A novel EGCG-Histidine complex improves gelling and physicochemical properties of porcine myofibrillar proteins: Insight into underlying mechanisms.

Herein, an EGCG-Histidine complex is prepared, characterized, and further used to improve gel properties of myofibrillar proteins (MP). Results of FTIR, XRD, UV-Vis spectroscopy showed that histidine is covalently bound to EGCG by Michael addition or Schiff base reaction to form EGCG-Histidine complex, and antioxidant activity of EGCG-Histidine complex is significantly increased compared to EGCG or histidine alone (P < 0.05). The addition of EGCG-Histidine complex results in cooking loss of gel decreasing from 66.7 ± 0.23 % to 40.3 ± 2.02 %, and improves rheological properties of MP, and enhances gel strength from 0.10 ± 0.01 N to 0.22 ± 0.03 N, indicating positive effect of EGCG-Histidine complex on MP gel formation, above results is supported by results of SEM, CD spectroscopy, SDS-PAGE, and tryptophan fluorescence. These results indicated that EGCG-Histidine complex can be used as a functional ingredient to improve gel quality of meat products.

Effect of portulaca (Portulaca oleracea L.) extract on the quality and physicochemical attributes of vacuum-packed seasoned steaks during chilled storage.

BACKGROUND: Vacuum packaging has the ability to reduce oxidative deterioration and microbial-induced spoilage of meat. However, in an oxygen-free environment, it can lead to the development of an unappealing purplish-red color and a decrease in the water-holding capacity of meat, thereby impacting the overall meat quality. Portulaca oleracea L. (POL) is a homology of medicine and food known for its exceptional antioxidant and antimicrobial properties. RESULTS: The aim of our present study was to investigate the antioxidant and antimicrobial ability of n-butanol phase extract of POL and the effect of POL extract incorporation on the quality (water-holding capacity, shear force, color, and texture) and physicochemical (pH, total volatile base nitrogen, and total viable counts) attributes of vacuum-packed seasoned steaks at 4 °C over a 15-day period. Results showed that the POL extract had excellent antioxidant and antimicrobial capacity. Furthermore, the addition of POL extract significantly inhibited protein oxidation and microbial growth of steaks (P < 0.05), and improved the water-holding capacity, color properties, and tenderness (P < 0.05). Moreover, there were no significant differences (P > 0.05) in the color, water-holding capacity, or tenderness between the 0.5 and 1 g kg-1 POL extract treatment groups compared to the sodium nitrite control group. CONCLUSION: These results indicate that POL extract had the potential to replace sodium nitrite due to its ability to hinder protein oxidation and microbial growth of vacuum-packed seasoned steaks, while enhancing the color, water-holding capacity, and tenderness of seasoned steaks. © 2024 Society of Chemical Industry.

Superabsorbent whey protein isolates/chitosan-based antibacterial aerogels: Preparation, characterization and application in chicken meat preservation.

Traditional absorbent pads are composed of hard-to-degrade polyethylene film and non-woven bottom layer, which have the characteristic of low absorption rate, without antibacterial effect. The objective of this study is to fabricate a novel superabsorbent and antibacterial aerogel, which consists of whey protein isolate (WPI) and chitosan (CS). The citric acid (CA) and ε-polylysine hydrochloride (ε-PLH) are incorporated into WPI/CS-based aerogel as cross-linking and antibacterial agent, respectively. The application in meat preservation as an absorbent pad is investigated. Results of water absorption, water vapor absorption and stress-strain show that aerogel comprised of 6 % WPI, 1.2 % CS, 2.0 % CA, and 2.0 % ε-PLH have the best water absorption capacity and stress. The density of WPI/CS/CA/ε-PLH aerogel is 82.7 ± 6.4 mg/cm3, and has a uniform and polyporous microstructure, resulting in superabsorbent capacity. Antibacterial rate of WPI/CS/CA/ε-PLH aerogel against Staphylococcus aureus, Escherichia coli, Salmonella and Listeria monocytogenes reach around 80 %. The WPI/CS/CA/ε-PLH aerogel significantly reduces increased velocity of b⁎, pH, total volatile base nitrogen, and total viable counts and decreased velocity of L⁎ and b⁎ of chicken meat (P < 0.05). Results indicate WPI/CS/CA/ε-PLH aerogel effectively extends shelf-life of chicken meat to 7 days, and could be used as an absorbent pad in meat preservation.

