Search Results (134)

This work evaluated how ferulic acid (FA) impacted the conformational properties of whey protein isolate (WPI) and altered the environmental tolerance of oleogel/water (Og/W) emulsions formulated with Ganoderma lucidum spore oil (GLSO). Molecular dynamics simulation analyses revealed that FA interacted with α-lactalbumin via hydrogen bonding and hydrophobic interactions, whereas it bound into the hydrophobic cavity of β-lactoglobulin through a “lock-and-key” mode driven primarily by hydrophobic forces. Fourier transform infrared spectroscopy analysis verified that such non-covalent forces triggered the dissociation and structural extension of WPI, which was manifested as a significant loss of α-helix and β-sheet architectures along with a corresponding rise in random coils. FA addition increased the positive charge, mean droplet size, interfacial contact angle and antioxidant stability of WPI-FA nanoparticles when the WPI-to-FA ratio exceeded 1:2 (i.e., WPI was in excess relative to FA). The GLSO-based Og/W emulsions exhibited a weak gel structure with predominantly elastic characteristics. Furthermore, WPI-FA nanoparticles fabricated at a 2:1 ratio minimized emulsion droplet size and imparted optimal stability to the Og/W emulsions, demonstrating superior freeze–thaw and salt resistance, alongside suppressed GLSO flavor release. This work provides critical insights into tailoring protein-polyphenol interactions to stabilize GLSO-based Og/W emulsion delivery systems for food applications. Full article

Cow’s milk allergy (CMA) is the most common food allergy in infants. Goat milk exhibits relatively low allergenicity and is widely regarded as a potential substitute for cow milk. However, the high sequence homology between cow and goat milk proteins may trigger cross-reactivity, significantly restricting the practical application of goat milk. This study aims to disrupt cross-linear epitopes of cow and goat milk proteins through enzymatic hydrolysis. We prepare hypoallergenic goat milk protein hydrolysates and systematically evaluate their desensitization effects. Bioinformatics methods were employed to predict the B-cell linear epitopes of six major allergens (α_S1-casein, α_S2-casein, β-casein, κ-casein, α-lactalbumin, and β-lactoglobulin) from cow, goat, and sheep milk, and sequence homology was analyzed through protein-protein Basic Local Alignment Search Tool (BLASTP). The results showed that the sequence similarity of homologous allergens among the three milk sources all exceeded 30%. Subsequently, six proteases (protamex, alcalase, pepsin, trypsin, papain and bromelain) were used to hydrolyze whole goat milk protein. Indirect enzyme-linked immunosorbent assay (ELISA) results indicated that all six hydrolysates significantly reduced immunoreactivity with antibodies against the five major cow’s milk allergens. Among them, alcalase exhibited significant efficacy against all five allergens; papain and protamex were particularly effective against β-casein; trypsin showed pronounced efficacy against α-lactalbumin and β-lactoglobulin; and bromelain and pepsin also significantly reduced immunoreactivity against certain allergens. Mass spectrometry revealed the peptide composition and epitope coverage of the hydrolysates: bromelain yielded two overlapping peptides (FAWPQY, LKDLKDY), protamex one (LAMAAS), Alcalase three (PPQSVLS, IPIQYVLS, LPYPYY), trypsin two (YLGYLEQL, AMAASDISLL), papain three (NEINQFYQK, FQSEEQQQTEDELQDK, AMAASDISL); and no matching peptide was detected for pepsin. These results confirmed at the molecular level that enzymatic hydrolysis can effectively disrupt cross-reactive epitopes. Full article

