Search Results (195)

Milk is a nutritionally rich food and a frequent target of economically motivated adulteration, particularly through substitution with lower-cost milk types. Over the past decade, significant progress has been made in the authentication of milk using advanced proteomic and chemometric approaches, with a focus on the discovery and application of protein and peptide biomarkers for species differentiation and fraud detection. Recent innovations in both top-down and bottom-up proteomics have markedly improved the sensitivity and specificity of detecting key molecular targets, including caseins and whey proteins. Peptide-based methods are especially valuable in processed dairy products due to their thermal stability and resilience to harsh treatment, although their species specificity may be limited when sequences are conserved across related species. Robust chemometric approaches are increasingly integrated with proteomic pipelines to handle high-dimensional datasets and enhance classification performance. Multivariate techniques, such as principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), are frequently employed to extract discriminatory features and model adulteration scenarios. Despite these advances, key challenges persist, including the lack of standardized protocols, variability in sample preparation, and the need for broader validation across breeds, geographies, and production systems. Future progress will depend on the convergence of high-resolution proteomics with multi-omics integration, structured data fusion, and machine learning frameworks, enabling scalable, specific, and robust solutions for milk authentication in increasingly complex food systems. Full article

Crustins are a family of cysteine-rich antimicrobial peptides (AMPs), predominantly found in crustaceans, and play important roles in innate immunity. However, among the many reported crustins, few studies have explored their immunomodulatory functions. In this study, we investigated the immune function of a type I crustin (LvCrustinIa-2) in Litopenaeus vannamei, with particular emphasis on comparing the roles of its different domains. LvCrustinIa-2 possesses cationic patchy surface and amphipathic structure, and its expression was significantly induced in hemocytes after pathogen challenge. Both the recombinant LvCrustinIa-2 (rLvCrustinIa-2) and its whey acidic protein (WAP) domain (rLvCrustinIa-2-WAP) exhibited significant inhibitory activities against the tested Gram-positive bacteria. They also showed binding affinity not only for Gram-positive bacteria but also for Gram-negative bacteria. Furthermore, rLvCrustinIa-2 induced membrane leakage and structure damage in the target bacteria. Notably, chemotaxis assays revealed that rLvCrustinIa-2 and the synthetic cysteine-rich region (LvCrustinIa-2-CR) significantly enhanced the chemotactic activity of shrimp hemocytes in vitro. Knockdown of LvCrustinIa-2 triggered significant transcriptional activation of genes involved in calcium transport, inflammation, redox regulation, and NF-κB pathway. Taken together, these findings elucidate the distinct roles of the cysteine-rich region and WAP domain in type Ia crustin and provide the first evidence of a crustacean AMP with chemotactic and immunomodulatory activities. Full article

Tofu quality is critically influenced by coagulants, though their impact on protein digestibility remains underexplored. This study aimed to investigate the effects of calcium sulfate (CaSO₄), magnesium chloride (MgCl₂), and their combination (CaSO₄ + MgCl₂) on the physicochemical properties and protein digestibility of tofu. Water-holding capacity, cooking loss, texture, protein composition, and protein digestibility were analyzed. The results showed that the CaSO₄ + MgCl₂ combination yielded a water-holding capacity of 99.16%, significantly higher than CaSO₄ tofu (93.73%) and MgCl₂ tofu (96.82%), while reducing cooking loss to 2.03% and yielding the highest hardness (897.27 g) and gumminess (765.72). Electrophoresis revealed distinct protein retention patterns, with MgCl₂ (0.6% w/v) forming denser gels that minimized protein leakage into soy whey. During in vitro digestion, MgCl₂-coagulated tofu exhibited superior soluble protein release (5.33 mg/mL after gastric digestion) and higher intestinal peptide (5.89 mg/mL) and total amino acid (123.06 μmol/mL) levels, indicating enhanced digestibility. Conversely, the CaSO₄ + MgCl₂ combination showed delayed proteolysis in electrophoresis analysis. These findings demonstrate that coagulant selection directly modulates tofu’s texture, water retention, and protein bioavailability, with MgCl₂ favoring digestibility and the hybrid coagulant optimizing physical properties. This provides strategic insights for developing nutritionally enhanced tofu products. Full article

