Search Results (394)

This study evaluated the survival of encapsulated and free probiotic strains (Lacticaseibacillus paracasei Shirota and Lacticaseibacillus rhamnosus GG) in orange juice during storage and in simulated gastrointestinal tract (GIT) conditions and their effect on the survival of Salmonella enterica. Samples were inoculated with free or encapsulated probiotics in whey protein isolate–gum Arabic matrix in 9.00 log CFU/mL population level and were stored at 4 °C and 12 °C for five days. Additionally, samples were co-inoculated with S. enterica 3-strain cocktail at 1.70 log CFU/mL. Samples were withdrawn daily, and microbiological analysis, pH, and sensory evaluation were conducted. Survival of probiotics and the pathogen were further assessed under GIT simulation conditions. Results demonstrated that both free and encapsulated probiotics maintained high population levels (9.00 log CFU/mL) during storage. During GIT simulation, free probiotic population reduced to 3.80 log CFU/mL, in contrast to the encapsulated cells that remained at 6.80–7.00 log CFU/mL after 2 h of the intestinal phase, confirming the protective role of microencapsulation. S. enterica population survived in control and when co-cultured with encapsulated probiotics until the end of storage in populations of 1.7 ± 0.06 log CFU/mL; however, it was reduced to 0.80 log CFU/mL when co-cultured with free probiotics. Salmonella survived during GIT simulation, in control samples, whereas the pathogen co-cultured with probiotics lead to complete S. enterica elimination. Notably, during the intestinal phase, the encapsulated probiotics effectively eliminated S. enterica, maintaining their viability in high population levels. These results highlight that encapsulating probiotics can improve both the functional and sensory characteristics of probiotic fruit juices while supporting high probiotic viability and thus suppression of pathogenic microorganisms in the intestinal environment. Full article

This study proposes a dual-approach strategy to formulate reduced-fat milk chocolate by combining cocoa butter (CB) substitution with emulsifier-based rheological optimization. CB was partially replaced at 20%, 30%, and 40% using whey protein isolate (WPI) and inulin (IN) blends (70:30, 50:50, 30:70 w/w). CB reduction increased plastic viscosity and yield stress, particularly in WPI-rich systems. The 50:50 WPI:IN ratio consistently minimized rheological drawbacks while maintaining melting, texture, and sensory quality. Caloric content was reduced by up to 9% (~50 kcal/100 g), most notably in IN-dominant samples. To overcome flow challenges at high substitution levels, emulsifiers—lecithin, ammonium phosphatide (AMP), and polyglycerol polyricinoleate (PGPR)—were assessed. AMP (≤0.5% w/w) and PGPR (0.15–0.3% w/w) effectively reduced viscosity and yield stress; lecithin showed limited effect above 0.6%. The optimized system (0.5% AMP + 0.15% PGPR) applied to 40% CB-reduced chocolate with 50:50 WPI:IN restored desirable rheology (3.42 Pa·s viscosity; 7.91 Pa yield stress) and improved mouthfeel and acceptability. This integrated formulation enables significant fat and calorie reduction without compromising product quality, supporting the development of healthier chocolate products. Full article

The interfacial and foam properties of proteins can be enhanced by altering the interactions between polyphenols and proteins. The aim of this study was to determine the influence of gallic acid (GA) on the structural properties of whey protein isolate (WPI), specifically focusing on particle size, potential, and surface hydrophobicity, as well as the subsequent alterations in its interfacial and foam properties when utilized as a foaming agent. An increase in turbidity and a decrease in particle size suggested the formation of a soluble complex between GA and WPI at a pH of 6. The results from fluorescence spectroscopy and surface hydrophobicity analyses indicated that the primary interactions between GA and WPI are characterized by hydrogen bonding and hydrophobic interactions. The reduction in particle size enhances the capacity of WPI/GA complexes to lower the surface pressure, thereby demonstrating significant efficacy at the macroscopic scale. Furthermore, the structural connectivity of GA facilitates the formation of a stable interfacial film at the air–water interface by WPI/GA, resulting in high foam stability at a macroscopic level. This research contributes to a deeper understanding of the application of protein–polyphenol complexes as surfactants and provides theoretical support for their use in food applications. Full article

