Investigating the effects of low-salt processing on the umami peptides of dry-cured ham using peptidomics techniques.

This study investigated the effect of low-salt processing on the umami peptide profile of dry-cured hams. Peptidomics data showed 633 umami peptides in the low- and full-salt groups. Among them, 36.2% and 26.5% of shared umami peptides in the low-salt group were significantly down- and up-regulated in relative abundance. Multivariate statistical analysis showed 1011 significantly different umami peptides (SDUPs) in the low- and full-salt groups. Creatine kinase M-type (CKM) and fast skeletal muscle troponin T (TnTf) were the main precursor proteins of these SDUPs. At the end of processing, the relative expression of CKM was lower in the low-salt group than in the full-salt group (P < 0.05), but there was no significant difference in TnTf. More dipeptidyl peptidase cleavage sites were observed in CKM and TnTf proteins in the low-salt group.

Effects of Long-Term Administration of Bovine Bone Gelatin Peptides on Myocardial Hypertrophy in Spontaneously Hypertensive Rats.

The research purpose was to investigate the effects and the underlying molecular mechanisms of bovine bone gelatin peptides (BGP) on myocardial hypertrophy in spontaneously hypertensive rats (SHR). BGP relieved myocardial hypertrophy and fibrosis in SHR rats in a dose-dependent manner by reducing the left ventricular mass index, myocardial cell diameter, myocardial fibrosis area, and levels of myocardial hypertrophy markers (atrial natriuretic and brain natriuretic peptide). Label-free quantitative proteomics analysis showed that long-term administration of BGP changed the left ventricle proteomes of SHR. The 37 differentially expressed proteins in the high-dose BGP group participated in multiple signaling pathways associated with cardiac hypertrophy and fibrosis indicating that BGP could play a cardioprotective effect on SHR rats by targeting multiple signaling pathways. Further validation experiments showed that a high dose of BGP inhibited the expression of phosphoinositide 3-kinase (Pi3k), phosphorylated protein kinase B (p-Akt), and transforming growth factor-beta 1 (TGF-ß1) in the myocardial tissue of SHR rats. Together, BGP could be an effective candidate for functional nutritional supplements to inhibit myocardial hypertrophy and fibrosis by negatively regulating the TGF-ß1 and Pi3k/Akt signaling pathways.

Study on the anti-inflammatory activity of the porcine bone collagen peptides prepared by ultrasound-assisted enzymatic hydrolysis.

In this study, the effects of ultrasound-assisted enzymatic hydrolysis on the extraction of anti-inflammatory peptides from porcine bone collagen were investigated. The results showed that ultrasound treatment increased the content of α-helix while decreased ß-chain and random coil, promoted generation of small molecular peptides. Ultrasound-assisted enzymatic hydrolysis improved the peptide content, enhanced ABTS+ radical scavenging and ferrous ion chelating ability than non-ultrasound group. At the ultrasonic power of 450 W (20 min), peptides possessed significant anti-inflammatory activity, where the releasing of interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) was all suppressed in lipopolysaccharide (LPS) induced RAW264.7 cells. After the analysis with LC-MS/MS, eight peptides with potential anti-inflammatory activities were selected by the PeptideRanker and molecular docking. In general, the ultrasound-assisted enzymatic hydrolysis was an effective strategy to extract the bioactive peptides from porcine bone, and the inflammatory regulation capacity of bone collagen sourced peptides was firstly demonstrated.

Effect of Yeast-Derived Peptides on Skeletal Muscle Function and Exercise-Induced Fatigue in C2C12 Myotube Cells and ICR Mice.

