Myofibrillar Proteins Research Articles

Effect of bacterial fermentation on the ability of myofibrillar proteins to bind esters and its potential mechanism: Based on protein metabolism and structural changes

The effect of fermentation strains (Lactiplantibacillus plantarum CQ01107 and Staphylococcus simulans CD207) on the binding properties of porcine myofibrillar proteins (MPs) to esters was investigated from two perspectives: metabolism degree and structural alterations. Results demonstrated that S. simulans could reduce the particle size and α-helix content of MPs, while simultaneously increasing the absolute zeta potential and active sulfhydryl content. This process decreased protein aggregation and facilitated the unfolding of MPs, thereby enhancing their binding to esters. Conversely, L. plantarum showed limited promotion, which might be related to its robust acid production and protein hydrolysis capacities. In addition, ethyl octanoate, with a longer carbon chain length, was found to have the highest binding capacity to MPs (28.38 %–41.59 %). Molecular docking results further revealed that the binding of the four esters to MPs was spontaneous, with ethyl octanoate exhibiting the lowest binding energy to MPs (−5.635 kcal/mol). The primary forces involved in the binding of the four selected esters to MPs were hydrophobic interactions, hydrogen bonding, and van der Waals forces. These findings can provide new insights into the mechanisms by which fermentation strains influence flavor formation in fermented foods.

Effect of NaCl on the structure and digestive properties of heat-treated myofibrillar proteins

During meat processing, the quality of food and structure of meat proteins are affected by different processing technologies and addition of raw and auxiliary materials. Different meat products are treated with varying processing temperatures and NaCl content, changing the protein molecular structure. This study aimed to determine impact of heating temperature (40–115 °C) and NaCl concentration (0–0.8 M) on the oxidation, structure, and digestibility of beef myofibrillar proteins. The results revealed that high temperatures and NaCl concentration of 0.4–0.8 M caused the salting-out effect, leading to a decrease in solubility. The oxidative denaturation of proteins leads to increased protein aggregation. Consequently, structural changes of myofibrillar proteins, and the digestive enzymes are unable to recognize specific sites, which reduces the digestibility of the proteins. The findings of this study revealed that heating beef myofibrillar proteins at 85 °C and 0.4 M NaCl substantially improved its digestibility to 85.66 %.

Exploring the potential of fibrinogen hydrolysates as enhancers for myofibrillar protein gelation: Insights into molecular assembly behavior

This study explored the use of pig blood fibrinogen hydrolysates, enzymatically hydrolyzed with trypsin and flavorzyme, to enhance myofibrillar protein gels, addressing issues like poor gel strength and water loss in meat products. By incorporating varying concentrations of fibrinogen hydrolysates into myofibrillar proteins, heat-induced gels were prepared. The composite gels showed improved textural properties, rheological characteristics and water-holding capacity. Scanning electron microscopy and atomic force microscopy analyses revealed a uniform, dense surface and an orderly internal structure in the composite gels. The study also noted decreased α-helix and random coil and increased β-sheet and β-turn contents, indicating a more ordered secondary structure. Hydrophobic interactions and disulfide bonds were identified as key factors in enhancing gelation, and a model was proposed to explain these molecular effects. This research demonstrates a potential of fibrinogen hydrolysates to improve quality and structure of myofibrillar protein gels designed for high-quality meat products.

Study the stability mechanism of Nano freezing-thawing on myofibrillar proteins in Atlantic salmon

Study the stability mechanism of Nano freezing-thawing on myofibrillar proteins in Atlantic salmon

Influence of low voltage programmed thawing on the quality of frozen pork and its myofibrillar protein

