Structure-property relationship of pea protein fibrils in stabilization of HIPEs and the encapsulation, protection, controlled release and oral delivery of carotenoids for alleviating intestinal inflammation.

Increasing attentions are paid to high internal phase emulsions (HIPEs) due to their unique properties. In this study, pea protein-based fibrils were used as emulsifier to stabilize HIPEs. We demonstrated that the molecular assembly pathway and interfacial behavior of pea protein-based fibrils are affected by ionic strength. And the increased abundance of highly flexible worm-like nanofibrils facilitated their adsorption and packing on oil droplets, resulting in improved emulsion properties to stabilize the HIPEs with the internal phase volume fraction as high as 90%. Based on this, high loading content of carotenoids up to 0.05 wt% in the prepared HIPEs, protection of their stability against heating, UV and iron ions, and significantly increased bio-accessibilities of the carotenoids were realized. Animal studies using a mouse model of DSS-induced colitis revealed that carotenoid loaded HIPEs can alleviate the colon injury, by downregulating the expression of inflammatory cytokines, and promoting intestinal barrier function. This work will deepen the understanding of the formation of pea protein fibrils and provide a reference for the rational use of carotenoid loaded HIPEs in IBD management.

Protecting meat color: The interplay of betanin red and myoglobin through antioxidation and coloration.

Myoglobin (Mb) responsible for meat color is easily oxidized resulting in meat discoloration. Here, betanin red (BR), as a natural pigment and antioxidant, was chosen for enhancing redness and inhibiting oxidation. Multiple spectroscopies, isothermal titration calorimetry and molecular docking demonstrated that BR changed the microenvironment of heme group and amino acid residues of Mb, inhibited the oxidation of oxymyoglobin. The main interaction force was hydrogen bond and one variable binding site provided a continuous protective barrier to realize antioxidation. The combination of antioxidation with the inherent red color of BR offered dual color protection effect on processed beef with the addition amount of 0.2 % BR. BR treatment enhanced the redness by 25.59 âˆ¼ 53.24 % and the sensory acceptance by 4.89 âˆ¼ 14.24 %, and decreased the lipid oxidation by 0.58 âˆ¼ 15.92 %. This study paves a theoretical basis for the application of BR and its structural analogues in meat color protection and other quality improvement.

Hydrogels as promising carriers for the delivery of food bioactive ingredients.

The burden of public health challenges associated with the western dietary and living style is growing. Nutraceuticals have been paid increasing attentions due to their effects in promotion of health. However, in the gastrointestinal (GI) tract, the nutraceuticals suffer from not only the harsh acidic environment of the stomach and a variety of digestive enzymes, but also the antibacterial activity of intestinal bile salts and the action of protease from the gut microbiota. The amount of the nutraceuticals arriving at the sites in GI tract for absorption or exerting the bioactivities is always unfortunately limited, which puts forward high requirements for protection of nutraceuticals in a certain high contents during oral consumption. Hydrogels are three-dimensional polymeric porous networks formed by the cross-linking of polymer chains, which can hold huge amounts of water. Compared with other carries with the size in microscopic scale such as nanoparticle and microcapsules, hydrogels could be considered to be more suitable delivery systems in food due to their macroscopic bulk properties, adjustable viscoelasticity and large spatial structure for embedding nutraceuticals. Regarding to the applications in food, natural polymer-based hydrogels are commonly safe and popular due to their source with the appealing characteristics of affordability, biodegradability and biocompatibility. Although chemical crosslinking has been widely utilized in preparation of hydrogels, it prefers the physical crosslinking in the researches in food. The reasonable design for the structure of natural polymeric hydrogels is essential for seeking the favorable functionalities to apply in the delivery system, and it could be possible to obtain the enhanced adhesive property, acid stability, resistant to bile salt, and the controlled release behavior. The hydrogels prepared with proteins, polysaccharides or the mix of them to deliver the functional ingredients, mainly the phenolic components, vitamins, probiotics are discussed to obtain inspiration for the wide applications in delivery systems. Further efforts might be made in the in situ formation of hydrogels in GI tract through the interaction among food polymers and small-molecular ingredients, elevation of the loading contents of nutraceuticals in hydrogels, development of stomach adhesive hydrogels as well as targeting modification of gut microbiota by the hydrogels.