Background: Oral food challenge (OFC) remains the gold standard for diagnosing food allergy; however, it is time-consuming and associated with the risk of allergic reactions. The identification of reliable biomarkers capable of predicting OFC outcomes could improve patient selection and reduce the number of OFCs required in clinical practice. Among the most widely used biomarkers are skin prick tests (SPTs) and serum-specific IgE (sIgE) measurements, including component-resolved diagnostics. However, their diagnostic utility, as well as the comparability of different diagnostic platforms, remains incompletely defined in cow’s milk (CM) and hen’s egg (HE) allergy. Methods: We retrospectively analyzed results of diagnostic tests obtained in children undergoing OFCs to baked milk, raw milk, baked egg, and boiled egg. Diagnostic performance of SPTs to either allergen extracts or native food allergens, as well as sIgE measured using ImmunoCAP and ALEX2 platforms, was evaluated using receiver operating characteristic (ROC) analysis and logistic regression models. Results: Diagnostic performance varied across allergy phenotypes. SPTs to native food allergens demonstrated good discriminative capacity for raw milk and boiled egg allergy, whereas SPTs to standardized extracts showed limited diagnostic utility overall. In baked milk allergy, casein-sIgE measured by ImmunoCAP demonstrated the best diagnostic performance. In raw milk allergy, α-lactalbumin-sIgE measured using ALEX2 platform and β-lactoglobulin-sIgE measured by ImmunoCAP showed greater diagnostic relevance than casein-sIgE. In boiled egg allergy, SPT to raw egg yolk demonstrated the highest diagnostic value. In baked egg allergy, ovalbumin-sIgE measured using the ALEX2 platform showed the best performance, while ovomucoid-sIgE did not demonstrate clear superiority. ALEX2 showed good concordance with ImmunoCAP despite systematic differences in absolute sIgE values. Combined multivariable models provided only modest improvements over single predictors and did not achieve sufficient accuracy to replace OFCs. Conclusions: SPTs to either allergen extracts or native food allergens, as well as sIgE measurements, demonstrate moderate and heterogeneous utility in predicting OFC outcomes for CM and HE allergy. Although ALEX2 and ImmunoCAP showed comparable overall performance, platform-specific interpretation may be required. OFC remains indispensable for definitive diagnosis. Full article

In view of the importance of texture and colour characteristics for consumer acceptance of fermented dairy products, this study aimed to evaluate the associations between selected whey proteins, fat-soluble vitamins, and instrumental quality traits of organic plain yoghurts. Physicochemical analyses included acidity, total protein, fat, whey proteins (β-lactoglobulin, α-lactalbumin, bovine serum albumin, lactoferrin, and lysozyme), and fat-soluble compounds (vitamins A, D₃, and E, and β-carotene). Selected instrumental texture and colour attributes were also determined. Whey protein fractions were associated mainly with texture characteristics, whereas lipophilic vitamins and β-carotene were more closely related to colour attributes. Total protein content was positively associated with firmness (r = 0.510, p ≤ 0.05), while β-lactoglobulin was related to apparent viscosity (r = 0.705, p ≤ 0.05). In contrast, vitamin A, vitamin D₃, β-carotene, and fat content were negatively associated with lightness and whiteness, but positively related to yellowness and chroma. Lactoferrin and lysozyme showed stronger relationships with selected secondary texture attributes, particularly gumminess and cohesiveness. Multiple regression analysis indicated that firmness and apparent viscosity were only moderately explained by the selected compositional predictors, whereas colour attributes were more strongly associated with fat-fraction components. The remaining variability likely reflected additional microstructural and processing-related effects. Although the results should be interpreted as associations rather than direct causal relationships, the findings support the concept of protein-driven texture and fat-driven colour development in plain yoghurts and improve understanding of the relationships between bioactive milk components and instrumental plain yoghurt quality. Full article