Generation of three-dimensional meat-like tissue from stable pig epiblast stem cells.

Cultured meat production has emerged as a breakthrough technology for the global food industry with the potential to reduce challenges associated with environmental sustainability, global public health, animal welfare, and competition for food between humans and animals. The muscle stem cell lines currently used for cultured meat cannot be passaged in vitro for extended periods of time. Here, we develop a directional differentiation system of porcine pre-gastrulation epiblast stem cells (pgEpiSCs) with stable cellular features and achieve serum-free myogenic differentiation of the pgEpiSCs. We show that the pgEpiSCs-derived skeletal muscle progenitor cells and skeletal muscle fibers have typical muscle cell characteristics and display skeletal muscle transcriptional features during myogenic differentiation. Importantly, we establish a three-dimensional differentiation system for shaping cultured tissue by screening plant-based edible scaffolds of non-animal origin, followed by the generation of pgEpiSCs-derived cultured meat. These advances provide a technical approach for the development of cultured meat.

Tea polyphenols coated sodium alginate-gelatin 3D edible scaffold for cultured meat.

This study aimed to use edible scaffolds as a platform for animal stem cell expansion, thus constructing block-shaped cell culture meat. The tea polyphenols (TP)-coated 3D scaffolds were constructed of sodium alginate (SA) and gelatin (Gel) with good biocompatibility and mechanical support. Initially, the physicochemical properties and mechanical properties of SA-Gel-TP scaffolds were measured, and the biocompatibility of the scaffolds was evaluated by C2C12 cells. SEM results showed that the scaffold had a porous laminar structure with TP particles attached to the surface, while FT-IR results also demonstrated the encapsulation of TP coating on the scaffold. In addition, the porosity of all scaffolds was higher than 40% and the degradation rate during the incubation cycle was less than 40% and the S2-G1-TP0.1-3 h scaffold has excellent cell adhesion and extension. Subsequently, we inoculated rabbit skeletal muscle myoblasts (RbSkMC) on the scaffold and induced differentiation. The results showed good adhesion and extension behavior of RbSkMC on S2-G1-TP0.1-3 h scaffolds with high expression of myogenic differentiation proteins and genes, and SEM results confirmed the formation of myotubes. Additionally, the adhesion rate of cells on scaffolds with TP coating was 1.5 times higher than that on scaffolds without coating, which significantly improved the cell proliferation rate and the morphology of cells with extension on the scaffolds. Furthermore, rabbit-derived cultured meat had similar appearance and textural characteristics to fresh meat. These conclusions indicate the high potential of the scaffolds with TP coating as a platform for the production of cultured meat products.

Changes in the Quality of Myofibrillar Protein Gel Damaged by High Doses of Epigallocatechin-3-Gallate as Affected by the Addition of Amylopectin.

This work investigated the improvement of amylopectin addition on the quality of myofibrillar proteins (MP) gel damaged by high doses of epigallocatechin-3-gallate (EGCG, 80 µM/g protein). The results found that the addition of amylopectin partially alleviated the unfolding of MP induced by oxidation and EGCG, and enhanced the structural stability of MP. Amylopectin blocked the loss of the free amine group and thiol group, and increased the solubility of MP from 7.0% to 9.5%. The carbonyl analysis demonstrated that amylopectin addition did not weaken the antioxidative capacity of EGCG. It was worth noting that amylopectin significantly improved the gel properties of MP treated with a high dose of EGCG. The cooking loss was reduced from 51.2% to 35.5%, and the gel strength was reduced from 0.41 N to 0.29 N after adding high concentrations of amylopectin (A:E(8:1)). This was due to that amylopectin filled the network of MP gel after absorbing water and changed into a swelling state, and partially reduced interactions between EGCG and oxidized MP. This study indicated that amylopectin could be used to increase the polyphenol loads to provide a more lasting antioxidant effect for meat products and improve the deterioration of gel quality caused by oxidation and high doses of EGCG.

Gellan gum-gelatin scaffolds with Ca2+ crosslinking for constructing a structured cell cultured meat model.