This study explored the effects of low voltage programmed thawing (LVPT) on thawing period, water status, current density as well as physicochemical properties of porcine longissimus dorsi muscle, considering air thawing (AT), water immersion thawing (WT) and cold storage thawing (CST) for the comparison, additionally making fresh meat (FM) as the control. The thawing process used current limiting to gradually reduce the maximum output current to 500, 200 and 100 mA, respectively, by applying a safe voltage of 35 V in a home defrosting apparatus. Using two pairs of foil plate, the thawing (LVPT-2) with dual current loop accelerated the melting of ice crystals than the process with one pair of polar plate (LVPT-1) with one current loop. From −18 °C to −1 °C, the thawing period of LVPT-2 was reduced by 14.54 %, 11.72 %, and 15.95 % compared with that of LVPT-1 for samples within the thickness of 2, 3 and 4 cm, respectively. During the process, the intensity and density of the current loaded on the frozen meat gradually increased. LVPT-1 retained water and reduced its migration inside the meat, and protein solubility was better maintained. As the thickness increased, the effect of LVPT-2 on the moisture distribution and protein solubility of the sample decreased. LVPT-1 treatment resulted in less fat oxidation as well as stable secondary and tertiary structures of myofibrillar proteins compared to other treatments. At low voltage, the ability of LVPT −2 to maintain meat quality was reduced due to the increase in current density. In addition, LVPT-1 reduced muscle damage because of rapid melting of ice crystals. In contrast, LVPT-2 was conductive the formation of hot spots due to dual current loop passed through the pork in final phase of thawing, which would adversely influence the quality of thawed meat. Finally, LVPT could improve the rate of thawing, water binding capacity and meat quality, especially, there was no overheating effect on the meat because of current limiting.

Having their cake and eat it too: Effects of different cations in reduced-salt myofibrillar protein microgel pickering emulsion under high-intensity ultrasound

Health concerns have increased the demand for low-salt emulsified meat products emulsions; however, salt-soluble myofibrillar proteins (MP) are less than optimal emulsifiers under low-salt conditions. In this study, four different cations (Na+, K+, Ca2+, and Mg2+) were added to a Pickering emulsion and high-intensity ultrasound (HIU, 20 kHz, 600 W) was used to determine the quality of the final emulsion. The divalent cation group was found to significantly (P < 0.05) improve the emulsification performance of the emulsion; in particular, the emulsion activity index and emulsion stability index of Mg2+-H group increased to 16.24 ± 1.22 m2/g and 89.66 ± 3.01%, respectively. Measurements of the particle size and zeta potential confirmed that the emulsion particle size of the Mg2+-H group decreased, and the electrostatic repulsion force increased. Based on the surface hydrophobicity and endogenous tryptophan spectra, the particle structure of the MP microgel was determined to be a more stable emulsifier. What's more, the divalent cations promoted the droplets to become evenly dispersed under high-intensity ultrasound. The micrograph confirmed this result: the divalent cation group emulsion was more uniform and stable than the monovalent cation group emulsion. Divalent cations are more suitable than monovalent cations for improving the quality of reduced-salt Pickering emulsions.

Recovery of protein-rich biomass from surimi rinsing wastewater by using a sustainable cold plasma treatment

Surimi rinsing wastewater is typically discarded, causing waste of protein resources and environmental pollution. This study investigated the technology of a cold atmospheric plasma jet (CAPJ) for the recovery of protein-rich biomass (PRB), including myofibrillar proteins (MPs) and sarcoplasmic proteins (SPs), from surimi rinsing wastewater. The protein recovery yield was up to 59.84 %. CAPJ induced an increase in carbonyl and decreased sulfhydryl in protein content. Furthermore, the secondary structure of the protein was unfolded, particularly the transition from α-helix to β-sheet. The formation of disulfide bonds and increased hydrophobic interactions promoted protein aggregation (the particle size from 185.76 nm to 1869.07 nm, P < 0.05) and reduced solubility. The proteomic results indicated that CAPJ increased the expression level of antioxidant enzymes. Overall, the CAPJ technology could recover proteins from surimi rinsing wastewater for industrial application, which will promote the sustainable development of the surimi industry.