The gut microbiota (GM) has become a key mediator of host health, with dietary manipulations promising ways of modulating the microbiome. This review focuses on the role of dairy bioactive (DB) compounds as precision modulators of intestinal microecology, including the whey proteins (WPs), including lactoferrin (LF), α-lactalbumin (LA), β-lactoglobulin, lysozyme (LZ), lactoperoxidase, glycomacropeptide (GMP), milk oligosaccharides (MOs), and bioactive peptides (BPs). This review compiles the existing evidence illustrating their dual-action mechanism through direct prebiotic activity and the promotion of beneficial taxa (Bifidobacterium, Lactobacillus, Faecalibacterium), along with selective antimicrobial activity and pathogen suppression. These compounds improve intestinal barrier integrity through tight junction (TJ) protein regulation, regulating short-chain fatty acid production, and modulating immune signaling pathways. Clinical evidence shows significant benefits in metabolism and inflammation among various populations. However, individual responses vary according to host factors such as enterotypes, FUT2 genotype, and baseline microbiota composition, suggesting the need for personalized intervention strategies. This review addresses critical knowledge gaps in dose–response relationships, long-term efficacy, and mechanistic pathways and suggests future directions for precision nutrition. By modifying molecular mechanisms in clinical applications, we have identified DB compounds as promising candidates for targeted modulation of the microbiota to optimize health and disease management. The review also brings together molecular mechanistic and clinically implementable, personalized dietary strategies, which have not been fully captured by previous reviews. It pinpoints gaps in knowledge related to dose–response characterization, long-term trial design, and multi-omics stratification that collectively define a new precision nutrition framework. In this approach, dairy-based intervention is planned for each person based on their microbial, genetic, and metabolic characteristics. Full article

This review aims to examine the immunological, anti-inflammatory, antiviral, and antibacterial activities of key whey and milk proteins, specifically lactoferrin, glycomacropeptide, β-lactoglobulin, α-lactalbumin and their derived peptides, particularly lactoferricin and lactoferrampin, highlighting their potential as preventive or therapeutic agents. Whey and dairy products represent complex biological matrices that, beyond their high nutritional value, serve as reservoirs of bioactive proteins and peptides with documented health-promoting properties. It has been reported that certain whey proteins (WPs) and whey-derived peptides may contribute to improvements in both innate and adaptive immunity, exert direct antiviral and antibacterial effects while also modulating host defenses through immunoregulatory, antioxidant, and anti-inflammatory activities. These mechanisms contribute not only to enhanced resistance against viral pathogens but also to maintaining intestinal homeostasis and microbiota balance, both of which are critical during infection. In recent years, particularly in the context of the COVID-19 pandemic, natural bioactive compounds derived from whey, and, more broadly, milk, have attracted increasing attention as potential adjuncts or alternatives to conventional antivirals, with reported activity not only against SARS-CoV-2, influenza but also other viral and microbial infections. Despite encouraging in vitro and in vivo evidence, clinical validation remains limited, and the antiviral and immunomodulatory effects of WPs still require deeper mechanistic clarification. Future research should focus on identifying molecular targets, as well as characterizing the pharmacokinetics and safety profiles of WPs and WP peptides across diverse clinical settings. At the same time, attention should be given to optimizing their application as nutraceuticals or functional dairy ingredients. Full article

Background/Objectives: Gastroesophageal reflux disease (GERD) is prevalent in infants and frequently managed with acid-suppressive medications that elevate gastric pH. This pilot study aimed to evaluate how varying gastric pH levels (2.5, 4.0 and 6.0) influence the hydrolysis of milk proteins in human milk (HM), cow’s milk-based infant formula (CMF), and goat milk-based infant formula (GMF). Methods: Samples were subjected to a 30 min in vitro gastric digestion using pediatric human gastric juice obtained from clinical donors. Protein degradation was analyzed via SDS-PAGE densitometry, comparing digested aliquots to undigested controls. Results: At pH 2.5, caseins were highly digested in all samples, especially in HM and GMF. At pH 4.0, GMF displayed an apparent 51% greater casein degradation relative to CMF and HM in this pilot analysis. α-lactalbumin degradation was markedly higher in GMF at all pH levels; notably, at pH 4.0 and 6.0, only GMF exhibited digestion of this protein. Albumin showed almost complete degradation in HM and GMF at pH 2.5, and GMF maintained greater degradation at higher pH levels. β-lactoglobulin (absent in HM) was better digested in GMF at pH 2.5, whereas CMF showed higher hydrolysis observed at pH 4.0 and 6.0. Lactoferrin digestion was most efficient in HM and GMF at pH 2.5, with no differences observed at higher pH levels. Conclusions: These preliminary findings suggest that GMF may offer digestive advantages for infants with GERD under pharmacological acid suppression, particularly regarding casein and α-lactalbumin breakdown at higher pH. The distinct digestion kinetics of CMF and GMF at different pH levels provide a physiological basis for targeted infant feeding strategies. Further large-scale studies are required to validate these exploratory observations. Full article