Background/Objectives: Hepatic steatosis, characterized by abnormal fat accumulation in the liver, is a major health concern with limited effective treatments. Goat milk whey proteins have demonstrated various therapeutic benefits. This study aimed to evaluate the hepatoprotective effects of goat whey protein hydrolysate (GWPH) on high-fructose corn syrup (HFCS)-induced hepatic steatosis in a murine model. Methods: The GWPH was prepared through enzymatic hydrolysis using Alcalase^® and divided into fractions: GWPH03 (<3 kDa), GWPH0310 (3–10 kDa), GWPH1030 (10–30 kDa), and GWPH30 (>30 kDa). These fractions were administered to respective GWPH treatment groups at 200 mg/kg b.w/day via intragastric gavage for 8 weeks, with HFCS provided to all groups except the Naïve group. After dietary intervention, an oral glucose tolerance test (OGTT) was performed, and the mice were then sacrificed for further analysis. Results: Our results demonstrate that GWPH mitigates HFCS-induced hepatic steatosis, reduces body weight gain, improves glucose homeostasis, alleviates liver injury, and regulates hepatic lipid metabolism. Notably, GWPH treatment significantly suppressed hepatic fatty acid synthase (FASN) expressions, indicating reduced de novo lipogenesis (DNL). Molecular docking of the identified peptides from GWPH—particularly PFNVYNVV, which showed strong binding affinity for KHK—suggests that it has potential as a competitive inhibitor of fructose metabolism. Conclusions: Collectively, our findings suggest that GWPH and its derived peptides could be promising candidates for managing hepatic steatosis and related metabolic abnormalities. Full article

The whey protein (WP) fraction represents 18–20% of the total milk nitrogen content. It was originally considered a dairy industry waste, but upon its chemical characterization, it was found to be a precious source of bioactive components, growing in popularity as nutritional and functional food ingredients. This has generated a remarkable increase in interest in applications in the different sectors of nutrition, food industry, and pharmaceutics. WPs comprise immunoglobulins and proteins rich in branched and essential amino acids, and peptides endowed with several biological activities (antimicrobial, antihypertensive, antithrombotic, anticancer, antioxidant, opioid, immunomodulatory, and gut microbiota regulation) and technological properties (gelling, water binding, emulsification, and foaming ability). Currently, various process technologies and biotechnological methods are available to recover WPs and convert them into BioActive Peptides (BAPs) for commercial use. Additionally, in silico approaches could have a significant impact on the development of novel foods and/or ingredients and therapeutic agents. This review provides an overview of current and emerging methods for the production, selection, and application of whey peptides, offering insights into bioactivity profiling and potential therapeutic targets. Recent updates in legislation related to commercialized WPs-based products are also presented. Full article

Cosmetology is one of the fastest-growing scientific areas, and within it, individual needs and preferences have to be considered. Specifically, cosmetic products with incorporated biological macromolecules, i.e., proteins and peptides, that contribute to improved skin features are gaining in importance. Similar to other fields, cosmetology is also faced with the zero-waste paradigm and strives for a collaboration with other industries. Whey is a co-product in milk production and represents a high environmental burden. In this regard, the idea of the present study was to utilise whey in order to develop sustainable cosmetic products, i.e., cleansing hydrogel and shampoo. The initial phase of the study was dedicated to the development of an optimised hydrogel and shampoo base, followed by whey integration and an in-depth physico-chemical characterisation of both prototypes. In the subsequent phases, particular emphasis was placed on evaluating the potential skin irritancy of the whey-based formulations in vitro, complemented by in vivo assessment on volunteers. The results obtained indicate that the incorporation of whey at concentrations of up to 30% (m/m) is feasible for both formulation types. Moreover, neither product exhibited any irritative effects and a study on volunteers showed that whey has great potential in terms of providing adequate skin hydration. Taken together, all the findings support the development of advanced cosmetic formulations with a zero-waste concept built-in, thus offering a promising platform for cross-sector collaboration, and representing a meaningful step toward potential hydrogel and shampoo commercialisation. Full article

