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Personalized three-dimensional (3D) printed foods rich in probiotics were investigated. Lactiplantibacillus plantarum (Lp), as a representative of probiotics, was used to investigate the 3D printing of probiotic-rich dysphagia foods. Here, whey protein isolate nanofibrils (WPNFs) were coated and anchored on bacterial surfaces via biointerfacial supramolecular self-assembly, providing protection against environmental stress and the 3D printing process. The optimized composite gels consisting of High acyl gellan gum (0.25 g), whey protein isolate (1.25 g), fructooligosaccharides (0.75 g), Lp-WPNFs-Glyceryl tributyrate emulsion (Φ = 40%, 3.75 mL) can realize 3D printing, and exhibit high resolution, and stable shape. The viable cell count is higher than 8.0 log CFU/g. They are particularly suitable for people with dysphagia and are classified as level 5-minced & moist in the international dysphagia diet standardization initiative framework. The results provide new insights into the development of WPNFs-coating on bacterial surfaces to deliver probiotics and 3D printed food rich in probiotics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Whey protein nanofibrils (WPNFs) can be used in edible films and coatings (EFCs) because of its favorable functional properties, which rely on its well-ordered β -sheet structures, high hydrophobicity, homogeneous structure, and antioxidant activity. In the present study, WPNF-based edible coatings with glycerol (Gly) as plastic and titanium dioxide nanotubes (TNTs) as antimicrobial agents were studied. TNTs not only showed greater antibacterial activity than titanium dioxide nanoparticles (TNPs), but also increased interactions with WPNFs. The WPNF/TNT film had a smooth and continuous surface and was homogeneous with good mechanical properties. WPNF/TNT edible coatings (ECs) can help improve lipid peroxidation and antioxidant activity, limit microbial growth, reduce weight loss, and extend the shelf life of chilled beef. Given that the WPNF/TNT film components are low cost and show high antioxidant and antimicrobial activity, these optimized films have potential applications for various food products, including raw and chilled meat.

Here, we describe peptide aggregation, which is also known as enzymatic protein resynthesis. Whey protein hydrolysate (WPH) is the starting material for assembling peptides. Analyses of the involved amino acids, intrinsic fluorescence, fluorescence phase diagram, secondary structure, turbidity, and surface hydrophobicity were performed to investigate the reaction process. The aggregation mechanism consists of two parts: 1) formation and 2) aggregation of the building blocks that form the ordered secondary β-sheet structure. Constructing the building blocks requires at least one intermediate state, which is formed after 0.5 hours. Non-synergistic changes in the secondary and tertiary structures then allow the intermediate state to emerge.

The development of intelligent indicator film that can detect changes in food quality is a new trend in the food packaging field. The WPNFs-PU-ACN/Gly film was prepared based on whey protein isolate nanofibers (WPNFs). Anthocyanin (ACN) and glycerol (Gly) were used as the color indicator and the plasticizer, respectively, while pullulan (PU) was added to enhance mechanical properties of WPNFs-PU-ACN/Gly edible film. In the study, the addition of ACN improved the hydrophobicity and oxidation resistance of the indicator film; with an increase in pH, the color of the indicator film shifted from dark pink to grey, and its surface was uniform and smooth. Therefore, the WPNFs-PU-ACN/Gly edible film would be suitable for sensing the pH of salmon, which changes with deterioration, as the color change of ACN was completely consistent with fish pH. Furthermore, the color change after being exposed to grey was evaluated in conjunction with hardness, chewiness, and resilience of salmon as an indication. This shows that intelligent indicator film made of WPNFs, PU, ACN, and Gly could contribute to the development of safe food. [ABSTRACT FROM AUTHOR]

Pro-Glu-Trp (PEW) and Leu-Leu-Trp (LLW) are peptides derived from whey protein digestive products; both peptides exhibit xanthine oxidase inhibitory activity in vitro. However, it remains unclear whether these peptides can alleviate hyperuricemia (HUA) in vivo. In this study, we investigated the roles of PEW and LLW, both individually and in combination, in alleviating HUA induced by potassium oxonate and hypoxanthine. Together, PEW and LLW exhibited synergistic effects in reducing the serum levels of uric acid (UA), creatinine, and blood urea nitrogen, as well as increasing the fractional excretion of UA. The combined treatment with PEW and LLW inhibited UA synthesis, promoted UA excretion, and restored renal oxidative stress and mitochondrial damage. Moreover, the combined treatment alleviated dysbiosis of the gut microbiota, characterized by increased helpful microbial abundance, decreased harmful bacterial abundance, and increased production of short-chain fatty acids. Taken together, these results indicate that the combination of PEW and LLW mitigate HUA and kidney injury by rebalancing UA synthesis and excretion, modulating gut microbiota composition, and improving oxidative stress. [Display omitted] [ABSTRACT FROM AUTHOR]