As an emerging technology to obtain protein by culturing animal-derived cells in vitro, it is crucial to construct 3D edible scaffolds to prepare structured cell cultured meat products. In this study, a scaffold based on gellan gum (GG)-gelatin (Gel) was prepared and further cross-linked with Ca2+. FTIR confirmed the electrostatic interaction between GG and Gel and the ionic cross-linking of Ca2+ and carboxyl groups, and SEM images showed the porous structure of the scaffolds. The staining results showed that scaffolds with high concentrations of Ca2+ had higher biocompatibility than scaffolds with low concentrations of Ca2+ and non-crosslinked scaffolds, and scaffolds Ca2+-GG2-Gel3-0.5 adhered to more cells and were more conducive to cell spreading. The immunofluorescence staining, SEM images, Western blot, and RT-qPCR showed that the scaffolds supported the proliferation and myogenic differentiation of chicken skeletal muscle satellite cells (CSMSCs) and myotubes were formed on the scaffolds. Finally, the scaffolds were stained and fried after culturing. The results of the textural and chromatic analysis showed that the texture and color of the scaffolds were similar to fresh meat and meat products. These results showed that ionically crosslinked GG-Gel scaffolds are biocompatible and stable for structured cell cultured meat models.

Suppression mechanism of L-lysine on the Epigallocatechin-3-gallate-induced loss of myofibrillar protein gelling potential.

As a natural antioxidant, Epigallocatechin-3-gallate (EGCG) needed to be added in high doses to maintain the quality of meat products. However, high doses of EGCG caused the excessive aggregation of myofibrillar protein (MP), which damaged the gel properties of MP gels. Therefore, the purpose of this study was to investigate the mitigation of EGCG-induced loss of MP gelling potential by L-Lysine (L-Lys). The results showed that the addition of 20 mM L-Lys induced excessive unfolding and loose aggregation of MP at 10 µmol/g EGCG, and hence, reducing the solubility (14.5%) and the tryptophan fluorescence, and forming a network structure with a large aperture. Therefore, the cooking loss was decreased from 29.20% to 15.13%, and the strength of MP gels was decreased from 0.35 N to 0.17 N. However, L-Lys hindered the hydrogen bonding interactions and hydrophobic interactions between MP and EGCG by competing the binding sites of MP at 50 µmol/g EGCG, which was supported by the Zeta potential, surface hydrophobicity, FTIR and molecular docking analysis. Thus L-Lys mitigated the protein aggregation caused by 50 µmol/g EGCG, improved the solubility (23.02%∼86.99%) and apparent viscosity, which were beneficial for the formation of a continuous network structure in MP gels. Therefore, the cooking loss of MP gels was decreased from 52.40% to 41.30%, and the gel strength was enhanced from 0.13 N to o.22 N with 20 mM L-Lys addition. The present study could provide a new strategy for increasing the amounts of EGCG in meat products without damaging the gel properties of meat products.

Incorporating Portulaca oleracea extract endows the chitosan-starch film with antioxidant capacity for chilled meat preservation.

This study aimed to investigate the application potential of Portulaca oleracea extract (POE) in active packaging for the preservation of chilled meat. First, the antioxidant capacity and active ingredients of POE were systematically studied. The results demonstrated that POE has excellent antioxidant capacity and contains abundant antioxidant compounds. Subsequently, antioxidant-active packaging films based on chitosan and starch containing different concentrations of POE (CS/POE films) were successfully developed. The main physicochemical and mechanical properties of the CS/POE films were characterized and evaluated. The CS/POE films exhibited remarkable antioxidant activity and can significantly reduce lipid oxidation in meat. Compared with polyethylene film, the CS/POE films-treated meats had better preservation effects and longer shelf-life. These findings suggested that CS/POE film has the potential to become a good alternative to conventional plastics in food packaging. In conclusion, Portulaca oleracea extract is an excellent natural antioxidant with great potential in active packaging for chilled meat preservation.

Effects of heating rates on the self-assembly behavior and gelling properties of beef myosin.