Muscle Proteins, Technological Properties, and Free Amino Acids of Epaxial Muscle Collected from Asian Seabass (Lates calcarifer) at Different Postmortem Durations

The aim of this study was to compare the changes in the epaxial muscle proteins of Asian seabass at two different postmortem durations. The epaxial muscles of Asian seabass were collected 1 h or 24 h postmortem (PM). Whole, ungutted fish were stored in an ice box, with the ice refilled every two hours. The results show significant increases in the MFI values and the contents of solubilized sarcoplasmic proteins, with a molecular weight of proteins of 47 kDa in the 24 h PM samples (p &lt; 0.05). Myofibrillar and alkaline-soluble proteins in the epaxial muscle remained intact 24 h postmortem. Compared with the 1 h PM samples, the 24 h PM meat exhibited lower degrees of expressible water and hardness (p &lt; 0.05), indicating superior water-holding capacity and meat tenderness. However, no differences in springiness or cohesiveness of the cooked meat were observed. Free L-glutamic acid, known as an umami-tasting amino acid, was significantly increased upon the extension of postmortem duration, and its level was above the taste threshold concentration. Overall, the findings indicated that the 24 h PM epaxial muscle of Asian seabass exhibited superior technological properties, along with higher contents of taste-related amino acids.

Improving gel properties of low-salt silver carp surimi through single-mode microwave-assisted processing

Surimi, a product derived from low-value fish, is widely used to produce high-value items such as imitation crabsticks and fish balls. However, surimi from freshwater fish like silver carp generally exhibits inferior gel properties compared to marine fish. This study investigated the effects of 915 MHz single-mode microwave-assisted processing on the gel properties of low-salt silver carp surimi. Traditional water bath processing served as the control for comparison. Gel strength, texture, water-holding capacity (WHC), whiteness, protein secondary structure, and microstructure, along with thermal processing efficiency for microbial inactivation were evaluated. The results demonstrated that single-mode microwave processing significantly improved gel properties compared to water bath processing. Compared with the optimal water bath treatment, the gel strength of microwave processed samples increased significantly from 479.32 g·cm to 821.74 g·cm, while WHC increased from 0.77 to 0.87. The microwave processed surimi exhibited a more uniform microstructure. Because it enhanced the dissolution and cross-linking of myofibrillar proteins, leading to superior gel formation characterized by a lower α-helix content (10.31%) and higher β-sheet content (61.70%). Furthermore, microwave processing achieved faster heating rates, reducing the time spent in the critical gel deterioration temperature range, thereby improving overall gel quality. Single-mode microwave-assisted gelation also provided better thermal processing uniformity and efficiency, achieving pasteurization standards. The study highlights the potential of 915 MHz single-mode microwave processing to produce high-quality, low-salt surimi products, meeting contemporary health and convenience demands. These findings offer valuable insights for the industrial application of single-mode microwave technology in surimi production, promoting the development of innovative processing techniques to enhance the quality of surimi-based food products.

Antioxidant potential and stability of bovine liver peptides in fermented sausage (sucuk): A pathway to value-added functional meat products

The objective of the present study was to evaluate the stability of low-molecular-weight peptides after their incorporation into fermented sausages. Peptide fractions were produced from the bovine liver peptides (BLP) by enzymatic hydrolysis. Optimal conditions were identified as hydrolysis at 45 °C for 24 h, resulting in the generation of <3 kDa and <10 kDa peptide fractions exhibiting the highest antioxidant activity. The antioxidant potential of fermented sausages made from beef meat containing BLPs at 0.5% and 1% concentrations was assessed during manufacturing and storage. The initial antioxidant activity values of the sausages containing BLPs were higher than that of the control. Furthermore, the stability of the peptides in the sausages was demonstrated by their preserved antioxidant activity for 102 days. Peptides extracted from 1% of <10 kDa-peptide containing sausages possessed two-fold higher radical scavenging activity than control. The highest peptide concentration in the sausages containing 1% of <10 kDa-peptides (79.32±1.34–101.56±1.22 μg peptide/mg) was maintained from day 0 to day 102. The SDS-PAGE profile of myofibrillar proteins and peptide extracts revealed the degradation of proteins were delayed in <10 kDa of peptide added sausages, suggesting that the protein oxidation was slowed down. These results contribute to the advancement of peptide commercialization as functional ingredients in fermented food products. Furthermore, our research sheds light on the enduring impact of fermentation on the stability of the added peptides and the synergistic interactions between naturally produced peptides during fermentation and those introduced externally.