Pro-Glu-Trp (PEW), a whey protein-derived peptide, has been previously shown in vitro to have anti-hyperuricemic potential. The current study further evaluated the roles and the underlying mechanism of PEW in the management of hyperuricemia (HUA) in rat induced by potassium oxonate (PO) and hypoxanthine. Results revealed that PEW significantly reduced the levels of uric acid (UA), creatinine (Cr), and blood urea nitrogen (BUN) in serum, and effectively suppressed the activities of xanthine oxidase (XOD) associated with UA synthesis and modulated the expression of organic ion transporters related to UA excretion. Moreover, PEW alleviated UA-induced renal inflammation by regulating oxidative stress, suppressing the level of pro-inflammatory cytokines, and inhibiting the activation of NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome and Toll-like receptor 4/myeloid differentiation factor 88/NF-kappaB (TLR4/MyD88/NF-κB) signaling pathway. Taken together, these relults indicated that PEW improved HUA and renal inflammation by inhibiting UA synthesis, promoting renal UA excretion and suppressing NLRP3 inflammasome and TLR4/MyD88/NF-κB signaling pathways. [ABSTRACT FROM AUTHOR]

Whey protein isolate nanofibrils (WPNFs) can be used as a novel stabilizer in the Pickering emulsion system to improve the water solubility, stability and bioavailability of lipophilic bioactive ingredients. In this study, conjugated linoleic acid (CLA) and WPNFs were used to prepare a stable Pickering emulsion. We used a transmission electron microscope, low-temperature scanning electron micrographs and other methods to evaluate the micromorphology, surface hydrophobicity and structural units of the obtained WPNFs. Compared with whey protein isolate/CLA Pickering emulsion, the WPNFs/CLA Pickering emulsion has greater ability to remove 2,2-Diphenyl-1-picrylhydrazyl and 2,2′-amino-di(2-ethyl-benzothiazoline sulphonic acid-6) ammonium salt free radicals. Furthermore, the WPNFs/CLA Pickering emulsion has a more stable effect in terms of droplet size and zeta potential over a wider range of ionic strength and temperature conditions. These findings indicate that Pickering emulsion stabilized by WPNFs is more suitable as a carrier of CLA, as it increases the solubility of CLA and has better active applications in biology and food. [ABSTRACT FROM AUTHOR]

Salted duck egg yolk (SDEY) is one of the traditional pickled egg products in Asian countries, which suffers from the weight loss and deterioration of texture characteristics during storage. To better maintain the texture of SDEY, an edible coating based on whey protein isolate nanofibers (WPNFs) with glycerol (Gly) as a plasticizer and incorporating carvacrol (CA) as an antimicrobial agent was developed. Whey protein isolate (WPI, 5%) was used to self-assemble into WPNFs at 80 °C for 10 h. The particle size, zeta-potential and microstructure of WPNFs–CA emulsion were investigated to evaluate the distribution. Results proved that WPNFs–CA emulsion had smaller particle size and better distribution than WPI–CA emulsion. WPNFs–CA/Gly edible coating was then prepared based on WPNFs–CA emulsion. The WPNFs–CA/Gly edible coating exhibited higher antibacterial activity while the WPNFs–CA/Gly film had smooth and continuous surfaces and better transmittance compared with other samples. Furthermore, weight losses and textural properties changes of SDEYs with WPNFs–CA/Gly coating were evaluated. Results proved that salted duck egg yolks with WPNFs–CA/Gly coating exhibited lower weight losses. Textural properties were significantly improved by the WPNFs–CA/Gly coating on SDEYs than those uncoated samples. It was noted that the egg yolks coated with the WPNFs–CA/Gly coating had the lowest hardness increase rate (18.22%). Hence, WPNF-based coatings may have a good development prospect in the food industry. [ABSTRACT FROM AUTHOR]