In our previous study, the antioxidant peptides (XHY69AP, AP-D, YPLP, and AGPL) were obtained from potential probiotic yeast (Yamadazyma triangularis XHY69), which was selected by our lab from dry-cured ham. This work aimed to explore the effects of yeast-derived peptides on skeletal muscle function and muscle fatigue. Results showed that yeast-derived peptides up-regulated slow-twitch fiber expression and down-regulated fast-twitch fiber expression in C2C12 cells (p < 0.05). The peptides improved mitochondrial membrane potential, adenosine triphosphate generation, and expression of cytochrome-relative genes, thus promoting mitochondrial function. Among these peptides, YPLP up-regulated the relative gene expression of the AMP-activated protein kinase (AMPK) pathway and activated AMPK by phosphorylation. Moreover, YPLP could prolong treadmill time, increase muscle and liver glycogen contents, reduce lactic acid and urea nitrogen contents, and alleviate muscle tissue injury in ICR exercise mice. These results demonstrate that yeast-derived peptides could change the muscle fiber composition, improve muscle function, and relieve muscle fatigue.

Effects of ultrasound on the oxidation and structure of myofibrillar protein in the presence or absence of soybean oil.

BACKGROUND: Ultrasound is widely used as a novel non-thermal processing technique to improve protein properties. In recent decades, applying ultrasound-assisted emulsification (UAE) to produce protein-stabilized emulsion has attracted people's attention. Instead of applying ultrasound to treat a single protein solution, UAE treatment refers to the use of sonication to a mixture of protein and oil. The purpose of this study was to compare the different effects of ultrasound treatment on the properties of myofibrillar protein (MP) in the presence or absence of soybean oil. A suitable sonication power was selected based on the change in emulsion properties. RESULTS: 300 W sonication power was selected because of its most effectively decreased emulsion droplet size and increased absolute zeta potential. Sonication more significantly increased the protein carbonyl content and disulfide bonds of the MP-soybean oil sample compared with the MP sample. Due to the presence of oil, ultrasound could unfold more protein molecules, illustrated by a lower α-helix content and intrinsic fluorescence intensity, and a higher surface hydrophobicity. Results of liquid chromatography-tandem mass spectrometry illustrated that sonication enhanced the myosin heavy chain and actin content at the soybean oil interface as well as accelerated the myosin light chain to separate from myosin in the MP-soybean oil system. CONCLUSION: Ultrasound treatment could lead to a higher level of protein oxidation and greater protein molecule exposure in the MP in the presence of oil system than in the oil-free MP system. © 2023 Society of Chemical Industry.

Effects of carboxymethyl cellulose on the emulsifying, gel and digestive properties of myofibrillar protein-soybean oil emulsion.

Improving the qualities of vegetable oil replaced animal fat meat products is particularly fascinating for the development of healthy meat products. This work was designed to investigate the effects of different carboxymethyl cellulose (CMC) concentrations (0.01 %, 0.05 %, 0.1 %, 0.2 %, and 0.5 %) on the emulsifying, gelation, and digestive properties of myofibrillar protein (MP)-soybean oil emulsions. The changes in MP emulsion characteristics, gelation properties, protein digestibility, and oil release rate were determined. Results demonstrated that CMC addition decreased the average droplet size and increased the apparent viscosity, storage modulus, and loss modulus of MP emulsions, and a 0.5 % CMC addition significantly increased the storage stability during 6 weeks. Lower CMC addition (0.01 % to 0.1 %) increased the hardness, chewiness, and gumminess of emulsion gel especially for the 0.1 % CMC addition, while higher CMC (0.5 %) content decreased the texture properties and water holding capacity of emulsion gels. The addition of CMC decreased protein digestibility during the gastric stage, and 0.01 % and 0.05 % CMC addition significantly decreased the free fatty acid release rate. In summary, the addition of CMC could improve the stability of MP emulsion and the texture properties of the emulsion gels, and decrease protein digestibility during the gastric stage.

Moderate Protein Oxidation Improves Bovine Myofibril Digestibility by Releasing Peptides in the S2 Region of Myosin: A Peptidomics Perspective.