Purification of Egg White Lysozyme Determines the Downstream Fibrillation of Protein and Co-assembly with Phytochemicals to Form Edible Hydrogels Regulating the Lipid Metabolism.

Although the synthetic chemistry or synthetic biological systems have already shown the power of biomaterials engineering, natural bioresource matter is still a valuable library of raw ingredients for the production of biomaterials, in particular, the edible ones. However, the influence of upstream isolation and purification of the raw materials on their performance in the downstream processing procedures is still unexplored, which is essential for the engineering of biomaterials. Based on the comparison of conventional techniques, heating-induced precipitation combined with resin-blending ion exchange was developed as a simple and cheap method for the utilization of egg whites to produce the lysozyme that is found to be exclusively feasible for fibrillation. Even with similar purities, only the lysozyme prepared by this method could be utilized to form ordered linear aggregate fibrils. Fibrillation was recently pursued as a new approach to utilize bioresource mass for high-tech end-products. Phytochemicals, totally replacing salts, induced the lysozyme fibrils to form hydrogels spontaneously, which was further demonstrated in an in vivo study to prevent obesity induced by a high-fat diet (HFD) by reducing lipid absorption and lipogenesis, promoting energy expenditure, and inhibiting inflammation. The agri-food bioresource was successfully employed as a proof of concept in edible biomedical materials for the regulation of lipid metabolism.

Higher affinity of polyphenol to zein than to amyloid fibrils leading to nanoparticle-embed network wall scaffold to construct amyloid fibril-zein-EGCG hydrogels for coating of beef.

Globally, 800 million are undernourished and 2 billion are deficient in micronutrients. A regular dietary intake of meat is one of effective strategies to fight against undernourishment and deficiency of micronutrients. However, meat is a typical perishable food; proper handling to extend the shelf life of meat is required. In the present study, homogenous hydrogels composed with high contents of lysozyme amyloid fibrils, zein and green tea polyphenol epigallocatechin gallate (EGCG) were developed via the orchestration of protein-protein interaction and polyphenol-protein interaction for coating of beef. The protein-protein interaction between amyloid fibrils and zein showed higher affinity than that of the polyphenol-protein interactions. In addition, polyphenol EGCG was found to be more inclined to bind with zein than to the amyloid fibrils. Thus, the amyloid fibrils performed as the scaffold, in which EGCG on one hand induced the zein aggregate nanoparticles and on the other hand deposited on the surface of amyloid fibrils, leading to the dense multi-pore network with the nanoparticle-embed wall. It served as the microstructure mechanism for the enhanced gel strength. Coating of fresh chilling beef with the amyloid fibril-zein-EGCG hybrid hydrogels effectively protected the freshness and tenderness through inhibiting the over-growth of microorganisms and oxidation of lipid. This study paves the way to develop functional edible biomaterials via polyphenol induced coordination of the protein-protein interaction and polyphenol-protein interaction, where polyphenol acts as the molecular glue. This strategy shows high application potentials in health promotion related fields, including edible coating to extend the shelf-life of fresh meat.

The effects of high pressure treatment on the structural and digestive properties of myoglobin.

The effects of high pressure treatment (100-400 MPa for 20 min) on the structural and digestive properties of myoglobin were investigated by UV-vis absorption spectroscopy, intrinsic and synchronous fluorescence spectroscopy and molecular dynamics simulation. High pressure treatment induced the exposure of aromatic residues and changed the interaction between heme and globin, which in turn increased the gastrointestinal digestibility of myoglobin. Molecular dynamics simulation indicated that the hydrophobic interaction and hydrogen bonds of the myoglobin-pepsin complex were weakened after high pressure treatment, but the dipolar interaction was strengthened. The findings revealed the mechanisms on high pressure-induced increase in digestibility of myoglobin.

High Internal Phase Emulsions Stabilized with Polyphenol-Amyloid Fibril Supramolecules for Encapsulation and Protection of Lutein.