The composition and properties of edible coatings (ECs) will significantly influence their effects of food preservation. For the first time, whey protein isolates nanofibers (WPNFs), as a novel material with high hydrophobicity and antioxidant activity, combined with carvacrol (CA) as an antimicrobial agent and glycerol (Gly) as a plasticizer, was used to prepare edible coating (WPNFs-CA/Gly) for preserving fresh-cut Cheddar cheese. The prepared WPNFs and ECs emulsions have been investigated with transmission electron microscopy. Furthermore, the antioxidant activity of ECs emulsions, antimicrobial activity of edible films, and the physical properties of edible films, such as micromorphology, thickness, transparency, and moisture content, have also been evaluated. The weight losses and physical characteristics of both coated and uncoated fresh-cut Cheddar cheese samples have been assessed during storage. The DPPH free radical scavenging rate of WPNFs-CA/Gly emulsion was up to 67.89% and the reducing power was 0.821, which was higher than that of WPI-CA/Gly emulsions. The antimicrobial activity of WPNFs-CA/Gly films was nearly 2.0-fold higher than that of WPNFs/Gly films for the presence of CA. The WPNFs-CA/Gly films had smooth and continuous surfaces, and the transparency reached 49.7% and the moisture content was 26.0%, which was better than that of WPI-CA/Gly films. Furthermore, Cheddar cheese with WPNFs-CA/Gly coatings has shown lower weight losses (15.23%) and better textural properties than those uncoated samples. This in-depth study has provided a valuable and noteworthy approach about the novel edible coating material. [ABSTRACT FROM AUTHOR]

At present, peptides are separated by molecular exclusion chromatography and liquid chromatography. A separation method is needed in any case, which can be scaled up for industrial scale. In this study, aqueous two-phase extraction (ATPE) and aqueous two-phase flotation (ATPF) were applied to separate and enrich antioxidant peptides from trypsin hydrolysates of whey protein isolates (WPI). The best experimental conditions were investigated, and the results were evaluated using the 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) free radical scavenging activity of the peptides-per-unit concentration and the recovery rate (Y) of peptides in the top phase of both ATPE and ATPF. Under optimal conditions, the Y and ABTS free radical scavenging activity per unit concentration in top phase of ATPE could reach 38.75% and 12.94%, respectively, and in ATPF could reach 11.71% and 29.18%, respectively. The purified peptides were characterized by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and reversed-phase high-performance liquid chromatography (RP-HPLC). PeptideCutter and PeptideMass were applied to analyze and calculate the peptide sequencing. KILDKVGINYWLAHK, VGINYWLAHKALCSEK, and TPEVDDEALEKFDKALK sequences having antioxidant activity were detected in the top phase of ATPE, and VGINYWLAHKALCSEK, KILLDKVGINYWLAHK, ILLDKVGINYWLAHK, IIAEKTKIPAVFK, KIIAEKTKIPAVFK, and VYVEELKPTPEGDLEILLQK sequences having antioxidant activity were detected in the top phase of ATPF. In conclusion, antioxidant peptides were successfully separated from the WPI hydrolysate by ATPE and ATPF; compared with ATPE, ATPF has superior specificity in separating antioxidant peptides. [ABSTRACT FROM AUTHOR]

Whey protein isolates (WPI) have been studied for film formation because of their favorable functional properties and industrial surplus; however, little is known regarding whey protein nanofibrils (WPNFs) as raw material for edible films. In the present study, WPNF-based edible coatings plasticized with glycerol (Gly) and trehalose (Tre) were studied. WPI (5%) can self-assemble into nanofibrils at 80 °C for 10 h under 220 rpm constant magnetic stirring. Compared with that of WPI, the hydrophobic and antioxidant activities of WPNFs were significantly increased. The microstructures and functions of WPI/Gly, WPNF/Gly, and WPNF/Gly/Tre films were characterized by scanning electron microscopy and atomic force microscopy, and by analyzing their contact angles, film thickness, transparency, moisture content, and solubility in water. Results showed that the formation of WPNFs increased surface smoothness, homogeneity, continuity, hydrophobicity, and transparency, and decreased the moisture content and water solubility of the films. To evaluate the performance of the edible coating, coated fresh cut apples were analyzed during storage. We found that the coatings containing WPNF (5%, w/v), Gly (4%, w/v) and Tre (3%, w/v) afforded the best protective action toward retarding the total phenolic content, browning, and product weight loss. Overall, the low cost, high biocompatibility, and consumer acceptance in taste tests support the ready application of these optimized WPNF films for commercial purposes. [ABSTRACT FROM AUTHOR]

