High post-mortem temperature combined with rapid glycolysis induces phosphorylase denaturation and produces pale and exudative characteristics in broiler Pectoralis major muscles

This study investigated the effect of early post-mortem temperature on broiler protein characteristics and meat quality. Muscles were kept at different temperatures (0, 20 and 40 °C) until 4h post-mortem and then stored at 4 °C. Rapid degradation of ATP and glycogen, thus inducing a high rate of lactate formation and pH drop, were found in the 40 °C group during incubation. When extracting proteins, a lower protein content of the sarcoplasmic fraction and a higher protein content of the myofibrillar fraction were found in the 40 °C group at 24h post-mortem; SDS-PAGE and western-blotting results revealed that phosphorylase was associated with the myofibrillar fraction. Furthermore, the 40 °C group had paler surfaces, higher drip loss and lower processing properties. These data suggest that elevated temperature during early post-mortem period, resulting in rapid glycolysis, induced phosphorylase denaturation and association with myofibrillar proteins thus generating pale and exudative characteristics.     

Effect of pre-rigor temperature incubation on sarcoplasmic protein solubility,calpain activity and meat properties in porcine muscle

The aim of the study was to investigate the effect of pre-rigor temperature incubation on sarcoplasmic protein characteristics in relation to meat properties within porcine muscle. Porcine Longissimus dorsi muscles were incubated at temperatures of 0, 10, 20, 30 and 40 °C to 6 h post mortem. Incubation at 40 °C induced a significant decrease of sarcoplasmic protein solubility and an increase in proteins in the myofibrillar fraction. The protein relocation was followed till 72 h post mortem but had largely been completed by the end of the temperature incubation at 6 h. SDS-PAGE and Western blot analyses suggested that phosphorylase and creatine kinase precipitated onto the myofilaments during incubation at 40 °C. Drip loss increased following incubation at 40 °C, indicating that the precipitation of phosphorylase and creatine kinase may be a factor of reduced water-holding capacity at the combination of high temperature and low pH. Incubation at 40 °C resulted in substantially lower shear force in parallel with loss of extractable activity of μ- and m-calpain, suggesting a rapid activation of both enzymes at high temperatures and low pH early post mortem.     

Temperature induced denaturation of myosin: Evidence of structural alterations of myosin subfragment-1

Denaturation of myofibrillar proteins in porcine longissimus thoracis et lumborum muscle was investigated after pre-rigor temperature incubation at 20, 30 and 40 °C. At 24 h myofibrils were isolated and myosin was further cleaved by chymotrypsin. High temperature pre-rigor induced release of myosin S1 (subfragment-1), less (P < 0.05) Ca²⁺-ATPase activity and structural alterations of the region of the myosin molecule that harbors S1. Surface hydrophobicity of myofibrils from the 40 °C group increased (P < 0.001), suggesting a temperature-induced structural rearrangement exposing hydrophobic groups on the surface of myofibrils which in turn may explain the reduced water-holding of PSE meat.     

A simple,reliable and reproductive method to obtain experimental pale,soft and exudative (PSE) pork

The inferior quality and economic risk of pale, soft and exudative (PSE) pork warrant continuing research. However, such research efforts are often hindered by the challenge to obtain reliable PSE muscle samples with similar quality characteristics. The objective of this study was to establish a reliable and convenient method to produce PSE-like pork. A PSE condition was induced by incubation of 30-min postmortem Longissimus muscle at 35 °C for 7 h followed by chilling to 4 °C. Compared to normal red, firm and non-exudative (RFN) pork (kept at 4 °C), PSE muscle had consistently lower pH_2h (5.46 vs. 5.74) and pH_4h (5.35 vs. 5.52), higher L* (lightness) value (56.5 vs. 51.0), and reduced protein solubility and thermal stability (enthalpy and temperature) than RFN muscle (P < 0.05). The highly reproducible results indicate that incubation of muscle immediately postmortem at 35 °C offers a simple and consistent method to produce PSE pork.     