Balancing microbial safety and the retention of heat-sensitive components has long been a key issue in dairy processing research. This study systematically compared the effects of instantaneous ultra-high-temperature treatment (INF, 145–155 °C/0.09 s) with that of conventional pasteurization (75–95 °C/15 s) as well as ultra-high-temperature treatment (UHT, 135 °C/5 s), on the microbial evaluation, nutritional composition, and physicochemical quality of bovine milk. The results showed that all heat treatments completely inactivated Staphylococcus aureus, coliforms, while only UHT and INF achieved full spore elimination. In the INF group, α-lactalbumin remained almost completely native and native β-lactoglobulin retention was approximately 83% relative to raw milk. The retention of lactoferrin and immunoglobulin G was about 30% and 12% after INF treatment, respectively, which were higher than that of 13% and 8% in the 85 °C/15 s group, and complete denaturation in the 95 °C/15 s and UHT groups. Furthermore, vitamin B₂ remained stable after INF treatment. The glycation content of proteins was lower in INF treatment compared to conventional heat treatments, especially for the concentration of furosine, which was about 6–7 mg/100 g protein in the INF group, and 15 mg/100 g protein in the 95 °C/15 s group, and 67 mg/100 g protein in the UHT group. Overall, the INF process achieved sterilization equivalent to UHT while substantially reducing thermal load, thus better balancing microbial safety, nutritional integrity, and immune-active proteins, which provides a scientific basis for establishing standardized INF parameters and promoting high-quality dairy production. Full article

Breast cancer is the most common and deadliest cancer among women. While overexpression of specific markers guides disease stratification and has enabled the development of targeted therapies, identifying new therapeutic targets remains critical, particularly for aggressive subtypes lacking effective treatments. This study evaluated the expression of α-Lactalbumin (LALBA) and nucleolin (NCL) in breast cancer tissues from Mexican patients using gene expression analysis and immunohistochemistry. LALBA, a major milk protein normally expressed only during late pregnancy and lactation, was detected in nearly all tumor samples and showed higher levels in aggressive subtypes, with overexpression displaying a slight trend toward poorer overall survival. NCL, a multifunctional nucleolar protein, exhibited predominantly nuclear localization, with moderate expression associated with improved survival. Both proteins correlated with tumor immune features, including increased tumor-infiltrating lymphocytes (TILs) and PD-L1 expression for LALBA, and elevated CD8⁺ T cells, PD-L1, and TIM-3 expression for NCL. Overall, these findings suggest that LALBA and NCL are associated with tumor aggressiveness, immune context, and survival trends in breast cancer. Additional studies in larger cohorts are needed to define their clinical relevance. Full article

During gastrointestinal digestion, dietary proteins are hydrolyzed into peptides and free amino acids that modulate enteroendocrine function and satiety-related hormone secretion along the gut–brain axis, thereby contributing to obesity prevention. We investigated whey protein concentrate (WPC) as a source of bioactive peptides and evaluated the effects of its digests on cholecystokinin (CCK) secretion in STC-1 enteroendocrine cells by integrating the standardized INFOGEST in vitro digestion protocol, peptidomics (LC–MS/MS), and in silico bioactivity prediction. In STC-1 cells, the <3 kDa intestinal peptide fraction exhibited the strongest CCK stimulation, positioning these low-molecular-weight peptides as promising bioactive components for satiety modulation and metabolic health applications. Peptidomic analysis of this fraction identified short sequences derived primarily from β-lactoglobulin (β-La) and α-lactalbumin (α-La), enriched in hydrophobic and aromatic residues, including neuropeptide-like sequences containing the Glu–Asn–Ser–Ala–Glu–Pro–Glu (ENSAEPE) motif of β-La f(108–114). In silico bioactivity profiling with MultiPep predicted antihypertensive, angiotensin-converting enzyme (ACE)–inhibitory, antidiabetic, dipeptidyl peptidase-IV (DPP-IV)–inhibitory, antioxidant, antibacterial, and neuropeptide-like activities. Overall, digestion of WPC released low-molecular-weight peptides and amino acids that enhanced CCK secretion in vitro; these findings support their potential use in nutritional strategies to enhance satiety, modulate appetite and energy intake, and improving cardiometabolic health. Full article