Nanofibrils have favorable functional properties that facilitate their applications in the food industry. Whey protein isolate (WPI) is widely used to prepare nanofibrils via acid hydrolysis. In this research, the effect of hydrolysis by endoproteinase GluC (GluC) on WPI nanofibrils (WPNFs) was evaluated based on their structural and emulsifying properties. Following the hydrolysis of WPI by GluC at 37 °C and pH 8.0, the peptides self-assembled into WPNFs at 37 °C and pH 2.0. WPNFs with several micrometric length and nanometric (1–10 nm) diameter exhibited high aspect ratio (length/diameter). Based on microscopic images, circular dichroism spectroscopy, and infrared spectroscopy, we concluded that β-sheets played an important role in the formation and stability of WPNFs. The main factor affecting WPNF formation was found to be hydrolysis by GluC for 10 h followed by aggregation at pH 2.0. Whey protein hydrolysates with glutamic acid or aspartic acid residues, obtained from WPI hydrolysis by GluC for 10 h, might be positively charged in an acidic environment, thereby favoring the formation of ordered aggregates instead of random ones. WPNFs prepared under such conditions have a strong ability of improving the oxidative stability, activity and stability of emulsions. This new strategy for producing WPNFs will potentially expand their application in the food industry. Image 1075969 • WPNFs were prepared by hydrolyzing WPI with endoproteinase GluC. • The effect of endoproteinase GluC hydrolysis on WPNF formation was studied. • Building blocks of WPNFs were analyzed. • WPNF emulsions showed oxidative stability. • WPNFs showed emulsifying/stabilizing properties. [ABSTRACT FROM AUTHOR]

Personalized ready-to-eat foods with complex three-dimensional (3D) structure can be realized through 3D printing. As a representative of high-fat food, Cheddar cheese has been selected to investigate the effect of Whey protein isolate nanofibrils (WPNFs) to reduce fat content and improve 3D printability. With excellent emulsifying, thickening, gelling and antioxidant activities, WPNFs have been used as the only additive to prepare processed cheese. As observed in the CLSM micrographs and LF-NMR analysis, WPNFs can immobilize the increased moisture and homogenize the distribution of fat and moisture. The cohesiveness of WPNF-processed cheese has been proved to be a key factor in maintaining line continuity and layer-to-layer fusion during 3D printing. In addition to regulating the printability of processed cheese, WPNFs could delay the color change of 3D printed constructions for its antioxidant properties. The 3D rabbit and swan printed by F20 have showed clear details and accurate shape, smooth surfaces, and stability. This study will provide a novel food additive WPNFs to improve the 3D printability of high-fat foods with customized shape, texture, flavor, and nutritional requirements. [Display omitted] • WPNFs can efficiently improve 3D printability of high-fat food. • WPNFs better homogenized the distribution of fat and water in processed cheeses. • WPNF-processed cheeses showed improved 3D printing accuracy and stability. [ABSTRACT FROM AUTHOR]

The present work aimed to explore nanofibrils prepared from whey protein isolate (WPNFs) by hydrothermal to stabilize high internal phase Pickering emulsion (HIPPEs). WPNFs with well-ordered β -sheet structures and high aspect ratio were efficiently prepared by hydrothermal at 110 °C for 4 h, which is more efficiently than conventional method at 80–90 °C for 5–24 h. The characteristics of the building blocks analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with Tris-Tricine buffer system and liquid chromatography linked to tandem mass spectrometry (LC-MS/MS) proved that the formation of WPNFs is based on the hydrolysis of WPI, and hydrothermal can make WPI hydrolysis faster and more efficiently. The high internal phase Pickering emulsion (HIPPEs) stabilized by 5 wt% of WPNFs (internal phase fraction Φ up to 0.90) showed an excellent stability. The microstructure of HIPPEs illustrated by super resolution microscope and cryo - scanning electron microscopy showed that WPNFs tightly wrapped the oil droplets forming an ordered viscoelastic and dense interface layer. It can be speculated that WPNFs were flexible due to their large aspect ratio and mechanical strength, which allowed them to bend on the surface of the droplet to form a better package protection. Rheology and texture analysis showed that WPNFs stabilized HIPPEs have solid-like and tunable rheological properties. WPNFs were verified to improve the oil binding capacity and antioxidant activity. The results provided that hydrothermal is a novel and efficient method for preparing WPNFs, and the prepared WPNFs can be used to stabilize HIPPEs with tunable rheology. [Display omitted] • WPNFs efficiently prepared by hydrothermal at 110 °C for 4 h. • As low as 5% WPNFs could stabilize HIPPEs (Φ = 0.9). • The HIPPEs showed improved oil oxidation stability. • The HIPPEs with a tunable rheology was an alternative to margarine and solid fats. [ABSTRACT FROM AUTHOR]