Influence of early pH decline on calpain activity in porcine muscle

This study investigated the influence of post-mortem pH decline on calpain activity and myofibrillar degradation. From 80 pigs, 30 Longissimus dorsi (LD) muscles were selected on the basis of pH values at 3 h post-mortem and classified into groups of 10 as fast, intermediate and slow pH decline. The rate of pH decline early post-mortem differed between the three groups, but the ultimate pH values were similar at 24 h. Calpain activity and autolysis from 1 to 72 h post-mortem were determined using casein zymography and studied in relation to myofibrillar fragmentation.     

Biochemical and physicochemical properties of thermally treated natural actomyosin extracted from normal and PSE pork Longissimus muscle

Biochemical and physicochemical properties of thermally treated natural actomyosin (NAM) from normal and pale, soft and exudative (PSE) pork were studied. About 37% and 25% of available sulphydryl groups formed disulphide bonds or other permanent chemical bonds at 70 °C in NAM from normal and PSE pork, respectively. Surface hydrophobicities of NAM from normal and PSE pork at 70 °C were 3.6 and 2.4 times greater than that at 40 °C. About 90% of the α-helical structure of NAM was lost by heating to 70 °C. The temperature at maximum α-helical content decline of NAM was in accordance with the peak 3 thermal transition obtained by differential scanning calorimetry and the lowest storage modulus (G′) during thermal rheology. NAM from normal pork underwent aggregation with a higher extent of hydrophobic interaction and disulphide bonds, higher temperatures at maximum velocity for conformational change and unfolding than that from PSE pork. As a consequence, NAM from normal pork had superior rheological properties.     

Influence of muscle type on rheological properties of porcine myofibrillar protein during heat-induced gelation

The gelation characteristics of myofibrillar proteins are indicative of meat product texture. Defining the performance of myofibrillar proteins during gelation is beneficial in maintaining quality and developing processed meat products and processes. This study investigates the impact of pH on viscoelastic properties of porcine myofibrillar proteins prepared from different muscles (semimembranosus (SM), longissimus dorsi (LD) and psoas major (PM)) during heat-induced gelation. Dynamic rheological properties were measured while heating at 1 °C/min from 20 to 85 °C, followed by a holding phase at 85 °C for 3 min and a cooling phase from 85 to 5 °C at a rate of 5 °C/min. Storage modulus (G′, the elastic response of the gelling material) increased as gel formation occurred, but decreased after reaching the temperature of myosin denaturation (52 °C) until approximately 60 °C when the gel strength increased again. This resulted in a peak and depression in the thermogram. Following 60 °C, the treatments maintained observed trends in gel strength, showing SM myofibrils produced the strongest gels. Myofibrillar protein from SM and PM formed stronger gels at pH 6.0 than at pH 6.5. Differences may be attributed to subtle variations in their protein profile related to muscle type or postmortem metabolism. Significant correlations were determined between G′ at 57, 72, 85 and 5 °C, indicating that changes affecting gel strength took effect prior to 57 °C. Muscle type was found to influence water-holding capacity to a greater degree than pH.     

Influence of salt and pH on the solubility and structural characteristics of transglutaminase-treated wheat gluten hydrolysate

Hydrolyzed wheat gluten (GH, 77–85% protein) was prepared by limited chymotrypsin digestion at 37 °C for 4 h (degree of hydrolysis = 6.4%) and 15 h (degree of hydrolysis = 10.3%). Microbial transglutaminase (MTGase) treatment (55 °C for 1 h, or 5 °C for 18 h) effect on the solubility and structural characteristics of GH was examined under selected food processing conditions (pH 4.0–7.0, 0–0.6 M NaCl). The MTGase treatment increased solubility of GH by 3–29-fold (P < 0.05) within pH 4.0–7.0. Addition of 0.6 M NaCl or changing the conditions of MTGase incubation did not significantly alter solubility characteristics of GH. The MTGase treatment decreased surface hydrophobicity, and increased carboxyl groups in GH, suggesting cross-linking and deamidation. Fluorescence and UV spectra attributed the improved GH solubility to MTGase-induced polar environment, and partial masking of some nonpolar aromatic amino acids possibly due to high-molecular-weight polypeptides formed.     