BACKGROUND: Myosin is the most important component of myofibrillar protein, with excellent gelling properties. To date, heating treatment remains the mainstream method for forming gel in meat products, and it has the most extensive application in the field of meat industry. However, at present, there are few reports on the effects of heating rates on myosin self-assembly and aggregation behavior during heating treatment. RESULTS: The present study aimed to investigate the effects of different heating rates (1, 2, 3 and 5 °C min-1 ) on the self-assembly behavior, physicochemical, structural and gelling properties of myosin. At the lowest heating rate of 1 °C min-1 , the myosin gel had a dense microstructure, the highest elastic modulus (G') and water holding capacity compared to higher heating rates (2, 3 and 5 °C min-1 ). At higher temperatures (40, 45 °C), the surface hydrophobicity, turbidity, particle size distribution and self-assembly behavior of myosin in pre-gelling solutions showed that myosin had sufficient time to denature, underwent full structure unfolding before aggregation at the heating rate of 1°C min-1 , and formed regular and homogeneous spherical aggregates. Therefore, the myosin gel also had a better three-dimensional network. CONCLUSION: The heating rates had an important effect on the quality of myosin gels, and had theoretical implications for improving the quality of meat gel products. © 2023 Society of Chemical Industry.

Improved thermal tolerance of ovotransferrin against pasteurization by phosphorylation.

Ovotransferrin is the most heat-sensitive protein in egg white which alters the processing suitability. To improve thermostability, phosphorylation modification was performed under different pH values (5.0, 6.0, 7.0, 8.0, and 9.0), and the reasons for which were explored via physicochemical changes in this study. SEM and particle size results showed that phosphorylation diminished the average particle diameter and the number of attached particles. CD results revealed that the α-helix content of phosphorylated ovotransferrin increased and the random coil decreased significantly (P < 0.05). Tertiary structure, surface hydrophobicity and ζ-potential analyses showed that phosphorylation made more hydrophobic amino acids buried inside ovotransferrin. These changes might be because most of the phosphorylation occurred in α-helix and ß- sheet to promote the thermal tolerance of phosphorylated ovotransferrin by improving its structure orderliness and decreasing surface hydrophobicity. The results indicated that phosphorylation was a practical method to enhance the thermal stability of heat-sensitive ovotransferrin.

Effects of NaCl on the interactions between neomethyl hesperidin dihydrochalcone and pork myofibrillar protein: Their relevance to gelation properties.

Effects of sweetener neomethyl hesperidin dihydrochalcone (NHDC) on the gelation properties of myofibrillar protein (MP) at 0.2 M and 0.6 M NaCl were investigated. The results showed that addition of NHDC did not obviously change the cooking loss and strength of MP gels at 0.2 M NaCl. Whilst at 0.6 M NaCl, addition of high doses of NHDC (50-100 µM/g) remarkably increased cooking loss and decreased strength of MP gels. The expanded MP enhanced the interactions between NHDC and MP, leading to aggregation of MP as evidenced by the increasing particle size and the reducing solubility. The MP aggregates impeded hydrophobic interactions and disulfide bonds crosslinking in the gelation process, thereby damaged the gelation properties of MP at 0.6 M NaCl. This was supported by the poor network of MP gels observed by SEM. In the end, molecular docking elucidated the main types of interaction at molecular level, including hydrogen bonding and hydrophobicity. The results would provide guidance for NHDC as a potential sweetener in meat products.

An injectable antibacterial chitosan-based cryogel with high absorbency and rapid shape recovery for noncompressible hemorrhage and wound healing.

Great challenges remain in the effective control of irregular and incompressible deep wound bleeding and the promotion of wound healing after bacterial infection. In this study, cryogels were prepared using an ice template based on chitosan (CS), oxidized gallic acid (OGA) and hemin (HE), which are all green edible materials. The cryogels exhibit rapid blood-triggered shape recovery and a high swelling ratio. The cryogels with 5 mg mL-1 HE (Gel-CS/OGA@HE5) exert excellent photothermal effects. Additionally, the cryogels have excellent cytocompatibility and blood clotting abilities. In the mouse liver injury model and mouse tail amputation model, Gel-CS/OGA@HE5 presents better hemostasis properties than gauze and a gelatin sponge. Moreover, Gel-CS/OGA@HE5 displays excellent healing performance as a wound dressing. Overall, we provide a simple and effective strategy to prepare cryogels for controlling wound bleeding and promoting wound healing.

