Protein extracted from Moringa oleifera Lam. Leaves: Bio-evaluation and characterization as suitable plant-based meat-protein alternative.

This study aimed to isolate and characterize moringa leaf protein (MLP) via HPLC and evaluate its consumption's effects through rat model. Four groups of Albino Wistar rats (n = 25 each) along with a control group (n = 25) were acclimatized. The isolated MLP was added to the basal diet (casein; control) in various percentages (25, 50, 75, 100%) for a 21-day experimental period. On three intervals (1st, 11th, 21st days), blood samples were collected and subjected for hematological and biochemical examination (Renal Function Test (RFT), Liver Function Test (LFT)). MLP contained a variety of essential and non-essential amino acids in substantial amounts. The Protein Efficiency Ratio (PER) of 50% MLP-treated group was the highest (1.72) among MLP treatments. Increases in feed intake and weight were observed in treated rats compared to the control. The hematological profile of the rats revealed increases in Hemoglobin (Hb) (7.9-14.0%), White Blood Cell (WBC) (35.9-51.5%), Red Blood Cell (RBC) (17.1-22.2%), Hematocrit (HCT) (13.1-22.9%), and platelets levels (36.5-40.6%) from day 1. Protein isolates decreased liver parameters but resulted in non-significant changes in liver and kidney functions in rats. Further investigation is needed to determine the safe daily intake of MLP.

Line-scan Raman scattering image and multivariate analysis for rapid and noninvasive detection of restructured beef.

The mean spectral (MS) features were extracted from Raman scattering images (RSI) of beef samples over the region of interest covering the spectral range of 789-1710cm-1 and the spatial offset range of 0-5 mm (for two sides of the incident laser). The RSI monitored the main change in the protein, amide bands, lipids, and amino acid residues. The classification model performance based on MS features compared the conventional Raman spectral features and confirmed the usefulness of RSI. Finally, the results showed that RSI technology is a reliable tool for rapid and noninvasive detection of restructured beef.

Multispecies Identification of Oilseed- and Meat-Specific Proteins and Heat-Stable Peptide Markers in Food Products.

Consumer demand for both plant products and meat products enriched with plant raw materials is constantly increasing. Therefore, new versatile and reliable methods are needed to find and combat fraudulent practices in processed foods. The objective of this study was to identify oilseed species-specific peptide markers and meat-specific markers that were resistant to processing, for multispecies authentication of different meat and vegan food products using the proteomic LC-MS/MS method. To assess the limit of detection (LOD) for hemp proteins, cooked meatballs consisting of three meat species and hemp cake at a final concentration of up to 7.4% were examined. Hemp addition at a low concentration of below 1% was detected. The LOD for edestin subunits and albumin was 0.9% (w/w), whereas for 7S vicilin-like protein it was 4.2% (w/w). Specific heat-stable peptides unique to hemp seeds, flaxseed, nigella, pumpkin, sesame, and sunflower seeds, as well as guinea fowl, rabbit, pork, and chicken meat, were detected in different meat and vegan foods. Most of the oilseed-specific peptides were identified as processing-resistant markers belonging to 11S globulin subunits, namely conlinin, edestin, helianthinin, pumpkin vicilin-like or late embryogenesis proteins, and sesame legumin-like as well as 2S albumins and oleosin isoforms or selected enzymic proteins.

Current and future prospects for the use of pulsed electric field in the meat industry.

Pulsed electric field (PEF) is a novel non-thermal technology that has recently attracted the attention of meat scientists and technologists due to its ability to modify membrane structure and enhance mass transfer. Several studies have confirmed the potential of pulsed electric field for improving meat tenderness in both pre-rigor and post-rigor muscles during aging. However, there is a high degree of variability between studies and the underlying mechanisms are not clearly understood. While some studies have suggested physical disruption as the main cause of PEF induced tenderness, enzymatic nature of the tenderization seems to be the most plausible mechanism. Several studies have suggested the potential of PEF to mediate the tenderization process due to its membrane altering properties causing early release of calcium ions and early activation of the calpain proteases. However, experimental research is yet to confirm this postulation. Recent studies have also reported increased post-mortem proteolysis in PEF treated muscles during aging. PEF has also been reported to accelerate curing, enhance drying and reduce the numbers of both pathogens and spoilage organisms in meat, although that demands intense processing conditions. While tenderization, meat safety and accelerated curing appears to be the areas where PEF could provide attractive options in meat processing, further research is required before the application of PEF becomes a commercial reality in the meat industry. It needs to deal with carcasses which vary biochemically and in composition (muscle, fat, and bones). This review critically evaluates the published reports on the topic with the aim of reaching a clear understanding of the possible applications of PEF in the meat sector in addition to providing some insight on critical issues that need to be addressed for the technology to be a practical option for the meat industry.