Transglutaminase (TGase) improves protein structure by facilitating cross-linking reactions. However, the effects of TGase on the physicochemical properties of whey protein isolate (WPI)–soymilk complexes and their applications in yuba remain unclear. Therefore, the impacts of TGase concentration on the free sulfhydryl content, free amino content, particle size, and structure of WPI–soymilk complexes and their film-forming properties were studied. The results showed that the physicochemical properties of the composite soymilk were changed by the TGase-induced cross-linking reaction of protein. Compared with the composite soymilk without TGase modification, the particle size of the WPI–soymilk complexes increased from 707.99 ± 9.47 nm to 914.41 ± 2.8 nm as the TGase concentration increased, and the complexes remained relatively stable at low TGase concentrations. TGase modification changed the tertiary structure of the WPI–soymilk complexes. The composite yuba with 0.01% and 0.03% levels of TGase had a higher β-sheet content than composite yuba without addition of TGase. The surface hydrophobicity of composite soymilk was decreased by all the addition levels of TGase. Meanwhile, the TGase-modified composite protein with 0.03% TGase had the lowest free sulfhydryl (35.92 μg/g) and amino groups (0.46). Additionally, the tensile strength of the composite yuba with 0.05% TGase addition reached a peak of 1.66 ± 0.02 MPa, which was 7.8% higher than that of the composite yuba without TGase addition. The SEM results revealed that the composite yuba with 0.01–0.03% TGase addition exhibited a dense and non-porous film structure. Moreover, all the composite yuba with TGase addition had a reduced rate of yuba cooking loss. This study contributes to enhancing the yield and mechanical properties of traditional yuba. Full article

The aim of this work was to produce bone scaffolds containing whey protein isolate and pearl powder and to conduct a preliminary assessment of the biomedical potential in vitro and in vivo. This included analysis of structural, physicochemical, mechanical, and biological properties, which revealed that biomaterials containing pearl powder exhibited an enhanced porous structure, increasing absorptive properties, and decreasing proteolytic capacity with increasing inorganic component content. Pearl powder content in the biomaterials did not clearly influence their mechanical properties or their ability to release calcium ions, as well as proteins. Extracts obtained from all tested biomaterials showed no cytotoxicity in vitro. The surfaces of all biomaterials promoted normal human osteoblast growth, proliferation, and osteogenic differentiation. Furthermore, all biomaterials did not display toxicity in vivo, but no changes in Danio rerio were observed after evaluation of the biomaterial containing the highest amount of pearl powder–10% v/w (marked as WPI/P10). Taking all the obtained results into account, it appears that this biomaterial can be promising for bone scaffolds and similar applications, thanks to its porous structure, high cytocompatibility in vitro, and lack of toxicity in vivo. However, advanced studies will be conducted in the future. Full article

The rising demand for sustainable protein is driving interest in insects as a raw material for advanced food ingredients. This review collates and critically analyses over 300 studies on the conversion of crickets, mealworms, black soldier flies, and other farmed species into powders, protein isolates, oils, and chitosan-rich fibers with targeted techno-functional roles. This survey maps how thermal pre-treatments, blanch–dry–mill routes, enzymatic hydrolysis, and isoelectric solubilization–precipitation preserve or enhance the water- and oil-holding capacity, emulsification, foaming, and gelation, while also mitigating off-flavors, allergenicity, and microbial risks. A meta-analysis shows insect flours can absorb up to 3.2 g of water g⁻¹, stabilize oil-in-water emulsions for 14 days at 4 °C, and form gels with 180 kPa strength, outperforming or matching eggs, soy, or whey in specific applications. Case studies demonstrate a successful incorporation at 5–15% into bakery, meat analogs and dairy alternatives without sensory penalties, and chitin-derived chitosan films extend the bread shelf life by three days. Comparative life-cycle data indicate 45–80% lower greenhouse gas emissions and land use than equivalent animal-derived ingredients. Collectively, the evidence positions insect-based ingredients as versatile, safe, and climate-smart tools to enhance food quality and sustainability, while outlining research gaps in allergen mitigation, consumer acceptance, and regulatory harmonization. Full article