This study aimed to investigate the influence of protein oxidation on the digestive properties of beef myofibrillar protein (MP). MP was treated with a hydroxyl radical-generating system containing various concentrations of H2O2. The increased content in a free sulfhydryl group and surface hydrophobicity indicated that oxidation treatment with 1 mM H2O2 induced unfolding of MP. Reducing and nonreducing SDS-PAGE results suggested that 10 mM H2O2 oxidation treatment resulted in aggregation of MP; meanwhile, the disulfide bond was the major covalent bond involved in aggregation. Peptidomics showed that peptides in the digestion products of MP were mainly derived from myosin tail. Moderate oxidation (1 mM H2O2) facilitated the release of peptide in the rod portion (S2) of myosin, whereas excessive oxidation (10 mM H2O2) inhibited peptide release in the light meromyosin region. This work presents insightful information for the crucial impact of oxidation on meat protein digestibility from the peptidomics perspective.

Bovine Bone Gelatin-Derived Peptides: Food Processing Characteristics and Evaluation of Antihypertensive and Antihyperlipidemic Activities.

This study aimed to evaluate the food processing properties of bovine bone gelatin-derived peptides (BGPs) and their effects and mechanisms on hypertension and hypertension complications in spontaneously hypertensive rats (SHRs). BGPs had good acid, high temperature, and NaCl resistance abilities in vitro. Additionally, Maillard reaction of BGPs with low-dose reducing sugar (≤15%) exhibited a free radical scavenging effect. BGPs significantly reduced the blood pressure, triglyceride levels, and the low-density lipoprotein cholesterol/high-density lipoprotein cholesterol ratio in SHRs through downregulated angiotensin converting enzyme (ACE), angiotensin II (Ang II), and Ang II type 1 receptor (AT1R) levels and the upregulated Ang II type 2 receptor (AT2R) level. In brief, BGP could alleviate hypertension and dyslipidemia in SHRs by inhibiting ACE/Ang II/AT1R and activating the Ang II/AT2R signaling pathway. Our study suggests that BGP has good food processing properties and could be a potential nutraceutical for antihypertensive and antihyperlipidemic issues.

The Anti-Inflammatory Effect of Bovine Bone-Gelatin-Derived Peptides in LPS-Induced RAW264.7 Macrophages Cells and Dextran Sulfate Sodium-Induced C57BL/6 Mice.

The bioactive peptides hydrolyzed from bone collagen have been found to possess health-promoting effects by regulating chronic diseases such as arthritis and hypertension. In the current study, the anti-inflammatory effect of bovine bone gelatin peptides (GP) was evaluated in 264.7 macrophages cells and followed by animal trials to investigate their interference on inflammatory cytokines and gut microbiota compositions in dextran sodium sulfate (DSS)-induced C57BL/6 mice. The GP was demonstrated to alleviate the extra secretion of interleukin-6 (IL-6), nitric oxide (NO) and tumor necrosis factor-α(TNF-α) in lipopolysaccharide (LPS)-induced RAW264.7 cells. In DSS-induced colitis mice, the gavage of GP was demonstrated to ameliorate the IBD symptoms of weight loss, hematochezia and inflammatory infiltration in intestinal tissues. In serum, the proinflammatory cytokines (TNF-α,IL-6, MCP-1, IL-1ß) were suppressed along with the decreasing effect on toll-like receptor 4 and cyclooxygenase-2 by GP treatment. In the analysis of gut microbiota, the GP was checked to modulate the abundance of Akkermansia, Parasutterella, Peptococcus, Bifidobacterium and Saccharibacteria. The above results imply that GP could attenuate DSS-induced colitis by suppressing the inflammatory cytokines and regulating the gut microbiota.

Marine Products As a Promising Resource of Bioactive Peptides: Update of Extraction Strategies and Their Physiological Regulatory Effects.

Marine products are a rich source of nutritional components and play important roles in promoting human health. Fish, mollusks, shellfish, as well as seaweeds are the major components of marine products with high-quality proteins. During the last several decades, bioactive peptides from marine products have gained much attention due to their diverse biological properties including antioxidant, antihypertensive, antimicrobial, antidiabetic, immunoregulation, and antifatigue. The structural characteristics of marine bioactive peptides largely determine the differences in signaling pathways that can be involved, which is also an internal mechanism to exert various physiological regulatory activities. In addition, the marine bioactive peptides may be used as ingredients in food or nutritional supplements with the function of treating or alleviating chronic diseases. This review presents an update of marine bioactive peptides with the highlights on the novel producing technologies, the physiological effects, as well as their regulation mechanisms. Challenges and problems are also discussed in this review to provide some potential directions for future research.