High internal phase emulsions (HIPEs), also called highly concentrated emulsions with a minimal internal phase volume fraction of 74%, have been paid increasing attention in the development of functional foods due to their high potential in loading with large amounts of hydrophobic nutriceuticals. In the present study, HIPEs stabilized by polyphenol-amyloid supramolecular filaments were prepared for encapsulation of olive oil and loading with lutein. Binding and stacking of the green tea polyphenol epigallocatechin gallate (EGCG) on the surface of amyloid fibrils fabricated from hen egg lysozyme resulted in the hybrid supramolecules, which assembled to form hydrogels. The amyloid fibril clusters shrouded by EGCG were observed in the microstructure of the hydrogels characterized by atomic force microscopy (AFM). HIPEs stabilized by the EGCG-amyloid fibril supramolecules showed the typical microstructure of highly packed polyhedral geometric oil droplets. The gel strength of the HIPEs stabilized by the hybrid supramolecules was greater than that of HIPEs stabilized by pure amyloid fibrils. The droplet size of the HIPEs first decreased and then increased with the increase of EGCG contents in the hybrid supramolecules, which was consistent with the corresponding emulsion morphologies obtained from the images of confocal laser scanning microscopy (CLSM). Aggregation of the protein-based nanofibrils appeared in the continuous phase at higher EGCG contents. The droplet size of the HIPEs decreased with the increase of the amyloid fibril concentration, accompanied by more packed and homogenously dispersed lipid droplets, as shown in the CLSM images. A high loading content of lutein of up to 10 mg/mL in the prepared HIPEs was realized, and the stability of lutein against ultraviolet irradiation, heat, iron, and hydrogen peroxide was promoted significantly. In addition, encapsulation with the HIPEs prevented the oxidization of olive oil, and this effect was enhanced with the increase of the EGCG content in the hybrid supramolecules ranging from 0 to 0.25 wt %. The protection function of the HIPEs might be ascribed to the membrane of interfacial amyloid fibrils and the crowded oil droplet environment, both of which could shield the pro-oxidation factors.

Ultrasound treatment can increase digestibility of myofibrillar protein of pork with modified atmosphere packaging.

We explored whether ultrasound treatment affected digestibility of myofibrillar protein (MP) isolated from modified atmosphere packed (MAP, 70% N2 and 30% CO2) pork. MP digestibility under pepsin and pancreatin treatments decreased significantly with storage time. Ultrasound treatment increased the digestibility and produced a greater number of smaller peptides. However, the total peptide count and unique peptide counts were significantly reduced. Moreover, active sulfhydryl, total sulfhydryl, particle size, turbidity and surface hydrophobicity of MP increased with time, while protein solubility and ATPase activity decreased. Compared with the untreated samples, ultrasound treatment increased protein solubility, surface hydrophobicity, and active sulfhydryl content, but decreased total sulfhydryl content (except 10 d), particle sizes, turbidity and ATPase activity. Ultrasound treatment resulted in a decline in ß-turn and α-helix contents. Therefore, ultrasound is conducive to the digestion. Additionally, structural and functional properties of protein in MAP were less stable than those in vacuum-packed pork reported before.

Interplay between transglutaminase treatment and changes in digestibility of dietary proteins.

The changes in digestibility of TG-treated myofibrillar protein (MP), soybean protein isolate (SPI) and mixed proteins were evaluated by measuring liberation of primary amino groups, monitoring structural changes and investigating peptide fingerprints. TG treatment generally increased gastric digestion of treated proteins, possibly due to the structural changes occurred during TG treatment. In contrast, the initial intestinal digestion was suppressed by TG treatment. Compared with MP, the digestibility and peptide composition of SPI were affected by TG treatment to a larger degree, possibly due to the higher level of glutamine in SPI. Peptidomics analysis indicated that the changes in peptide composition of digests of TG-treated samples were related with the loss of Lys residues during TG treatment. Larger quantities of bioactive peptides KIEFEQFLPM, EVHEPEEKPRPK and TVKEDQVFPMNPPK were released after digestion of TG-treated MP. These results highlighted the complex and substantial influence of TG treatment on the digestibility of dietary proteins.

Acetylation inhibition alleviates energy metabolism in muscles of minipigs varying with the type of muscle fibers.