Cheese whey, the major by-product of the dairy industry, poses an environmental challenge due to its high organic load but simultaneously represents a nutrient-dense matrix suitable for biotechnological valorization. This review synthesizes recent advances positioning whey as (i) a fermentation substrate for lactic acid bacteria, yeasts/fungi, and microalgae, enabling the production of functional biomass, organic acids, bioethanol, exopolysaccharides, enzymes, and wastewater bioremediation; (ii) a platform for postbiotic generation, supporting cell-free preparations with functional activities; and (iii) a food-grade encapsulating material, particularly through whey proteins (β-lactoglobulin, α-lactalbumin), which can form emulsions, gels, and films that protect biotics and bioactive compounds during processing, storage, and gastrointestinal transit. We analyze key operational variables (whey type and pretreatment, supplementation strategies, batch and continuous cultivation modes), encapsulation routes (spray drying, freeze-drying, and hybrid protein–polysaccharide systems), and performance trade-offs relevant to industrial scale-up. Finally, we outline future directions, including precision fermentation, mixed-culture processes with in situ lactase activity, microfluidics-enabled encapsulation, and life-cycle assessment, to integrate product yields with environmental performance. Collectively, these strategies reframe whey from a high-impact waste into a circular bioeconomy resource for the food, nutraceutical, and environmental sectors. Full article

Donkey milk is increasingly recognized as a functional food due to its unique nutritional profile and richness in bioactive compounds. This longitudinal observational study investigated changes in both chemical composition (total solids, protein, fat, lactose, and ash) and immune-active proteins (lactoferrin, α-lactalbumin, β-lactoglobulin, and lysozyme) across lactation. A total of 153 donkey milk samples were collected from five farms from very early (1–3 days in milk) to late lactation (30–210 days in milk). Chemical composition was determined using mid-infrared spectroscopy, while the concentrations of the immune-active proteins were determined by ELISA Quantitative Sandwich. Chemical analysis showed high values in colostrum, including total solids (10.13%), protein (3.1%), and ash (0.73%), which declined progressively during lactation to 8.45%, 1.14%, and 0.64%, respectively. Fat varied modestly, between 0.55 and 0.25%, while lactose remained stable at 5.75–6.41%. In parallel, bioactive proteins measured between 31 and 210 days exhibited distinct trajectories. Lactoferrin increased from 0.07 to 0.14 mg/mL, α-lactalbumin peaked mid-lactation at 2.58 mg/mL (compared with 1.91 mg/mL early and 2.25 mg/mL late), β-lactoglobulin declined from 0.84 to 0.55 mg/mL, and lysozyme decreased from 0.95 mg/mL early to 0.64 mg/mL late. Across lactation, we observed dilution of total solids and protein, relatively stable lactose and fat, and distinct trajectories of lactoferrin, α-lactalbumin, β-lactoglobulin, and lysozyme, indicating that donkey milk modulates rather than loses its protective protein profile. These results refine reference values for donkey milk and support its nutraceutical relevance for human nutrition and health. Full article

