transient starch in leaves plays a vital role in determining photosynthesis and final crop yield. However, the molecular mechanisms linking abnormal transient starch metabolism to premature leaf senescence remain unclear. Here, we isolate a rice mutant, lses1, with leaf yellowing and premature senescence, as well as excessive accumulation of starch granules in chloroplasts. Genetic analysis revealed that this trait is controlled by a single recessive nuclear gene. Through BSA-seq preliminary gene mapping, map-based cloning, and sequencing alignment, the candidate gene was pinpointed to LOC_Os02g40860 on chromosome 2, which encodes OsCKI1, a casein kinase I family member. The identity of LSES1 was confirmed functionally: genetic complementation with the native genomic sequence rescued the wild-type phenotype, while CRISPR/Cas9 knockout of the gene in wild-type plants recapitulated the premature senescence. This confirmed that LSES1/OsCKI1 is involved in the regulation of leaf senescence. Notably, one improved knockout line, KO-2, displayed significant agronomic improvements in grain length, grain width, number of productive ears, and number of filled grains per panicle, along with a significant increase in grain yield per plant, highlighting its potential breeding value. Subcellular localization and tissue-specific expression analysis showed that LSES1 is primarily nuclear-localized and constitutively expressed. Full article