In this study, we investigated whether preslaughter chemical-induced acetylation affected postmortem energy metabolism and pork quality. Thirty pigs were randomly assigned to control, acetyltransferase inhibitor (ATi) or deacetyltransferase inhibitor treatments. Serum, trapezius, longissimus lumborum, psoas major, semimembranosus and semitendinosus muscles were taken for analyses. The results indicated that ATi treatment significantly reduced the activities of lactate dehydrogenase and creatine kinase and heat shock protein 70 in serum (P < 0.05). ATi treatment increased ATP and glycogen content, but decreased lactic acid content in trapezius, psoas major and semitendinosus muscles (P < 0.05). A total of 13 acetylated proteins bands were identified and the deacetylation of creatine kinase may play a key role in slowing down the postmortem energy metabolism in ATi-treated group. In addition, ATi treatment reduced the rate of postmortem glycolysis in muscles with higher oxidative but lower glycolytic fibers. These findings provide a new insight into the underlying mechanism on muscle-specific postmortem changes of pork quality.

Effect of Sous-vide cooking on the quality and digestion characteristics of braised pork.

This study aimed to evaluate the effect of Sous-vide (SV) cooking on the quality, flavor and digestion characteristics of braised pork. Traditional (TD) sample had the highest fat content and malondialdehyde (MDA) value, but the lowest protein content and total sulfhydryl (SH) content (P < 0.05). The SH content in SV samples decreased with the heating time, but MDA content increased (P < 0.05). In addition, α-helix content of TD samples was higher but ß-sheet content was lower than SV samples. LC-MS/MS indicated that SV samples at 65 °C for 8 h had potential bioactive and antioxidant peptides. GC-MS mainly identified 395 volatile components and SV samples at 70 °C for 8 h and 75 °C for 8 h had similar flavor compounds to TD samples. Thus, Sous-vide cooking produced better quality and digestion characteristics of braised pork. These findings give a new insight into the associations of processing methods with meat quality.

Application of ultrasound treatment for improving the quality of infant meat puree.

Infant meat puree has an indispensable effect on the oral development and nutritional intake of infants. However, commercially available products have poor texture and relatively low digestibility. In this study, ultrasound (20 kHz and 200 W, 400 W, or 600 W) was applied to the pretreatment of raw meat for preparing infant meat puree for 15 min, 30 min, and 45 min. To assess the impact of ultrasound on infant meat puree, the viscosity, texture, water distribution, particle size and in vitro digestibility were determined. The results showed that, compared with control, viscosity and hardness of meat puree decreased and the texture was better in 400 W and 600 W groups. The content of immobilized water increased in comparison with the control. Ultrasound had no obvious effect on the digestibility of meat puree in gastric phase, but it increased the digestibility in intestinal phase with the highest digestibility (80.85%±3.33) in 600 W, 15 min group. Overall, the ultrasound parameters of 600 W for 15 min can be selected as the best condition to process infant meat puree. The findings provide a new perspective for the improvement of infant meat puree.

Changes of the Microbiota Composition on the Surface of Pig Carcasses during Chilling and Its Associations with Alterations in Chiller's Temperature and Air Humidity.

In this study, we investigated changes of microbiota composition on the surface of pig carcasses during chilling and their associations with temporal and spatial changes of wind speed, air temperature, and air humidity. The composition of microbiota on a carcass surface varied greatly with sampling sites; in particular, the surfaces of forelegs and neck had higher load of microorganisms and different microbiota composition compared to in the air and other carcass parts. However, such a difference in the microbiota composition decreased as chilling time extended. The positive detection ratios of microbial genes resistant to sulfonamides, quinolones, tetracyclines, and ß-lactams were found different greatly with chilling time and sampling sites. The ß-lactam and tetracycline resistant genes were observed in higher ratios in airborne microorganisms in the chiller, while the sulfa and tetracycline resistant genes had higher ratios in the microbiota on pig carcasses. Actual measurements and dynamic simulation showed that air temperature and humidity varied greatly among different places in a chiller within the first 8 h of chilling, with higher values close to the door, but the differences became smaller afterwards. The micro-environmental differences and changes in the chiller may cause the different composition of microbiota on pig carcasses.