High pressure induced changes in beef muscle proteome: Correlation with quality parameters

The relationship between pressure induced changes on individual proteins and selected quality parameters in bovine longissimus thoracis et lumborum (LTL) muscle was studied. Pressures ranging from 200 to 600 MPa at 20 °C were used. High pressure processing (HPP) at pressures above 200 MPa induced strong modifications of protein solubility, meat colour and water holding capacity (WHC). The protein profiles of non-treated and pressure treated meat were observed using two dimensional electrophoresis. Proteins showing significant differences in abundance among treatments were identified by mass spectrometry. Pressure levels above 200 MPa strongly modified bovine LTL proteome with main effects being insolubilisation of sarcoplasmic proteins and solubilisation of myofibrillar proteins. Sarcoplasmic proteins were more susceptible to HPP effects than myofibrillar. Individual protein changes were significantly correlated with protein solubility, L*, b* and WHC, providing further insights into the mechanistic processes underlying HPP influence on quality and providing the basis for the future development of protein markers to assess the quality of processed meats.     

Protein extractability and thermal gel formability of myofibrils isolated from skeletal and cardiac muscles at different post-mortem periods

The effects of the post-mortem ageing period on the extractability of myofibrillar proteins from pork cardiac and rabbit skeletal muscles under various conditions of pH and ionic strength were studied with particular reference to the changes in the solubility of individual myofibrillar proteins and their denaturation characteristics. The ultimate influence of these changes on the heat-induced gel forming ability of myofibrils isolated from cardiac and skeletal muscles at different post-mortem stages was also investigated. Results showed that pork cardiac myofibrils always exhibited lower solubility than those from rabbit skeletal muscles under identical conditions of pH, ionic strength and temperature. SDS-PAGE profiles indicated several quantitative differences in the relative proportion of individual protein species present in cardiac and skeletal myofibrils. The solubility of various proteins present in myofibrils was also affected differently on heating in 0-1 and 0-6 _M NaCl solution at various pH values. Thermal denaturation of cardiac myofibrils occurred at about 10°C higher than that of skeletal myofibrils as revealed by differential scanning calorimetry. Cardiac myofibrils formed much weaker heat-induced gels than those produced by skeletal myofibrils under identical conditions of temperature, pH, ionic strength and protein content.     

Effect of porcine plasma protein and setting on gel properties of surimi produced from fish caught in Thailand

Effects of porcine plasma protein (PPP) and high temperature setting on gel properties of surimi from bigeye snapper, bigeye croaker, threadfin bream and barracuda were investigated. PPP was effective in increasing breaking force and deformation of kamaboko gels set at 40°C for 30 min and heated at 90°C for 20 min. The optimum levels of PPP were 0.5, 0.5, 1.5 and 1.5 g/100 g and the optimum setting times were 2, 1.5, 1.5 and 2 h for bigeye snapper, bigeye croaker, threadfin bream and barracuda surimi, respectively. However, the addition of PPP significantly decreased whiteness (P<0.05). An increase in gel-forming ability of surimi with PPP coincided with a decrease in solubility in mixture of SDS, urea and β-mercaptoethanol, indicating the formation of nondisulfide covalent bond induced by both endogenous and plasma transglutaminase. The results supported that PPP improve the gelation of surimi in combination with setting.     