Background: The study aimed to evaluate the efficacy of a combination of Dioscorea villosa (containing diosgenin), vitamin D, and α-lactalbumin, in women with Polycystic Ovary Syndrome (PCOS) phenotype D. The primary objective was to investigate improvements in menstrual cycle regularity. Methods: A total of 24 women aged 22–34 years with PCOS phenotype D received daily supplementation with 600 mg Dioscorea villosa (120 mg diosgenin), 100 mg α-lactalbumin, and 50 μg vitamin D for six months. Clinical and biochemical assessments, including hormonal profiling and menstrual cycle monitoring, were conducted at baseline (T0), after 3 months (T1), and after 6 months (T2). Results: The treatment led to a statistically significant improvement in menstrual cycle regularity: eumenorrhea was achieved in 50% of patients at T2, compared to 0% at baseline. Significant changes were also observed in luteinizing hormone (LH), follicle-stimulating hormone (FSH), and the LH/FSH ratio, alongside a reduction in insulin and HOMA-index at T1. No adverse events were reported. Conclusions: The combination of Dioscorea villosa, vitamin D, and α-lactalbumin promotes menstrual cycle regularization in women with PCOS phenotype D. The positive result suggests a beneficial role of the treatment when administered to this specific subtype of PCOS and supports the use of targeted nutraceutical therapy as an alternative to conventional treatments, especially in non-hyperandrogenic PCOS patients. Full article

Bioactive peptides (BAPs) from dairy products have garnered increasing attention as natural agents with health-promoting properties, including antihypertensive, antioxidant, antimicrobial, immunomodulatory, opioid, and antidiabetic activities. This systematic review synthesizes research published between 2014 and 2024, retrieved from Scopus and PubMed, and selected according to PRISMA guidelines. A total of 192 studies met the inclusion criteria, collectively reporting over 3200 distinct peptides, with antihypertensive sequences, predominantly angiotensin-converting enzyme (ACE) inhibitors, constituting the largest category (n = 1237). β-casein was the principal precursor across bioactivities, followed by αs1-casein, β-lactoglobulin, and α-lactalbumin. Peptides were primarily produced via enzymatic hydrolysis, microbial fermentation, and gastrointestinal digestion, with peptide profiles influenced by the type of milk, microbial strains, and processing conditions. While cow’s milk remained the dominant source, investigations into goat, sheep, camel, buffalo, and donkey milk revealed species-specific biopeptides. Recent advances in proteomics have enhanced peptide identification and bioactivity prediction, enabling the discovery of novel sequences. These findings underscore the significant potential of dairy-derived BAPs as functional food components and nutraceutical ingredients, while highlighting the need for further in vivo validation, bioavailability studies, and broader exploration of underrepresented milk sources. Full article

The functional properties of proteins are closely related to their structure and conformation. The effects of glycosylation and pH on the structural and conformational changes in whey protein isolate (WPI) were investigated using multispectral technology. More and higher-molecular-weight molecules of WPI–dextran conjugates (WDCs) with increased degrees of glycosylation (DGs) in SDS-PAGE occurred at the expense of band intensities of α-lactalbumin, β-lactoglobulin, and bovine serum albumin. The higher wavenumber shift in FTIR peaks of WPI after glycosylation in the Amide I, II, and III regions and the decrease in its intensity occurred. The maximum absorption wavelength (λ_max) of UV-Vis spectra of WPI before and after glycosylation in the range of 260–290 nm showed no significant difference in a pH range of 2.0–10.0. Moreover, the UV-Vis absorption intensities of WDCs at λ_max around 278 nm were highly and positively correlated with their DGs. The λ_max and intensities of total intrinsic fluorescence spectra of Tyr and Trp residues in WDCs with an increase in DGs had an obvious redshift and decrease, respectively. Although the intensities of synchronous fluorescence spectra of individual Tyr or Trp residues in WDCs with an increase in DGs also gradually decreased, the λ_max of the former and latter had a blueshift and redshift, respectively. UV-Vis absorption and fluorescence spectroscopies indicated that the changes in the λ_max and intensity of WPI were closely related to the protonation states of carbonyl groups and free amino groups and the degree of glycosylation. This work may be beneficial for understanding the structural and conformational changes in proteins by measuring the microenvironment around Tyr and/or Trp residues in proteins using UV-Vis absorption and synchronous fluorescence spectroscopies, providing a promising technique for quantitatively monitoring the degree of glycosylation (DG) in a rapid and practical way without any chemical reagents using UV-Vis absorption spectroscopy. Full article