Casein-fed mice showed faster recovery from DSS-induced colitis than chicken-protein-fed mice.

This study aimed to examine whether casein- and chicken protein-fed mice had different capacities of recovering from dextran sulfate sodium (DSS)-induced colitis. Mice were fed a chicken protein or casein diet for 14 days, which was followed by 7-day DSS treatment and then a 6-day recovery period by gavage of Akkermansia muciniphila (A. muciniphila). Compared with the chicken protein diet, the casein diet increased the relative abundance of beneficial gut bacteria, whereas DSS treatment did not induce significant differences in physiological and pathological indicators between the diet groups. During the recovery period, gavage of A. muciniphila alleviated colitis symptoms by decreasing the score of the disease activity index (DAI), spleen weight, and TNF-α mRNA level but increasing the mucus thickness and MUC2 mRNA level. Several genera, including the Ruminococcaceae NK4A214 group, Bifidobacterium, Roseburia, Ruminiclostridium and Lachnospiraceae NK4A136 group, may play a critical role. In addition, the casein diet helped DSS-treated mice recover faster from colitis, in terms of their body weight, colon length and histological score, probably due to its higher digestibility.

Influence of induction cooking on the flavor of fat cover of braised pork belly.

Fat has a great impact on texture and flavor of meat products, which is influenced by cooking methods. In this study, the profiles of fatty acids, volatile compounds, and texture of fat cover of braised pork belly were investigated after plane and concave induction cooking. The results showed that cooking time showed a great impact on fat content, textural properties, fatty acids composition, lipid oxidation, and volatile compounds of fat cover (p < 0.05). When cooking time was fixed, concave induction cooking caused lower hardness, chewiness, and saturated fatty acids but higher polyunsaturated fatty acids at 60 min than plane induction cooking. Electronic nose and GC-MS analyses showed that concave induction cooking had a greater impact on flavor of pork belly fat and produced a comparable flavor to plane induction cooked samples in a shorter time. Sensory evaluation showed that concave induction cooking had higher scores at 60 min. Thus, concave induction cooking could be a more efficient method for meat processing. PRACTICAL APPLICATION: Electromagnetic induction heating is an effective cooking technique. It is characterized by uniformity, efficiency, and safety of heating. The application of electromagnetic induction heating technology to the cooking of braised pork was studied, which provides information for further optimizing the cooking technology of braised pork and improving the quality of braised pork.

Ultrasonic treatment increased functional properties and in vitro digestion of actomyosin complex during meat storage.

We investigated the effects of ultrasonic treatment (400 W, 20 kHz, 45.52 W/L) and storage time (0 d, 3 d, 7 d and 10 d) on functional properties, structural changes and in vitro digestion of actomyosin complex isolated from vacuum-packed pork. As storage time increased, turbidity, surface hydrophobicity, active sulfhydryl and total sulfhydryl of actomyosin complex increased, while protein solubility decreased. Ultrasonic treatment increased surface hydrophobicity, protein solubility and active sulfhydryl content but decreased turbidity and total sulfhydryl content compared with the control. Ultrasonic treatment caused a reduction in α-helix content on 0 day and the fluorescence intensity of tryptophan and tyrosine residues. It increased pancreatin digestibility of actomyosin complex and the number of peptides of smaller than 1 kDa. However, it decreased the number of peptides. The findings provide a new insight into the application of appropriate ultrasonic treatment to promote meat digestibility.

Synergistic enhancement of loading contents and chemical stability of lycopene distributing both inside and on the oil/water interface.