Incorporating health-promoting resveratrol into food products is challenging, primarily due to its poor solubility. Covalent conjugation is a promising, low-energy, and environmentally friendly strategy to overcome this limitation. This study compared the effectiveness of free radical grafting and alkaline methods for covalently conjugating whey protein isolate (WPI) with resveratrol. Conjugates were evaluated for molecular weight, structural characteristics, functional properties, and antioxidant activities. Both methods yielded conjugates with enhanced solubility relative to native resveratrol, with fold increases from 7.6 to 21.7 for the free radical grafting and from 8.1 to 23.6 for the alkaline method. Conjugates prepared via free radical grafting exhibited greater increases in molecular weight (10–100 kDa range), higher resveratrol incorporation (up to 17.6%), and superior functional properties compared to the alkaline conjugates (p < 0.05). Specifically, emulsifying activity, foaming capacity, and foaming stability improved by up to 64.7%, 45.8%, and 220.9%, respectively, compared to WPI. The antioxidant activities of the free radical grafting conjugates were 1.3- to 3.6-fold higher than those of alkaline conjugates. These findings highlight free radical grafting of WPI as a promising approach for incorporating resveratrol and improving the functionality of protein-based ingredients in functional food products. Full article

Innovative changes to our current food system are needed, and one solution is cultivated meat, which uses modern engineering, materials science, and biotechnology to produce animal protein. This article highlights the advantages of incorporating whey protein isolate (WPI) and β-lactoglobulin (β-LG) into hydrogel networks to aid cell growth on cultivated meat scaffolds. The protein and polysaccharide (i.e., alginate) components of the scaffolds are food-grade and generally regarded as safe ingredients, enabling the transition to more food-safe, edible, and nutritious scaffolds. The impact of WPI and varying properties on cell performance was evaluated; alginate concentration and the addition of proteins into the hydrogels significantly altered their stiffness and strength. The results of this study demonstrate the innocuous nature of novel scaffolds and reveal enhanced cell proliferation on WPI and β-LG-modified groups compared to standard biomaterial controls. This work serves as a stepping stone for more comprehensive analyses of WPI, β-LG, and alginate scaffolds for use in cultivated meat research and production. Full article

The development of bakeable foods supplemented with probiotics requires novel strategies to preserve the functionality of probiotic cells during thermal and gastrointestinal stress conditions. The objective of the present study was to evaluate the protective effect of multilayer double emulsions (W₁/O/W₂) stabilized with pectin-protein complexes on the viability of Limosilactobacillus reuteri (Lr) under thermal treatment (95 °C, 30 min), storage (4 °C, 28 d), and simulated gastrointestinal conditions. Emulsions were prepared with whey protein isolate (WPI) or sodium caseinate (Cas) as outer aqueous phase emulsifiers, followed by pectin coating and ionic gelation with calcium. All emulsions were stable and exhibited high encapsulation efficiency (>92%) with initial viable counts of 9 log CFU/mL. Double emulsions coated with ionically gelled pectin showed the highest protection against heat stress and gastrointestinal conditions due to the formation of a denser layer with lower permeability, regardless of the type of protein used as an emulsifier. At the end of storage, Lr viability exceeded 7 log CFU/mL in cross-linked pectin-coated microcapsules. These microcapsules maintained >6 log CFU/mL after thermal treatment, while viability remained >6.5 log CFU/mL during digestion and >5.0 log CFU/mL after consecutive heat treatment and simulated digestion. According to these results, the combination of double emulsion, multilayer formation and ionic crosslinking emerges as a promising microencapsulation technique. This approach offers enhanced protection for probiotics against extreme thermal and digestive conditions compared to previous studies that only use double emulsions. These findings support the potential application of this encapsulation method for the formulation of functional bakeable products. Full article