The aim of this work is to highlight the beneficial effects of bioactive peptides present in fermented whey (FW) and carotenoids from pumpkin (P) against the pro-oxidant effects of aflatoxin B1 and ochratoxin A at the neuronal level. For this purpose, SH-SY5Y human neuroblastoma differentiated cells were exposed to (A) mycotoxins, (B) the digesta of mycotoxin-contaminated bread formulated with P, or (C) bread enriched with FW + P. A proteomic approach using HPLC-MS/MS-QTOF was then employed to characterize the metabolic pathways affected by the presence of these components, as well as their ability to modulate the toxic effects exacerbated by mycotoxins. Gene ontology functional analysis revealed proteins primarily associated with nucleosome structure, such as the H3-H4 tetramer, H2A-H2B dimer, and HIRA, which were overexpressed in the presence of mycotoxins and, interestingly, downregulated with the addition of the functional ingredients. Additionally, important metabolic pathways associated with the RHO GTPase family, estrogen-dependent gene expression, and androgen receptor transcription stimulated by PKN1 activation were discovered. Network interaction analysis highlighted the modulation of cytoskeletal dynamics, cell migration, and stress responses. These findings provide novel insights into the neuroprotective potential of functional food components, supporting their use in mitigating mycotoxin-induced neuronal damage and opening new avenues for dietary-based neuroprotection strategies. Full article

This study investigates the potential of blackberry juice fortified with whey as a carrier for transporting and protecting lactic acid bacteria (LAB). The interactions between whey proteins and the juice were examined to assess their impact on probiotic stability and protection during storage and passage through the gastrointestinal tract. Additionally, the study explored how this combination influences the antioxidant properties of the product. The results indicated that the blackberry juice and whey protein mixture provided moderate protection to Lacticaseibacillus rhamnosus GG compared to the positive control (inulin), suggesting that whey proteins may enhance probiotic viability. Proteolytic analysis revealed progressive protein hydrolysis during fermentation, leading to the release of bioactive peptides, indicating the formation of compounds with potential functional benefits. Moreover, samples inoculated with LAB exhibited higher antioxidant activity than those without inoculum. This research demonstrates the promise of fermented blackberry juice fortified with whey proteins as an effective probiotic delivery system. It opens new possibilities for developing functional foods to promote intestinal health and overall well-being. Full article

Cow milk allergy (CMA) is prevalently observed among infants and young children, exerting adverse effects on their growth and quality of life. Oral immune tolerance (OIT) is a more effective method for the prevention and treatment of CMA. The site of OIT is mainly in the gastrointestinal tract, so this article reviews the composition and structural characteristics of intestinal immune system, the molecular mechanisms of immune tolerance by regulatory T cells (Treg), dendritic cells, and gut microbiota. In addition, this paper summarizes the research progress of T cell epitope peptides of β-lactoglobulin and α-lactalbumin in whey protein hydrolysates. The mechanism of OIT induced by whey protein hydrolysate or whey protein combined with other anti-allergic components (phenolic compounds, probiotics, etc.) is overviewed to provide new ideas for the development of hypoallergenic infant formula. Full article

Campylobacter jejuni (C. jejuni) is the primary Campylobacter species and a major cause of foodborne illness associated with poultry products. This review focuses on lactic acid bacteria (LAB), especially Lactobacillus species, and acid whey as a dairy by-product for C. jejuni control in poultry as a sustainable method. LAB strains L. crispatus exhibit a cecal colonization reduction of >90% by competitive exclusion and bacteriocin activity, while L. johnsonii FI9785 decrease bacterial load 4–5 log₁₀. Acid whey, which is abundant in organic acids (e.g., lactic acid) and bioactive peptides (e.g., lactoferrin), reduces C. jejuni viability, decreasing the food product contamination on the carcass for a short time by 40%. LAB antimicrobial function becomes more effective when used with acid whey, although specific farm-related variables require additional optimization. Some of the key strategies include co-encapsulating LAB with acid whey or plant-derived antimicrobials for improving survival, conducting in vivo trials in commercial farm conditions to evaluate scalability, and adding whey into feed (1–2% inclusion) or applying it as a pre-slaughter spray. These strategies enable the antibiotic-free production and circular economy goals through repurposing low-cost acid whey. Future studies should directly compare them with standard antimicrobials to confirm their scalability for poultry safety. Full article