Antibacterial aerogels with nano­silver reduced in situ by carboxymethyl cellulose for fresh meat preservation.

Bacterial cellulose (BC) was used as a reinforcing agent, citric acid (CA) as a cross-linking agent, and CMC@AgNPs as antibacterial nanomaterials, in which CMC@AgNPs were reduced from AgNO3 in situ by carboxymethyl cellulose (CMC) as a reducing agent and stabilizer to fight microbial corruption. Its potential application in packaging fresh meat has been investigated. Results showed that the antibacterial CMC@AgNPs/BC/CA aerogels with excellent structural integrity and outstanding water absorption were developed by adding 0.3% BC and 0.25% CA. The CMC@AgNPs/BC/CA aerogel significantly reduced the color change and the total viable bacterial counts (TVC) in fresh meat after 7 days of refrigerated storage. The results indicated that CMC@AgNPs/BC/CA aerogels can effectively extend the shelf life of fresh meat, and can be used for meat packaging as a biologically active absorption pad.

Chitosan­sodium alginate-collagen/gelatin three-dimensional edible scaffolds for building a structured model for cell cultured meat.

Cell cultured meat (CCM) production is an innovative technology that does not depend on livestock farming practices to produce meat. The construction of structured CCM requires a three-dimensional (3D) scaffold to mimic the extracellular matrix to provide mechanical support for the cells. Furthermore, the 3D scaffolds should be edible and have good biocompatibility and tissue-like texture. Here, we demonstrated a 3D edible chitosan­sodium alginate-collagen/gelatin (CS-SA-Col/Gel) scaffold that can support the adhesion and proliferation of porcine skeletal muscle satellite cells, culminating in the construction of a structured CCM model. The 3D edible scaffolds were prepared by freeze-drying using electrostatic interactions between chitosan and sodium alginate. Initially, the physicochemical properties and structural characteristics of different scaffolds were explored, and the biocompatibility of the scaffolds was evaluated using the C2C12 cell model. The results showed that the 2-CS-SA-Col1-Gel scaffold provided stable mechanical support and abundant adhesion sites for the cells. Subsequently, we inoculated porcine skeletal muscle satellite cells on the 2-CS-SA-Col1-Gel scaffold and induced differentiation for a total of 14 days. Immunofluorescence staining results showed cytoskeleton formation, and Western blotting (WB) and qPCR results showed upregulation of skeletal proteins and myogenic genes. Ultimately, the structured CCM model has similar textural properties (chewiness, springiness and resilience) and appearance to those of fresh pork. In conclusion, the method of constructing 3D edible scaffolds to prepare structured CCM models exhibits the potential to produce cell cultured meat.

Effects of quercetin on tenderness, apoptotic and autophagy signalling in chickens during post-mortem ageing.

The effect of quercetin on chicken breast muscle tenderness and the associated mechanism were investigated. The results indicated that quercetin significantly decreased the shear force and increased the myofibril fragmentation index (MFI). Haematoxylin-eosin-stained images showed that the internal structure of myofibril bundles in the quercetin-treated group was obviously degraded. Transmission electron microscopy showed that the myofibril structure, especially the M-line and A-band, was seriously degraded after quercetin treatment. Furthermore, quercetin treatment increased caspase-3 activity and the Bax/Bcl-2 ratio. The intensity of BiP, XBP1 and p-IRE1/IRE1 ratio increased significantly, and caspase-12 was activated. In addition, quercetin induced the transition from LC3I to LC3II and increased the expression of ATG7 and Beclin-1. The PI3K/Akt/mTOR signalling pathway was involved in the induction of autophagy and apoptosis by quercetin. These results indicated quercetin can promote meat tenderization, and activate apoptosis and autophagy pathways during post-mortem ageing.

Dual role (promotion and inhibition) of transglutaminase in mediating myofibrillar protein gelation under malondialdehyde-induced oxidative stress.