This study explored the development of a novel whey-based fermented beverage enriched with juice concentrates and health-promoting ingredients, emphasizing its bioactive properties. The formulation included whey protein isolate (5%), juice concentrates (10% apple, raspberry, and cranberry), and inulin (4%). Fermentation was carried out with the following strains: Lacticaseibacillus rhamnosus (LGG), Lacticaseibacillus casei (431), and Lactobacillus helveticus (R0052) at 2%. Antimicrobial activity was evaluated against pathogens including Listeria monocytogenes (strains 33423 and 33413), Staphylococcus aureus (113 and Newman), Bacillus cereus (DPC 6089), Escherichia coli (NCTC 9001), and Salmonella Enteritidis (NCTC 6676). Antioxidant capacity was measured using 2,2-Diphenyl-1-picrylhydrazylradical (DPPH) and Ferric Reducing Antioxidant Power (FRAP) assays, and angiotensin-converting enzyme (ACE) inhibitory activity was assessed. All bioactivities were found to be high in fermented whey beverage and a further significant increase was observed after simulated gastrointestinal digestion. This fruit-flavored whey beverage demonstrated notable antimicrobial and antioxidant activities, highlighting its potential for functional food applications aimed at combating harmful bacteria and oxidative stress. Full article

The effects of soy protein and whey protein supplementation on glycemic control show inconsistency, and the mechanisms underlying the impact of a high-protein diet on blood glucose regulation remain unclear. This study aimed to explore the impact of a dual-protein (DP) blend comprising soy protein isolate (SPI) and whey protein concentrate (WPC), processed through high-pressure homogenization, on mice with Type 2 diabetes mellitus (T2DM) and its potential mechanisms. In the in vitro experiments, an insulin-resistant (IR) HepG2 cell model was treated with DP, resulting in a significant enhancement of glucose uptake and upregulation of IRS1 and GLUT4 expression. For the in vivo experiments, male C57BL/6J mice were randomly assigned into four groups (n = 6) based on body weight: normal control, T2DM model group, Metformin-treated group, and DP-treated group. Following a 5-week feeding period, Metformin and DP significantly reduced levels of blood sugar, AUC, TC, TG, and LDL-C in T2DM mice. Additionally, TP and ALB levels in the DP group were notably higher in the model group. In the liver and pancreas, DP alleviated histopathological changes and promoted liver glycogen synthesis in T2DM mice. Moreover, the levels of IRS1 and PI3K in the livers of mice in the DP group were significantly higher than those in the model group. Compared with the model groups, DP significantly reduced the expression of CD45 and increased the expression of CD206 in the pancreas of mice. Furthermore, 16S rRNA analysis revealed that DP altered the composition of the gut microbiota in diabetic mice, increasing the relative abundance of Lactobacillus, Parvibacter, and Lactobacillaceae. This suggested that DP could alleviate functional metabolic disorders in the gut and potentially reverse the risk of related complications. In conclusion, soy whey dual-protein may have an effective nutritional therapeutic effect on T2DM mice by regulating lipid metabolism, the INS/IRS1/PI3K signaling pathway, and gut microbiota. Full article