The physiological activity of bioactive peptides obtained from meat and meat by-products.

Meat and meat products constitute an important source of nutrients and play vital roles for growth, maintenance and repair of the body. In addition to the high quality of proteins, meat is also regarded as a major resource to produce bioactive peptides. Meat processing industry also produces by-products such as bones, blood and viscera, which could be further used for the production of bioactive compounds. In the physiological analysis, meat bioactive peptides have been reported to exert antioxidant, anti-hypertensive, anti-inflammatory, anti-microbial and antitumoral activities, which endow nutritional and functional value of meat. With the objective to exert the functional effect, the bioavailability should also be considered due to the degradation by digestion enzymes and the absorption process in intestinal mucosa. In this chapter, the general source, the enzymatic hydrolysis, the physiological effects as well as the bioavailability of bioactive peptides in meat are discussed.

Proteomics identification of differential S-nitrosylated proteins between the beef with intermediate and high ultimate pH using isobaric iodoTMT switch assay.

This study aimed to identify the differential S-nitrosylated proteins at post-mortem in beef longissimus thoracis (LT) muscles from carcasses with intermediate (5.40 < pH < 5.80, n = 6) and high (pH ≥ 6.00, n = 6) ultimate pH (pHu). LT muscles were labeled with iodo-tandem mass tags (iodoTMT126-129). A total of 856 S-nitrosylated sites from 257 proteins were identified in high pHu beef LT muscles. The S-nitrosylated protein intensity in high pHu beef was higher compared to that of intermediate pHu beef along with a large number of cysteine sites. The motif revealed that the cysteine modifications played an essential role in cellular signaling and homeostasis. In high pHu beef, more up-regulated proteins were related to energy metabolism enzymes and mitochondrial dysfunction enzymes compared to down-regulated proteins which are involved in calcium homeostasis.

γ-Glutamylvaline Prevents Low-Grade Chronic Inflammation via Activation of a Calcium-Sensing Receptor Pathway in 3T3-L1Mouse Adipocytes.

The calcium-sensing receptor (CaSR), a G-protein receptor, is well recognized for its role in the regulation of adipocyte proliferation, in modulating adipose tissue dysfunction, and as a potential target for therapeutic intervention. In the present study, we investigate the anti-inflammatory effect of γ-glutamylvaline (γ-EV) on mouse adipocytes and explore the role of γ-EV-activated CaSR in the regulation of cellular homeostasis using the mouse 3T3-L1 cell line in vitro model. Our results indicate that the 3T3-L1 adipocyte-like cells accumulated lipids and expressed CaSR after 2 days of differentiation and 7 days of maturation period. The pretreatment with γ-EV (10 µM) suppressed the production of TNF-α-induced pro-inflammatory cytokines, i.e., IL-6 (23.92 ± 5.45 ng/mL, p < 0.05)) and MCP-1 (101.17 ± 39.93 ng/mL, p < 0.05), while enhancing the expression of PPARγ (1.249 ± 0.109, p < 0.001) and adiponectin (7.37 ± 0.59 ng/mL, p < 0.05). Elevated expression of Wnt5a was detected in γ-EV-treated cells (115.90 ± 45.50, p < 0.001), suggesting the involvement of the Wnt/ß-catenin pathway. Also, phosphorylation of ß-catenin was shown to be significantly inhibited (0.442 ± 0.034) by TNF-α but restored when cells were pretreated with γ-EV (0.765 ± 0.048, p < 0.05). These findings suggest that γ-EV-induced CaSR activation not only prevents TNF-α-induced inflammation in adipocytes but also modulates the cross-talk between Wnt and PPARγ pathways. Concentrations of serine phosphorylated IRS-1 were shown to be lower in γ-EV-treated cells, indicating γ-EV may also prevent inflammation in the context of insulin resistance. Thus, γ-EV-activated CaSR plays a significant role in the cross-talk between adipocyte inflammatory and metabolic pathways through the regulation of extracellular sensing.