Proteomic analysis of wild boar meat: Effect of storage method and time on muscle protein stability

Oxidation processes affect proteins from various molecular pathways and are crucial for wild boar meat quality, shelf life and human health. This study investigated the effects of different storage methods on the formation and composition of oxygen-induced protein aggregates in the muscles of European wild boar (Sus scrofa scrofa). Vacuum packaging (VAC), modified atmosphere packaging (MAP) and dry-ageing (DA) were compared over a 21-day storage period. The results showed significant differences in protein aggregation depending on the method and storage time. The most intense protein aggregation occurred in the MAP (80 % O2), while air DA (20.9 % O2) resulted in intermediate levels of protein aggregation. Crucial myofibrillar proteins involved in aggregate formation were titin, myosin isoforms (MYH1, MYH2 and MYH7) and nebulin, which were cross-linked with small sarcoplasmic enzymes, such as muscle creatine kinase, isocitrate dehydrogenase and ATPase 1. High‑oxygen storage conditions also promoted the oxidation of ATP synthase, beta-enolase 3, ADP/ATP translocase and myoglobin.

Differentiated texture and lipid-lowering effect of cooked pork belly by different cooking regimes: insights from myofibrillar protein aggregation and dissociation, micromorphology

The effects of traditional flame stewing (TFS), commercial pressure cooking (PC), and sous-vide cooking (SVC) on the texture of pork belly and the underlying mechanism were investigated. SVC treatment at a low temperature (60 °C) achieved weaker oxidation of sulfhydryl groups, lower α-helical loss, higher surface hydrophobicity, and actomyosin dissociation, which reduced the shrinkage of myofibrils and loss of immobilized water, thus contributed to a tender meat. However, the protein aggregation predominated without actomyosin dissociation during PC treatment, causing the toughening of meat. TFS and PC intensified the atrophy of adipocytes and damage of cell membranes, which helped for the softening of adipose tissue and achieved 13.82% and 17.62 % lipid loss, respectively, and contributed to a higher ratio of polyunsaturated fatty acids to saturated fatty acids (0.45–0.46) than SVC (0.34). Comparisons showed that TFS was a more suitable method to prepare cooked pork belly with tender and juicy texture, low lipid content, and high ratio of polyunsaturated fatty acids to saturated fatty acids. This study offered new insights into how cooking regimes affect the texture and lipid-lowering effects of stewed pork.

Proteomic analysis revealing mechanisms of κ-carrageenan modulating the gelling properties of heat-induced microbial transglutaminase cross-linked myofibrillar protein gels

The effects of κ-carrageenan (KC, 0.2 %, w/w) on the gelling properties and microstructures of heat-treated myofibrillar protein (MP) gels mediated by various levels (0, 0.1, 0.3, and 0.5 %, w/w) of microbial transglutaminase (TG) were investigated. The results revealed that regardless of TG level, the existence of KC significantly promoted the forming of disulphide bonds but limited the generation of large-sized protein polymers in MP gel treated with TG alone, as well as subsequently enhanced the water retention ability and gel strength of mixed MP gel, especially in the sample adding 0.3% TG and 0.2% KC, which was clearly verified by the forming of more denser and continuous gel network. Moreover, TG-induced changes in protein structure led to the alteration in the protein profiles of different MP gels, and an obvious down-regulation trend of myosin (Q9TV63, Q9TV62, P79293, and Q9TV61) was identified in the TG-0.3 group. Furthermore, liquid chromatography–tandem mass spectrometry analysis demonstrated that TG mainly induces protein crosslinking in the light meromyosin domain of the myosin rod. Meanwhile, in the presence of KC, the crosslinking-site percentage of the TG-0.3 group decreased from 37.7% to 36.2%, indicating that KC potentially reduces TG-induced crosslinking in MP gels. Thus, our findings illustrate the critical mechanisms underlying the modulation of KC on the gelling properties of TG-mediated MP gels at the molecular level, which providing potential guideline for the combined applications of TG and KC in the meat industry.