Mass Spectrometry-Based Immunoassay for the Quantification of Banned Ruminant Processed Animal Proteins in Vegetal Feeds.

The ban of processed animal proteins (PAPs) in feed for farmed animals introduced in 2001 was one of the main EU measures to control the bovine spongiform encephalopathy (BSE) crisis. Currently, microscopy and polymerase chain reaction (PCR) are the official methods for the detection of illegal PAPs in feed. However, the progressive release of the feed ban, recently with the legalization of nonruminant PAPs for the use in aquaculture, requires the development of alternative methods to determine the species origin and the source (legal or not). Additionally, discussions about the need for quantitative tests came up, particularly if the zero-tolerance-concept is replaced by introducing PAP thresholds. To address this issue, we developed and partially validated a multiplex mass spectrometry-based immunoassay to quantify ruminant specific peptides in vegetal cattle feed. The workflow comprises a new sample preparation procedure based on a tryptic digestion of PAPs in suspension, a subsequent immunoaffinity enrichment of the released peptides, and a LC-MS/MS-based analysis for peptide quantification using isotope labeled standard peptides. For the very first time, a mass spectrometry-based method is capable of detecting and quantifying illegal PAPs in animal feed over a concentration range of 4 orders of magnitude with a detection limit in the range of 0.1% to 1% (w/w).

ε-Polylysine-mediated enhancement of the structural stability and gelling properties of myofibrillar protein under oxidative stress.

The effects of ε-polylysine (ε-PL) at different concentrations (0.005 %, 0.010 %, 0.020 %, and 0.030 %) on the structure and gelling behavior of pork myofibrillar protein (MP) under oxidative stress were explored. The incorporation of ε-PL significantly restrained oxidation-induced sulfhydryl and solubility losses (up to 9.72 % and 41.9 %, respectively) as well as protein crosslinking and aggregation. Compared with the oxidized control, ε-PL at low concentrations (0.005 % - 0.020 %) promoted further unfolding and destabilization of MP, while 0.030 % ε-PL led to refolding of MP and enhanced its thermal stability. The ε-PL-induced physicochemical changes favored the formation of a finer and more homogeneous three-dimensional network structure, therefore obviously enhancing the strength and water-holding capacity (WHC) of thermally induced oxidized MP gels, with the ε-PL at 0.020 % showed the greatest enhancement. This work revealed for the first time that ε-PL can significantly ameliorate the oxidation stability and gel-forming ability of meat proteins.

The influence of age and gender on the quality of raw and roasted wild boars (Sus scrofa) meat.

The aim of this study was to determine the influence of age and gender on the chemical composition, colour, texture and sensory attributes of wild boars meat (Sus scrofa, n = 48). The animals were analysed in 6 groups (n = 8), differed in age (piglets below 1 year, yearlings 1-2 years, and adults over 2 years) and gender (male and female). Protein content in raw meat was affected by age, whereas in roasted meat by animals gender. Colour parameters, excluding lightness, were affected by animals age and values of a*, b* and C increased along the animal age. Roasted meat from piglets showed the lowest hardness and chewiness in TPA test, however shear force was similar for piglets and yearling, yet lower than adults. Age affected sensory quality of roasted meat and meat from yearlings was scored higher than from adults. The results of the study clearly indicate the predominant influence of wild boar age over gender on the meat quality.

Effects of soy protein isolate on gel properties and water holding capacity of low-salt pork myofibrillar protein under high pressure processing.

This paper studies the effects of soy protein isolate (SPI; 0, 2% and 4%; Weight/Weight) on texture, rheological property, sulfhydryl groups, and the water distribution state of low-salt (1% NaCl) pork myofibrillar protein systems under high pressure processing (HPP, 200 MPa, 10 min). The L⁎ value, cooking yield, hardness, total and reactive sulfhydryl, surface hydrophobicity, and the G' value at 80 °C of pork myofibrillar protein increased significantly (P < 0.05) when SPI was added; however, the springiness, cohesiveness, and chewiness of gels with 4% SPI were lower than of gels with 2% SPI. The rheological findings indicated that the thermal stability of the myofibrillar protein increased when SPI was added. The initial relaxation time of T2b, T21, and T22 decreased when SPI increased; meanwhile, the peak ratio of P21 increased significantly (P < 0.05), implying that water had lower mobility. Overall, the 2% SPI could enhance gel characteristics and water-holding capacity of pork myofibrillar protein under 200 MPa.