Loading contents and chemical stability of lycopene were synergistically enhanced after dispersion in genipin-crosslinked-chitosan (CS) stabilized high internal phase emulsions (HIPEs). HIPEs could be prepared with the parameters for the emulsifiers of CS concentration from 0.5 to 5 mg/mL, pH value from 5.5 to 7.5, and CS/genipin mass ratio from 2:1 to 20:1. High loading content of lycopene, up to 0.25 wt% was achieved, with emulsifier in the final system only 1 mg/mL. As the loading contents were elevated, increasing amount of lycopene distributed in HIPEs in the form of insoluble crystals. Meanwhile, density of oil droplets decreased and the shape changed from polygon to sphere, which is supposed to be related to the interaction between the crystal and the oil-water interface. Stability of lycopene against ultraviolet, temperature, hydrogen peroxide, and iron ions was improved significantly, which could be ascribed to the layer of genipin-crosslinked-CS on oil droplet surface and the crystal status of lycopene. The storage stability of lycopene was improved tremendously after encapsulation by HIPEs. PRACTICAL APPLICATION: Low loading content of lycopene in emulsion systems is not conducive to the evaluation of its biological function in subsequent experiments, as well as their real application in food industry. It is also crucial to improve the stability of lycopene for the practical application in food industry. In this work, the loading content in delivery system and the chemical stability of lycopene are improved through encapsulation with high internal phase emulsions (HIPEs). The significance of these results may have implications in fields spanning from colloidal science to functional foods applications.

Overheating induced structural changes of type I collagen and impaired the protein digestibility.

Collagen, especially type I collagen, a major component for connective tissue in meat, determines the background tenderness and affects digestibility of meat. Heating may induce great changes in protein structure and its pepsin-treated digestion. The objective of this study was to investigate how heating affected type I collagen structure and in vitro pepsin-treated digestion. Type I collagen was heated at 60 °C, 70 °C, 80 °C for 0.5 to 2.5 h, and the spectrometric measurements and in vitro pepsin digestion were performed. Increased heating temperature caused the exposure of aromatic residues and an elevation of intensity of synchronous fluorescence spectra, but a reduction in the conformational stability of type I collagen (P < 0.05). Under the in vitro pepsin digestion, the Km value of enzymatic reaction increased as heating temperature rose, but overheating attenuated the affinity of type I collagen to pepsin. Heating at 70 °C for 0.5 h is good for type I collagen to get higher digestion.

Acetylation and Phosphorylation of Proteins Affect Energy Metabolism and Pork Quality.

Preslaughter handling has been shown to significantly affect meat quality, but the mechanisms are not fully understood. In this study, we investigated protein phosphorylation and acetylation in pig muscles at early postmortem time and their associations with meat quality attributes. Thirty pigs were randomly assigned to traditional (TH, n = 15) or mild handling (MH, n = 15). Compared with TH, MH reduced the incidence of pale, soft, and exudative (PSE) or dark, firm, and dry (DFD) pork. MH induced 65 and 20 peptides that match with 39 and 12 proteins to be more highly phosphorylated and acetylated, respectively. Creatine kinase, ß-enolase, α-1,4-glucan phosphorylase, tropomyosin, and myosin heavy chain isoforms 1, 4, and 7 were found to be simultaneously phosphorylated and acetylated, which may involve glycolysis, tight junctions, and muscle contraction. The phosphorylation and acetylation levels of differential proteins showed significant correlations with meat quality traits. These findings indicate that preslaughter MH can improve meat quality by regulating protein phosphorylation and acetylation involving energy metabolism in muscle.

Influence of salting process on the structure and in vitro digestibility of actomyosin.

Salting process is widely used in the process of meat products, whereas few studies have revealed the digestibility of actomyosin after salting treatment, which is closely related with the nutrition of meat. This work reported effect of salting on the structural change and digestibility of actomyosin before and after heat treatment. Actomyosin in 0.4 M and 0.8 M of NaCl had higher content of disulfide bonds, and actomyosin in 0.4 M NaCl showed the largest particle sizes before and after heat treatment. In addition, actomyosin in 0.6 M and 0.8 M of NaCl was oxidized more severely after heat treatment. Based on peptidomics analysis by using liquid chromatography tandem mass spectrometry (LC-MS/MS), actomyosin in 0.6 M was digested more easily, which was followed by sample in 0.8 M and 0.4 M of NaCl in descending order. The lowest digestibility of actomyosin in 0.4 M NaCl was related with its higher content of disulfide bond and severer aggregation behavior. The lower digestibility of actomyosin in 0.8 M NaCl should be related with the higher content of disulfide bonds and surface oxidation. These results highlight the crucial role of salting process in affecting the digestibility of meat protein.