Influence of droplet size and surface hydrophobicity of soybean protein-based nanoemulsion fillers on the quality of silver carp myofibrillar protein gels

This study investigated the mechanisms underlying the influence of droplet size and emulsifier wettability on gel properties when oil-in-water (O/W) emulsions serve as fillers in myofibrillar protein (MP) gels. Pickering emulsions with varying droplet sizes were prepared using soybean protein isolate (SPI) and SPI-curcumin nanoparticles, then used to construct composite gels. Findings showed that decreased droplet size and increased emulsifier surface hydrophobicity enhanced hydrophobic interactions in the gel, increasing the β-sheet content of MP molecules. Upon the introduction of SPI-Cur-NPs stabilized nanoemulsion (SCNE), the hydrophobic force in the gel was approximately 2.6-fold more remarkable than that of the control, and the β-sheet content increased to 16.51 %. This resulted in a denser mesh framework and more uniform oil droplet distribution, increasing the hardness value from 26.993 g to 41.847 g. Moreover, SCNE addition improved gel antioxidant properties, reducing carbonyl and peroxide levels to 31.82 % and 24.15 % of the control, respectively. These findings offer insights for improving MP-based gel products.

Effect of chitosan on thermal gelling properties of pork myofibrillar protein and its mechanism.

Previous studies have demonstrated that the addition of chitosan can improve the quality and functional properties of meat products. However, the underlying mechanism remains unclear. In this study, the effect and mechanism of the addition of chitosan on the gel properties of myofibrillar protein (MP) were investigated. The results indicated that the gel strength and the water-holding capacity of MP-chitosan gel increased significantly when chitosan was added at 2.5-10 mg mL-1. Myofibrillar protein samples with 10 mg mL-1 added chitosan exhibited the highest elasticity and viscosity during gel formation and strengthening. The addition of chitosan also caused a modification in both the secondary and tertiary structure of MP, resulting in an enhanced exposure of hydrophobic and sulfhydryl groups in comparison with the control. Chitosan inhibited the conversion of immobilized water into free water and the formation of water channels during the thermal gelation process of MP. The denaturation enthalpy (ΔH) of myosin decreased as the concentration of chitosan exceeded 5 mg mL-1. The microstructure showed that the incorporation of chitosan (5-10 mg mL-1) facilitated the formation of compact and well organized MP gel networks. The addition of chitosan can enhance the functional properties of meat protein and facilitate heat-induced gelation, making it a promising ingredient for improving the quality of processed meat products. © 2024 Society of Chemical Industry.

Cyclic Continuous Glycation Enhanced Dispersibility of Myofibrillar Protein: Reaction Efficiency and Sites Modification.

Reaction efficiency in glycation lacks sufficient attention, leading to the waste of process costs. Cyclic continuous glycation (CCG) is an effective approach to accelerate covalent binding between myofibrillar protein (MP) and glucose. This study elucidated that CCG promoted the exposure of reactive glycated sites in MP with full unfolding of secondary and tertiary structures. Notably, the glycation rate was significantly increased by 65.43%. Physicochemical properties indicated that MP-glucose conjugates with high graft degree exhibited favorable solubility, dispersibility, and thermal stability. Furthermore, proteomics was applied to reveal the glycated sites and products in glycoconjugates of MP. Glycation preferentially acted on the tails of the myosin heavy chain. The glucosylation modification on the head region was enhanced by CCG contributing to the inhibition of the head-head interaction. Overall, this study systematically clarifies the mechanism of CCG, providing a theoretical basis for the application of glycation in innovative meat products.

Impact of lipid oxidation products on the digestibility and structural integrity of Myofibrillar proteins during thermal processing

This study explores the effects of lipid oxidation products (LOPs), specifically CHP, t,t-DDE, and MDA, on the digestibility and structural integrity of myofibrillar proteins (MP) during processing. LOPs were first assessed by heating at 180 °C for 15 min, showing a significant reduction in digestibility in MDA-treated samples (65.40 %), followed by t,t-DDE (45.10 %) and CHP (13.07 %). MALDI-TOF MS analysis revealed decreased peptide abundance and lower average molecular weight in t,t-DDE- and MDA-treated samples. Notably, substantial decreases in α-helix content and increases in random coil structures were detected, particularly in MDA-treated samples. Assessments of surface hydrophobicity and thiol content underscored the detrimental impact of secondary LOPs on MP structure. Higher MDA concentrations led to a substantial reduction in intrinsic fluorescence intensity, along with an increase in Schiff base content. A PLS regression model demonstrated strong predictive capabilities for MP digestibility, highlighting the importance of optimizing meat processing parameters to minimize nutritional degradation.