Optimal raw material formulation to produce pressed pork ham at the least cost.

This study optimized material use for making pressed pork hams (PPHs) using the least cost formulation program. Based on protein (P) content, different fat (2.5, 3.0, and 4.0P) and moisture (4.0P + 10, 4.0P + 15, 4.0P + 20, and 4.0P + 25) ratios were applied to make PPHs. Total expressible fluid, cooking loss, and purge loss were highest in PPHs formulated with 4.0P + 25 water (P < 0.05). With increasing fat ratio, lightness increased, but redness decreased (P < 0.05). Lipid oxidation was not affected by moisture ratio (P > 0.05). Hardness, gumminess, and adhesiveness decreased with increased fat and moisture (P < 0.05). PPH cost was reduced by fat and moisture. These results suggest that the formulations with 4.0P + 15 moisture, regardless of fat ratio, are recommended for PPHs due to their moderate cost and quality characteristics. In addition, greater cost savings can be expected by further subdividing raw material criteria for industrial applications.

Proteolysis and protein oxidation throughout the smoked dry-cured ham process.

During the five stages of smoked dry-cured ham processing, proteolysis and protein oxidation were simultaneously detected in the Biceps femoris (BF) and Semimembranosus (SM) muscles. Proteolysis was more advanced in BF than in SM throughout the process of production. The total FAA increased significantly (p < 0.05) throughout the processing, resulting in higher total FAA content in BF than in SM muscle. SDS-PAGE revealed progressive degradation of sarcoplasmic proteins of investigated muscles, with the pronounced changes for the 69.9-41.7 kDa region. SDS-PAGE of BF showed more intense degradation of myofibrillar proteins due to greater proteolysis in BF. Electrophoresis of myofibrillar proteins evidenced the marked degradation of 130 kDa, 96.7 kDa and 27-20.7 kDa bands in both muscles. A similar trend was observed for protein oxidation in BF and SM, with the final values of 26.36 and 23.7 nmol carbonyls/mg proteins, respectively. The Pearson correlation revealed a strong relationship between protein oxidation and proteolysis.

Regulation of protein oxidation in Cantonese sausages by rutin, quercetin and caffeic acid.

The effects of rutin, quercetin, and caffeic acid on protein oxidation in Cantonese sausage during 60 days of storage at room temperature (25 ± 1 °C) were investigated. The results showed that the three phenolic compounds played different roles in inhibiting the oxidation of sarcoplasmic and myofibrillar protein. All of them inhibited sarcoplasmic protein oxidation by retarding carbonylation, the conversion of SH to S-S groups, and the formation of dimeric tyrosine and Schiff bases, of which rutin is the most effective. For myofibrillar protein, all of them suppressed the conversion of SH to S-S groups, only caffeic acid inhibited the accumulation of Schiff bases instead of carbonyls while both quercetin and caffeic acid inhibited the formation of dimeric tyrosine. In addition, quercetin had an inverse dosage effect on the oxidation regulation of MP, 0.16 g/kg quercetin had better inhibit effect on protein oxidation than 0.32 g/kg quercetin.

Effects of Casein, Chicken, and Pork Proteins on the Regulation of Body Fat and Blood Inflammatory Factors and Metabolite Patterns Are Largely Dependent on the Protein Level and Less Attributable to the Protein Source.

The impact of meat protein on metabolic regulation is still disputed and may be influenced by protein level. This study aimed to explore the effects of casein, pork, and chicken proteins at different protein levels (40% E vs 20% E) on body weight regulation, body fat accumulation, serum hormone levels, and inflammatory factors/metabolites in rats maintained on high-fat (45% E fat) diets for 84 d. Increased protein levels resulted in a significant reduction in body fat mass and an increase in the serum levels of the anti-inflammatory cytokine IL-10, independent of protein source. Analysis of blood via untargeted metabolomics analysis identified eight, four, and four metabolites significantly altered by protein level, protein source, and a protein level-source interaction, respectively. Together, the effects of casein, chicken, and pork protein on the regulation of body fat accumulation and blood metabolite profile are largely dependent on protein level and less attributable to the protein source.