Recent Advances in the Understanding of the Health Benefits and Molecular Mechanisms Associated with Green Tea Polyphenols.

Tea, leaf, or bud from the plant Camellia sinensis, make up some of the beverages popularly consumed in different parts of the world as green tea, oolong tea, or black tea. More particularly, as a nonfermented tea, green tea has gained more renown because of the significant health benefits assigned to its rich content in polyphenols. As a main constituent, green tea polyphenols were documented for their antioxidant, anti-inflammation, anticancer, anticardiovascular, antimicrobial, antihyperglycemic, and antiobesity properties. Recent reports demonstrate that green tea may exert a positive effect on the reduction of medical chronic conditions such as cardiovascular disease, cancer, Alzheimer's disease, Parkinson's disease, and diabetes. The health benefits of green teas, in particular EGCG, are widely investigated, and these effects are known to be primarily associated with the structure and compositions of its polyphenols. This Review focuses on the diverse constituents of green tea polyphenols and their molecular mechanisms from the perspective of their potential therapeutic function. Recent advances of green tea polyphenols on their bioavailability, bioaccessibility, and microbiota were also summarized in this article. Dietary supplementation with green tea represents an attractive alternative toward promoting human health.

A bioinformatics study on characteristics, metabolic pathways, and cellular functions of the identified S-nitrosylated proteins in postmortem pork muscle.

This study aimed to determine the characteristics, metabolic pathways and cellular functions of S-nitrosylated proteins from pork postmortem muscle using bioinformatics analysis. The results showed that S-nitrosylated proteins had a broad range of molecular weight and pI value and were mainly located in the functional region of secondary structure. The motif revealed the lysine (K) positioned at -5, -7, +1 and +5 through the S-nitrosocysteine while "C-X-X-C" was identified as the motif for non-S-nitrosylation-modified cysteine. The proteins were widely localized in cell compartments and mostly belonged to enzymes participating in the metabolic process. Glycolysis was the most significant pathways of S-nitrosylated proteins in postmortem muscle. The cell death of muscle cells was predicted to be inhibited by S-nitrosylation with the potential influence on the apoptosis. Those identified pathways and cellular functions of S-nitrosylation are proposed to have a profound influence on meat quality and should be highly regarded.

The proteomics homology of antioxidant peptides extracted from dry-cured Xuanwei and Jinhua ham.

The objective of this study was to investigate the antioxidant activity and the homology of peptides extracted from dry-cured Xuanwei and Jinhua ham. The antioxidant activity of crude peptides was assessed by ORAC and ABTS assays and the scavenging effects on DPPH and O2- free radicals. LC-ESI-Q-TOF-MS/MS was used to analyze the peptide composition. Based on the identified peptides, homologous proteins were matched by the Peaks software. Overall, 243 and 213 peptides were identified with their parent proteins in Xuanwei and Jinhua dry-cured hams. Based on the ORAC and TEAC values, XHP showed higher antioxidant ability than JHP (P < 0.05). After further purification by G-15 chromatography, the results indicate that the oligopeptides with less than 1000 Da had greater antioxidant activity than the other two fractions. Homology-based proteomics showed that the majority of peptides originated from myosin which accounted for 26% in XHP and 32% in JHP respectively.

Identification of S-nitrosylated proteins in postmortem pork muscle using modified biotin switch method coupled with isobaric tags.