Increasing extractability of protein for allergen detection after food processing

The food industry is concerned about the presence of hidden allergens or undesired protein residues in final products and has to take measures to comply with current food allergen-labelling regulations. It is known that thermal processing decreases protein solubility and hinders the accuracy of their quantitation. The objective of this study was to optimise the extraction step before quantitation. Almonds were roasted at several temperatures (230, 260 and 400 °C) for 10 min and ground to meals, defatted and extracted with water, PBS and PBST for 30 min at 25 °C, 40 °C, 60 °C and 70 °C, respectively. The extracts were then sonicated in a temperature-controlled water bath and aliquots were taken after 0, 1, 3, 5 and 10 min of sonication. Ultrasonic treatment improved protein extraction from almonds, especially those roasted at 260 and 400 °C. The temperature of the extraction buffer proved to be important, with higher yields at 60 and 70 °C. Use of a higher temperature, with sonication for 5–10 min, increased the amount of protein extracted with PBST (from 141% to 174% for almonds roasted at 260 °C and 287% to 333% for almonds roasted at 400 °C). The BCA assay was used to measure the amount of soluble almond protein. Improved extraction of almond proteins from processed samples is presented.     

Molecular Properties of Post-Mortem Muscle. 6. Effect of Temperature on Protein Solubility of Rabbit and Bovine Muscle

SUMMARY– Studies were conducted to investigate the effect of temperature on the actin-myosin interaction of rabbit and bovine muscle during rigor and post-rigor shortening. Muscle was stored at four different temperatures (2°, 16°, 25° and 37°), corresponding to three types of post-mortem muscle shortening: cold, minimal and high temperature. These three types of shortening are presumably related to different states of the actin-myosin interaction in post-mortem muscle. Post-mortem tenderization may be the result of either actin-myosin dissociation or F-actin depolymerization.
To detect the occurrence of either of these possible changes, two salt solutions, differing widely in their myofibrillar protein extracting abilities, were used to compare post-mortem myofibrillar protein solubility after different times of post-mortem storage and to provide information about the actin-myosin complex. Myofibrillar protein solubility of both rabbit and beef muscle in 0.5M KCl, 0.1M phosphate, pH 7.4, increased markedly with increasing post-mortem storage at temperatures up to 25deg;. Similar solubility changes were obtained with 1.1M Kl, 0.1M K phosphate, pH 7.4, but these changes were much smaller in magnitude. Solubility in both salt solutions, in general, decreased for muscle stored at 37°.
Although time and temperature of post-mortem storage caused appreciable alterations in protein solubility, these alterations could not be directly related to changes in tenderness or sarcomere length or to species differences in the effects of temperature on post-mortem shortening. Viscosity, analytical ultracentrifugation, and ATPase assays all indicated the absence of "normal" actomyosin in all myofibrillar protein extracts in this study. It was suggested that the 1.1 M KI extracts contained G-actomyosin, but the available evidence indicated the presence of only myosin in 3-hr, 0.5 M KCI extracts.     

Emulsifying and foaming properties of transglutaminase-treated wheat gluten hydrolysate as influenced by pH,temperature and salt

Hydrolyzed wheat gluten (GH, 77–85% protein) was prepared by limited hydrolysis with chymotrypsin at 37 °C for 4 h (degree of hydrolysis=6.4%) and 15 h (degree of hydrolysis=10.3%). The effect of microbial transglutaminase (MTGase) treatment (55 °C for 1 h, or 5 °C for 18 h) on the emulsifying and foaming properties of GH was evaluated under selected food processing conditions (pH 4.0 and 6.5, 0 and 0.6 M NaCl, and temperature 20 and 5 °C). At pH 4.0 and 0 M NaCl the MTGase treatment substantially increased foaming capacity (FC) of GH compared with their respective control GH samples, as a result of enhanced peptide adsorption to the air–water interface, but FC was similar for both control and MTGase-treated GH at pH 6.5. In contrast, foam drainage stability (FS) of MTGase-treated GH decreased at pH 4.0, but increased significantly (P<0.05) at pH 6.5 when compared with their respective control GH samples. The FC and FS were affected by 0.6 M NaCl in a pH-dependent manner. The MTGase treatments increased emulsion activity index up to 15-fold at pH 6.5, while emulsion stability index was influenced by emulsion temperature and ionic strength conditions. The MTGase-induced changes in functional properties of GH were attributed to pH-dependent solubility changes, the amphiphilic nature of gluten peptides and increased electrostatic repulsion resulting from deamidation.     