TMT-based quantitative proteomic analysis of porcine muscle associated with postmortem meat quality.

To explore the molecular mechanisms of meat quality, four high-quality (HQ) samples and four low-quality (LQ) samples from longissimus dorsi muscles were chosen, and tandem mass tag (TMT) labeling combined with mass spectrometry (MS) were performed to find associations between meat quality and proteome profiles. The LQ meats had lower pH, lighter color, and higher drip loss compared to the HQ meats. About 140 differentially expressed proteins were identified. Functional analysis results of differentially expressed proteins showed that decreased release of Ca2+, lower contents of type II fibers, lower contents of glycogen, and decreased glycogenolysis in HQ meats indicated a lower degree of glycolysis in HQ as compared to LQ meats. Meanwhile, some differentially expressed proteins suggested that the levels of oxidative stress and apoptosis were lower in HQ meats than in LQ meats. This study reveals physiological changes between HQ and LQ meats according to the proteome profiles.

Meat Protein in High-Fat Diet Induces Adipogensis and Dyslipidemia by Altering Gut Microbiota and Endocannabinoid Dysregulation in the Adipose Tissue of Mice.

Endocannabinoids modulate insulin and adipokine expression in adipocytes through cannabinoid receptors and their levels are elevated during hyperglycemia and obesity, but little is known about how diets affect them. We assessed the effects of dietary casein, chicken, beef, and pork proteins in a high-fat diet mode, on endocannabinoids, adipogenesis, and biomarkers associated with dyslipemdia. A high-fat beef or chicken diet upregulated cannabinoid 1 receptor, N-acyl phosphatidylethanolamine-selective phospholipase-D and diacylglycerol lipase α in adipose tissue and reduced the immunoreactivity of mitochondrial uncoupling protein 1 in brown adipose tissue. In addition, the high-fat diets with beef and chicken protein had a significant impact on adipocyte differentiation and mitochondrial biogenesis in obese mice. A 16S rRNA gene sequencing indicated that high-fat diets, regardless of the protein source, significantly enhanced the ratio of Firmicutes to Bacteroidetes in colon. Meat proteins in a high-fat diet significantly decreased the relative abundances of Akkermansia and Bifidobacteria but enhanced the lipopolysaccharides level in the serum, which promoted the adipogenesis process by causing dysregulation in the endocannabinoid receptors. Consumption of meat protein with high-fat-induced adiposity, visceral obesity, and dyslipidemia reduced the thermogenesis and had a distinctive effect on the mitochondrial biogenesis compared with casein protein.

Optimization of the protein extraction method of goat meat using factorial design and response surface methodology.

Protein extraction from goat meat was carried out based on the combination of response surface methodology and factorial design to optimize the variables: temperature (25-50 °C), extraction time (8-20 min), volume (3-10 mL) and extractor concentration (0.05-0.1 mol L-1). The proposed model did not present a lack of fit, explaining 96% of the total data variance (R2 = 0.96). The optimum extraction conditions were: 0.05 mol L-1 for extractor concentration, extraction time of 10 min, temperature of 44 °C and extractor volume of 3.5 mL. The protein content (19.3 g/100 g) obtained by the optimized method was higher than some results reported in the literature. HPLC-SEC-DAD analysis revealed that the extraction conditions used did not significantly modify the protein structure. The proposed method proves to be simple, fast, robust, cheap and adequate for native protein extraction, being a potential approach for proteomic research focusing in goat meat.

Sparse NIR optimization method (SNIRO) to quantify analyte composition with visible (VIS)/near infrared (NIR) spectroscopy (350 nm-2500 nm).

Visual-Near-Infra-Red (VIS/NIR) spectroscopy has led the revolution in high-throughput phenotyping methods used to determine chemical and structural elements of organic materials. In the current state of the art, spectrophotometers used for imaging techniques are either very expensive or too large to be used as a field-operable device. In this study we developed a Sparse NIR Optimization method (SNIRO) that selects a pre-determined number of wavelengths that enable quantification of analytes in a given sample using linear regression. We compared the computed complexity time and the accuracy of SNIRO to Marten's test, to forward selection test and to LASSO all applied to the determination of protein content in corn flour and meat and octane number in diesel using publicly available datasets. In addition, for the first time, we determined the glucose content in the green seaweed Ulva sp., an important feedstock for marine biorefinery. The SNIRO approach can be used as a first step in designing a spectrophotometer that can scan a small number of specific spectral regions, thus decreasing, potentially, production costs and scanner size and enabling the development of field-operable devices for content analysis of complex organic materials.