The objective of this study was to identify the S-nitrosylated proteins in aging samples of pork longissimus thoracis muscle (aged 0 and 3 d) and to study the effects of exogenous S-nitrosoglutathione (GSNO, concentration at 10 and 100 µM) treatments of aged 0 d sample. After validating modified biotin switch method, the samples were labeled with tandem mass tags (TMT126-129) for the LC-MS/MS analysis. A total of 366 peptides were identified to be S-nitrosylated corresponding to 339 proteins. Comparison of total intensity and individual S-nitrosylated sites between aging samples revealed that S-nitrosylation did occur in pork muscle during postmortem aging through possible pathways of denitrosylation and transnitrosylation. GSNO treatment groups showed a considerable number of potential cysteines could be modified with high thiol-reactivity. It was deduced that S-nitrosylation could be involved in the postmortem metabolic process possibly through the regulation of activity or function of glycolytic enzymes, calcium release, heat shock proteins, antioxidant enzymes and myofibrillar proteins.

Effect of Flavourzyme on proteolysis, antioxidant activity and sensory qualities of Cantonese bacon.

The aim of this study was to investigate the influence of Flavourzyme on protein degradation, antioxidant activity and sensory qualities of Cantonese bacon made at 60°C for 28h. Flavourzyme was used at doses of 8, 16, 24, 32 and 40LAUP/kg of raw meat. Results indicated that Flavourzyme addition in Cantonese bacon promoted proteolysis, which was reflected by the increase of various free amino acids and SDS-PAGE-detected changes in the myogen and myofibrillar proteins. Sensory qualities were improved while TPA (texture profile analysis) was decreased significantly (P<0.05). The best sensory scores were obtained at between 24 and 32LAPU/kg. Besides, Flavourzyme addition improved antioxidant activity, and decreased water activity and protein carbonyl content of Cantonese bacon. This study indicated that the addition of Flavourzyme improved eating attributes and storage stability of Cantonese bacon.

A Review of Antioxidant Peptides Derived from Meat Muscle and By-Products.

Antioxidant peptides are gradually being accepted as food ingredients, supplemented in functional food and nutraceuticals, to positively regulate oxidative stress in the human body against lipid and protein oxidation. Meat muscle and meat by-products are rich sources of proteins and can be regarded as good materials for the production of bioactive peptides by use of enzymatic hydrolysis or direct solvent extraction. In recent years, there has been a growing number of studies conducted to characterize antioxidant peptides or hydrolysates derived from meat muscle and by-products as well as processed meat products, including dry-cured hams. Antioxidant peptides obtained from animal sources could exert not only nutritional value but also bioavailability to benefit human health. This paper reviews the antioxidant peptides or protein hydrolysates identified in muscle protein and by-products. We focus on the procedure for the generation of peptides with antioxidant capacity including the acquisition of crude peptides, the assessment of antioxidant activity, and the purification and identification of the active fraction. It remains critical to perform validation experiments with a cell model, animal model or clinical trial to eliminate safety concerns before final application in the food system. In addition, some of the common characteristics on structure-activity relationship are also reviewed based on the identified antioxidant peptides.

Identification of Rosmarinic Acid-Adducted Sites in Meat Proteins in a Gel Model under Oxidative Stress by Triple TOF MS/MS.

Triple TOF MS/MS was used to identify adducts between rosmarinic acid (RosA)-derived quinones and meat proteins in a gel model under oxidative stress. Seventy-five RosA-modified peptides responded to 67 proteins with adduction of RosA. RosA conjugated with different amino acids in proteins, and His, Arg, and Lys adducts with RosA were identified for the first time in meat. A total of 8 peptides containing Cys, 14 peptides containing His, 48 peptides containing Arg, 64 peptides containing Lys, and 5 peptides containing N-termini that which participated in adduction reaction with RosA were identified, respectively. Seventy-seven adduction sites were subdivided into all adducted proteins including 2 N-terminal adduction sites, 3 Cys adduction sites, 4 His adduction sites, 29 Arg adduction sites, and 39 Lys adduction sites. Site occupancy analyses showed that approximately 80.597% of the proteins carried a single RosA-modified site, 14.925% retained two sites, 1.492% contained three sites, and the rest 2.985% had four or more sites. Large-scale triple TOF MS/MS mapping of RosA-adducted sites reveals the adduction regulations of quinone and different amino acids as well as the adduction ratios, which clarify phenol-protein adductions and pave the way for industrial meat processing and preservation.