The valorization of cheese whey, a rich by-product of the dairy industry that is rich in lactose (approx. 70%), proteins (14%), and minerals (9%), represents a promising approach for microbial fermentation. With global whey production exceeding 200 million tons annually, the high biochemical oxygen demand underlines the important need for sustainable processing alternatives. This review explores the biotechnological potential of whey as a fermentation medium by examining its chemical composition, microbial interactions, and ability to support the synthesis of valuable metabolites. Functional microorganisms such as lactic acid bacteria (Lactobacillus helveticus, L. acidophilus), yeasts (Kluyveromyces marxianus), actinobacteria, and filamentous fungi (Aspergillus oryzae) have demonstrated the ability to efficiently convert whey into a wide range of bioactive compounds, including organic acids, exopolysaccharides (EPSs), bacteriocins, enzymes, and peptides. To enhance microbial growth and metabolite production, whey fermentation can be carried out using various techniques, including batch, fed-batch, continuous and immobilized cell fermentation, and membrane bioreactors. These bioprocessing methods improve substrate utilization and metabolite yields, contributing to the efficient utilization of whey. These bioactive compounds have diverse applications in food, pharmaceuticals, agriculture, and biofuels and strengthen the role of whey as a sustainable biotechnological resource. Patents and clinical studies confirm the diverse bioactivities of whey-derived metabolites and their industrial potential. Whey peptides provide antihypertensive, antioxidant, immunomodulatory, and antimicrobial benefits, while bacteriocins and EPSs act as natural preservatives in foods and pharmaceuticals. Also, organic acids such as lactic acid and propionic acid act as biopreservatives that improve food safety and provide health-promoting formulations. These results emphasize whey’s significant industrial relevance as a sustainable, cost-efficient substrate for the production of high-quality bioactive compounds in the food, pharmaceutical, agricultural, and bioenergy sectors. Full article

Bioactive peptides derived from milk proteins offer promising potential that can be unlocked through hydrolysis. Enzymatic hydrolysis is particularly noteworthy because of its mild conditions and its efficacy in producing peptides with various biological activities. This study focused on creating whey protein hydrolysates using three enzymes: pepsin, trypsin, and papain. The degree of hydrolysis and the antioxidant properties of the resulting peptides were evaluated, and papain demonstrated the highest degree of hydrolysis, leading to its selection for further investigation. LC-MS was employed to identify peptide sequences from the papain-derived hydrolysate, resulting in the identification of 107 distinct peptide sequences These peptides were predicted to exhibit a range of potential biological activities, including antihypertensive, antidiabetic, antioxidant, antimicrobial, and immunomodulatory effects, as well as roles in regulating glucose homeostasis, maintaining cardiovascular health, and supporting overall metabolic function. In vitro tests revealed the significant antioxidant and antibacterial properties of the hydrolysate, confirming the potential of papain-derived peptides for use in functional food and pharmaceutical applications. The novelty of this study lies in the identification of novel peptides with promising biological activities. Additional in vitro and in vivo studies are required to fully elucidate the health benefits of these peptides. Full article