Effect of different precooling methods on the physicochemical changes, water migration and myofibrillar protein characteristics of pufferfish (Takifugu obscurus) during cold storage

SummaryThe objective of this study was to evaluate the effect of slurry ice (SI) in physicochemical changes, water migration, myofibrillar protein characterisation, and extended shelf life during cold storage of pufferfish (Takifugu obscurus). SI provided faster cooling rate, better structural stability of muscle tissue, and a significant reduction in total viable count (TVC). In addition, the use of SI treatment resulted in a considerable reduction in total volatile basic nitrogen (TVB‐N) and pH. The results of the texture profile analysis (TPA) demonstrated that SI had a significant inhibitory effect on the reduction of hardness, springiness, resilience, and chewiness. In addition, SI treatment better maintained a well‐structured myofibrillar protein stabilising the tertiary structure. Low‐field nuclear magnetic resonance (LF‐NMR) and magnetic resonance imaging (MRI) revealed that SI treatment could retard the water migration. Furthermore, the results of confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) indicated that SI treatment of pufferfish effectively slowed down the degradation of their myofibrillar protein fraction throughout the cold storage.

Contribution of ultrasound‐assisted protein structural changes in marinated beef to the improved binding ability of spices and flavor enhancement

AbstractBACKGROUNDMarination is an important part of air‐dried beef processing, and traditional methods are inefficient and produce inconsistent results. Ultrasound, as a novel technology, can be combined with traditional marination methods. The study aimed to investigate the improvement of beef flavor by ultrasound‐assisted marination. At the same time, the potential relationship between the alteration of meat protein and flavor quality by ultrasound‐assisted marinating was further investigated to enable better flavor modulation and research.RESULTSHeadspace solid‐phase microextraction–gas chromatography–mass spectrometry revealed that the spice flavor of beef was significantly enhanced by 500 W ultrasound‐assisted marination. Meanwhile, the experimental results demonstrated that the ultrasound‐assisted marination promoted the unfolding of beef myofibrillar protein structure, which increased the number of hydrophobic and hydrogen bonding sites, enhanced the electrostatic effect and improved the functional properties of the protein. Ultrasound‐assisted marination significantly enhanced the binding ability of beef myofibrillar proteins to flavor compounds compared with conventional marination. An electronic nose confirmed that this resulted in a significant increase in the flavor of the marinated meat.CONCLUSIONUltrasound‐assisted marination effectively enhanced the flavor of marinated meat, which was closely related to the development of protein conformation. The results of this study have important implications for the food industry and the role of protein unfolding processes in flavor modulation. In particular, the findings can be practically applied to improving meat flavor under ultrasound‐assisted marination. © 2024 Society of Chemical Industry.

Exploring the Effects of S-Nitrosylation on Caspase-3 Modification and Myofibril Degradation of Beef In Vitro.

This study aimed to explore the effects of S-nitrosylation on caspase-3 modification and its subsequent effects on beef myofibril degradation in vitro. Recombinant caspase-3 was reacted with different concentrations of S-nitrosoglutathione (GSNO, nitric oxide donor) at 37 °C for 30 min and subsequently incubated with purified myofibrillar protein from bovine semimembranosus muscle. Results indicated that the activity of caspase-3 was significantly reduced after GSNO treatments (P < 0.05) and showed a dose-dependent inhibitory effect, which was attributed to the increased S-nitrosylation extent of caspase-3. LC-MS/MS analysis revealed that caspase-3 was S-nitrosylated at cysteine sites 116, 170, 184, 220, and 264. Moreover, the degradation of desmin and troponin-T was notably suppressed by S-nitrosylated caspase-3 (P < 0.05). To conclude, protein S-nitrosylation could modify the cysteine residues of caspase-3, which accounts for the reduced caspase-3 activity and further represses its proteolytic ability on beef myofibrillar protein.