Effects of oxidative modification on textural properties and gel structure of pork myofibrillar proteins.

Isolated myofibrillar protein (MP) was treated by the oxidation system of FeCl3 (0.01 mM) at four different H2O2 concentrations (0, 1, 10, 20 mM). The oxidation degree was determined by measuring the carbonyl and total sulphydryl values. The structure and physicochemical properties of MP gels were investigated by water holding capacity (WHC) evaluation, sodium dodecyl sulfate-polyacryl amide gel electrophoresis (SDS-PAGE), texture profile analysis (TPA), Raman spectroscopy, and NMR transverse relaxation (T2). The results of carbonyls and total sulphydryls indicated that oxidation degree of MP increased with increasing H2O2 concentration. TPA showed that moderate oxidation (10 mM H2O2) could improve the hardness, springiness, gumminess and cohesiveness of MP gels, but not contribute to the maintenance of their WHC, probably due to severe depolymerization of MPs, unfolding of α-helix, exposure of the hydrophobic groups and the migration of protein-associated water (T2b) and intra-myofibrillar water (T21) to the longer relaxation time.

El Gueddid, a traditional Algerian dried salted meat: Physicochemical, microbiological characteristics and proteolysis intensity during its manufacturing process and ripening.

El Gueddid is a traditional salted and dried meat with high popularity in Algeria. It is used as an ingredient in various dishes. In this study, different samples of El Gueddid were analyzed at different processing times to follow up their microbiological and physicochemical properties. Changes in the protein profile were also demonstrated by electrophoretic study of myofibrillar proteins. Microbiological determinations included the total viable count, coliforms, Staphylococci, lactic acid bacteria, yeasts, and molds, whereas physicochemical properties were characterized by pH, moisture, salt content and water activity. The results showed that microbial profiles were elevated for all the studied micro-organisms. Staphylococci and lactic acid bacteria were the most abundant micro-organisms in the product. Total coliforms were found in low numbers in fresh meat, being eliminated at the post salting stage of process. The physicochemical characteristics showed that the moisture content decreased in the product during the drying period. The pH also decreased during the drying period, then remained almost unchanged during the rest of the ripening period. Moreover, El Gueddid showed low water activity and high salt content. One of the most important changes in the profile of myofibrillar proteins was a reduction in the myosin heavy chain content.

Specific Microbiota Dynamically Regulate the Bidirectional Gut-Brain Axis Communications in Mice Fed Meat Protein Diets.

The purpose of this study was to characterize the dynamic changes of different protein diets to gut microbiota and explore the influence on communications between the gut and the brain. C57BL/6J mice were fed casein, soy protein, and four kinds of processed meat proteins at a normal dose of 20% for 8 months. Bacteroidales S24-7 abundance increased from 4 to 8 months, whereas the abundances of six genera including Akkermansia decreased remarkably. Lachnospiraceae Unclassified abundance in the emulsion-type sausage protein and stewed pork protein groups showed an opposite change from 4 to 8 months. Twenty-eight and 48 specific operational taxonomy units in cecum and colon respectively were involved in regulating serotonin, peptide YY, leptin, and insulin levels. Specific microbiota was involved, directly or indirectly through signaling molecules, in the regulation of body metabolism, which may affect the communications between the gut and brain and cause different growth performances.

Combined effect of CaCl2 and high pressure processing on the solubility of chicken breast myofibrillar proteins under sodium-reduced conditions.

The combined effect of calcium chloride (CaCl2) (20-100 mM) and high pressure processing (HPP, 200 MPa) on the solubility of myofibrillar proteins (MP) was investigated under sodium-reduced conditions (0.3 M sodium chloride). The results revealed that HPP combined with low concentrations (<40 mM) of CaCl2 synergistically increased the solubility of MP, but an antagonistic effect occurred when a high concentration (100 mM) of CaCl2 was present. This synergistic effect could be attributed to a mildly destabilized conformation of myosin, an increased surface hydrophobicity and the decreased total sulfhydryl group content of MP. However, CaCl2 at 100 mM destabilized myosin to a larger extent and induced non-disulfide covalent cross-linking of S-1 subfragment in pressurized MP, thus attenuating the solubilizing effect of HPP on the myosin heavy chain, resulting in the antagonistic effect. Therefore, HPP in combination with low-level CaCl2 (<40 mM) may improve the functional properties of sodium-reduced meat products.