Whey protein isolate (WPI) hydrogel is a promising candidate as a biomaterial for tissue engineering. Previously, WPI hydrogels containing poly-γ-glutamic acid (γ-PGA) with a molecular weight (MW) of 440 kDa demonstrated potential as scaffolds for bone tissue engineering. Here, the study compares different γ-PGA preparations of differing MW. WPI-γ-PGA hydrogels containing 40% WPI and 0%, 2.5%, 5%, 7.5%, and 10% γ-PGA were synthesised. Three γ-PGA MWs were compared, namely 10 kDa, 700 kDa, and 1100 kDa. Evidence of successful γ-PGA incorporation was demonstrated by scanning electron microscopy and Fourier transform infrared spectroscopy. Increasing γ-PGA concentration significantly improved the swelling potential of the hydrogels, as demonstrated by ratio mass increases of between 85 and 90% for each 10% variable group. Results suggested that γ-PGA delayed enzymatic proteolysis, potentially decreasing the rate of degradation. The addition of γ-PGA significantly decreased the Young’s modulus and compressive strength of hydrogels. Dental pulp mesenchymal stem cells proliferated on all hydrogels. The highest cellular growth was observed for the WPI-700 kDa γ-PGA group. Additionally, superior cell attachment was observed on all WPI hydrogels containing γ-PGA compared to the WPI control. These results further suggest the potential of WPI hydrogels containing γ-PGA as biomaterials for bone tissue engineering. Full article

Bone-associated pathologies are major contributors to chronic pathology statistics. Current gold standard treatments present limitations such as the ability to act as scaffolds whilst effectively delivering medications to promote cellular proliferation. Recent advancements in biomaterials have suggested whey protein isolate (WPI) hydrogel as a potential candidate to act as a scaffold with the capacity for drug delivery for bone regeneration. In this study, we investigate whey protein isolate hydrogels enhanced with the phytocannabinoid cannabidiol (CBD). The use of CBD in WPI hydrogels for bone regeneration is original. The results suggest that CBD was successfully incorporated into the hydrogels bound potentially through hydrophobic interactions formed between hydrophobic patches of the protein and the hydrophobic cannabinoid. The incorporation of CBD into the WPI hydrogels improved the mechanical strength of the hydrogels. The Young’s modulus was improved from 2700 kPa ± 117 kPa to 7100 kPa ± 97 kPa when compared to the WPI control, without plant-derived cannabinoids, to the WPI with the maximum CBD concentration. Furthermore, statistically significant differences for both Young’s modulus and compressive strength were observable between the WPI control and CBD hydrogel variables. The release of CBD from the WPI hydrogels was confirmed with the results suggesting a maximum release of 20 μM over the 5-day period. Furthermore, the hydrogels supported the proliferation and synthesis of collagen and calcium, as well as the alkaline phosphatase activity of MC3T3-E1 pre-osteoblasts, which demonstrates the potential of WPI/CBD hydrogels as a biomaterial for osseous tissue regeneration. 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

Biomaterials play a crucial role in the long-term success of bone implant treatment. The accumulation of bacterial biofilm on the implants induces inflammation, leading to implant failure. Modification of the implant surface with bioactive molecules is one of the strategies to improve biomaterial compatibility and limit inflammation. In this study, whey protein isolate (WPI) fibrillar coatings were used as a matrix to incorporate biologically active phenolic compound phloroglucinol (PG) at different concentrations (0.1% and 0.5%) on titanium alloy (Ti6Al4V) scaffolds. Successful Ti6Al4V coatings were validated by X-ray photoelectron spectroscopy (XPS), showing a decrease in %Ti and increases in %C, %N, and %O, which demonstrate the presence of the protein layer. The biological activity of PG-enriched WPI (WPI/PG) coatings was assessed using bone-forming cells, human bone marrow-derived mesenchymal stem cells (BM-MSCs). WPI/PG coatings modulated the behavior of BM-MSCs but did not have a negative impact on cell viability. A WPI with higher concentrations of PG increased gene expression relative to osteogenesis and reduced the pro-inflammatory response of BM-MSCs after biofilm stimulation. Autoclaving reduced WPI/PG bioactivity compared to filtration. By using WPI/PG coatings, this study addresses the challenge of improving osteogenic potential while limiting biofilm-induced inflammation at the Ti6Al4V surface. These coatings represent a promising strategy to enhance implant bioactivity. Full article