This study investigated the effects of transglutaminase (TGase) on the properties of myofibrillar protein (MP) and its heat-induced gels under malondialdehyde (MDA)-induced oxidation. The physicochemical characteristics, protein aggregation and rheological properties of MP were assessed. The gelling behaviours of MP were analysed with measurements of gel strength, cooking loss, microstructure and secondary structure. Under varying degrees of MDA oxidation, the addition of TGase always led to changes in the tertiary structure, loss of free amine and thiol groups, crosslinking of the myosin heavy chain, and decreasing solubility. However, the effect of TGase on MP gel quality differed. At 6 mmol/L MDA, the addition of TGase reduced the quality of MP gels by increasing cooking loss. However, at 12 mmol/L MDA, TGase reduced both the cooking loss and gel strength.

Effect of MTGase on silver carp myofibrillar protein gelation behavior after peroxidation induced by peroxyl radicals.

The objective of this study was to explore the oxidative modification induced by AAPH (2,2'-azobis (2-amidinopropane) dihydrochloride) on the microbial transglutaminase (MTGase) cross-linking reaction and gelling properties of silver carp myofibril protein (SCMP). Compared to AAPH treatment, MTGase addition resulted further changes of protein properties as evidenced by the decreasing free amino and thiol group content, the decreased secondary and tertiary structure, and the increasing storage modulus (G') and gel strength (P < 0.05). The proper oxidation induced by AAPH (5 mM) promoted the glutamine-lysine and disulfide cross-linking due to MTGase (10 U/g). Therefore, the quality of the SCMP gel was improved with a good water-holding capacity (WHC), gel strength and G'. This results could lay a theoretical foundation for the development of silver carp surimi products with good quality. Chemical compounds: (2,2'-azobis(2-amidinopropane)dihydrochloride (PubChem CID:76344); O-Phthalaldehyde (PubChem CID:4807); 5,5'-Dithiobis(2-Nitrobenzoic Acid) (PubChem CID:6254); 8-Anilino-1-naphthalenesulfonic acid (PubChem CID:1369); Acrylamide (PubChem CID: 6579); ß-Mercaptoethanol (PubChem CID: 1567); Sodium dodecyl sulfate (PubChem CID:3423265).

The gelation properties of myofibrillar proteins prepared with malondialdehyde and (-)-epigallocatechin-3-gallate.

Impact of malondialdehyde (MDA) and (-)-Epigallocatechin-3-gallate (EGCG) on gelling properties of myofibrillar proteins (MPs) was investigated. Addition of 6 mM MDA enhanced molecular interactions of proteins, thus the strength and elastic modulus (G') of gel were improved. EGCG addition aggravated gel quality deterioration due to further modification of MPs induced by EGCG. Addition of 12 mM MDA jeopardized gel quality according to the increasing of strength and G', but the decreasing of water-holding capacity (WHC), and the collapse of microstructure. Nevertheless, EGCG reacted with MDA forming EGCG-MDA adducts, hence improved gel quality, which was supported by the decreasing of strength, but the increasing of WHC, and the repaired microstructure of gel at 12 mM MDA. Addition of 24 mM MDA severely jeopardized gel quality, which became even worse due to EGCG addition. This work is helpful to understand the impact of MDA and polyphenols on the gel-forming capacity of MPs.

Active biodegradable films based on the whole potato peel incorporated with bacterial cellulose and curcumin.

Development of biodegradable food packaging using biomass based materials derived from agricultural wastes has been a trend in recent years. The biopolymer films were prepared using 3% and 5% (w/w) potato peel (PP) powder. Bacterial cellulose (BC) (0, 5, 10 and 15% based on PP powder) was added as a reinforcement agent. The scanning electron microscopy (SEM) revealed that 10% BC had a promising compatibility with the PP matrix. X-ray diffraction (XRD) and thermogravimetric analysis (TGA) showed that the crystallinity and the thermal stability of films did not change with BC addition. Fourier transform infrared spectroscopy (FTIR) indicated the hydrogen bonding interactions between the PP matrix and BC in the films. BC addition significantly improved the tensile strength (TS), but reduced their water vapor permeability (WVP), oxygen permeability (OP) and moisture content (MC) of the PP films. Addition of curcumin further increased the antioxidant properties of the PP films. The PP films with 1-5% curcumin significantly reduced lipid oxidation in the fresh pork during storage with lower malondialdehyde (MDA) content.