This study aimed to evaluate the potential of lactic acid bacteria (LAB) isolated from organic acid whey as an alternative to nitrites in heat-treated organic sausages. Eleven LAB strains were screened for their ability to develop sensory characteristics similar to traditionally cured meat. Based on the results, Lactiplantibacillus plantarum S21 was selected for further experiments. Four sausage treatments were produced: control cured (C), salted (S), salted with L. plantarum S21 at 10⁷ CFU/g (LP), and salted with acid whey (AW). The pH value, oxidation-reduction potential (ORP), antioxidant activity of peptides (ABTS•+), thiobarbituric acid-reactive substance (TBARS), fatty acid profile, and microbiological quality were assessed post-production and after 14 days of cold storage. After production, the LP and AW sausages had a lower pH than the cured (C) and uncured (S) control samples. LP sausages exhibited a stable pink colour due to myoglobin conversion to nitrosylmyoglobin, comparable to the cured control. The LP sausages were similar in overall sensory quality to the cured (C) samples and were superior to the S and AW sausages after storage. The lowest ORP value was observed in treatment C after production, whereas after storage, no significant differences were found between the treatments. The highest antioxidant activity of peptides was observed in the LP sausages. It was shown that the LP and AW treatments had lower saturated fatty acid content and higher monounsaturated and polyunsaturated fatty acid content than the C and S treatments. Nevertheless, the C treatment had the lowest TBARS value. Lower total viable counts were found in the C and LP treatments than in the S and AW treatments after storage. Our research demonstrates the potential of L. plantarum S21 for producing heat-treated sausages without nitrites, assuming the implementation of additional anti-botulinum barriers. Nevertheless, further studies on the role of bacteria in meat oxidation processes are needed. Full article

Purpose: Heart failure (HF) impairs physical performance and increases the incidence of orthostatic hypotension (OH). Individuals with OH have a higher risk of falls, which are a major source of morbidity and mortality in older adults. Dietary protein supplementation can improve physical performance in healthy older adult individuals; however, its effect on OH in older adult patients with HF is unknown. Methods: Twenty-one older adult patients with mild-to-moderate HF were randomized to placebo or protein supplementation. Dietary protein was supplemented with whey protein so the total protein intake for each participant was 1.2 g/kg bodyweight/day, plus 1 g/day of the amino acid l-carnitine for 16 weeks. Susceptibility to OH was assessed using a head-up tilt test, blood markers, and a functional test (6 min walk) at baseline and 16 weeks. Results: There were no differences in tilt test responses or 6 min walk test (6MWT) distances. The protein-supplement group had a significant increase in 6MWT pulse pressures post-walk after 16 weeks of treatment as compared to placebo. However, the tachycardia observed at baseline after 6MWT in the protein group was not seen at the end of the study. There was also a trend towards lower levels of brain naturetic peptide (proBNP) in the protein group vs. placebo at 16 weeks. Conclusions: The improved pulse-pressure response to exertion and positive trends in proBNP in this pilot study suggest that dietary supplementation may improve cardiovascular function and general health in individuals with HF and that larger future studies are justifiable. Full article

Sweet whey (SW), a by-product of cheese production, has potential immunomodulatory properties that could be beneficial in preventing inflammation-related diseases. This study investigated the effects of SW derived from bovine, caprine, ovine, or an ovine/caprine mixture of milk on inflammation-related gene expression in THP-1-derived macrophages, both with and without LPS stimulation. Cells were treated with SW-D-P3 (a fraction smaller than 3 kDa produced by in vitro digestion), and the expression of inflammation-related genes was assessed using quantitative PCR. Results showed that the expression of TLR2 and ICAM1 was attenuated in non-LPS-stimulated macrophages treated with SW-D-P3, regardless of animal origin. Moreover, the expression of TLR4, IL1B, and IL6 was decreased and the expression of an NF-κB subunit RELA and CXCL8 was elevated in a subset of samples treated with SW-D-P3, depending on the milk source. In LPS-challenged cells, the expression of CXCL8 was upregulated and the expression of IRF5 and TNFRSF1A was downregulated in SW-D-P3-treated cells, regardless of animal origin. On the other hand, a number of inflammation-related genes were differentially expressed depending on the animal origin of the samples. Moreover, the higher IL10 expression observed in cells treated with ovine/caprine SW-D-P3 compared to those treated with SW-D-P3 of bovine, caprine, or ovine origin suggests an anti-inflammatory response, in which alternatively activated macrophages (M2 polarization phenotype) may participate. Overall, these findings suggest that incorporating SW into the food industry, either as a standalone ingredient or supplement, may help to prevent inflammation-related diseases. Full article