Effect of fermentation temperature on the microbial and physicochemical properties of silver carp sausages inoculated with Pediococcus pentosaceus

The effect of fermentation with Pediococcus pentosaceus at different temperatures ranging from 15 to 37 °C on the quality characteristics of silver carp sausages was investigated. Higher temperature stimulated the rapid growth of lactic acid bacteria, resulting in a rapid decline in pH, and consequently suppressed the growth of Pseudomonas, Micrococcaceae and Enterobacteriaceae. However, increasing fermentation temperature gave a progressive increase in total volatile basic nitrogen and biogenic amines in fermented silver carp sausages. Histamine was the main biogenic amine, exceeding 100 mg/kg after 48 h of fermentation at temperatures above 30 °C. Higher content of non-protein nitrogen and α-amino nitrogen correlated with the electrophoretic studies, which showed that proteolysis of high molecular weight myofibrillar and sarcoplasmic proteins was more prominent at higher fermentation temperatures. Products fermented at 23–30 °C showed greatest consumer preference and most favourable textural properties.     

正常肉和白肌肉的蛋白质变化

测定白肌肉(Pale,Soft,Exudative,PSE)和正常猪肉(Reddish-pink,Firm,Non-exudative)持水性指标和在宰后不同时间的蛋白溶解性和蛋白降解程度。结果显示:PSE肉的各种蛋白溶解性都显著低于正常肉(p<0.05),宰后4 h的总蛋白、肌浆蛋白和肌原纤维蛋白的溶解性与滴水损失和贮藏损失都呈显著负相关,与持水力呈显著正相关。正常肉和PSE肉的肌浆蛋白和肌原纤维蛋白的SDS-PAGE电泳条带存在差异,PSE肉早期伴肌动蛋白(Nebulin)含量明显低于正常肉。可见PSE肉肌原纤维的降解较快,宰后早期的蛋白溶解性可以用于预测猪肉的持水性。     

Effects of combining microbial transglutaminase and high pressure processing treatments on the mechanical properties of heat-induced gels prepared from arrowtooth flounder (Atheresthes stomias)

The objective of this study was to evaluate the effect of setting conditions (25 °C for 2 h or 40 °C for 30 min) and combining of microbial transglutaminase (MTGase) and high pressure processing (HPP) on the mechanical properties of heat induced gels obtained from paste from arrowtooth flounder (Atheresthes stomias). Treatments included fish paste control without added MTGase, fish paste incubated with MTGase but not pressurized (MTGase + cooking), fish paste incubated with MTGase and pressurized at 600 MPa for 5 min (MTGase + HPP + cooking) and fish paste pressurized at 600 MPa for 5 min and incubated with MTGase (HPP + MTGase + cooking). The controls and the treated samples were then subjected to one of two thermal treatments: 90 °C for 15 min or 60 °C for 30 min before cooking at 90 °C for 15 min. Samples of fish paste heated at 60 °C before cooking could not be used to prepare gels for texture profile analysis (TPA). TPA showed that pressurization improved the mechanical properties of gels made from paste treated with MTGase and set at 25 °C. The opposite was observed for samples set at 40 °C. Setting at 40 °C appeared to induce proteolytic degradation of myofibrillar proteins.     

Oxidation in HiOx-packaged pork Longissimus muscle predisposes myofibrillar and sarcoplasmic proteins to N-nitrosamine formation in nitrite-curing solution

The effect of meat protein in situ oxidation on the formation of N-nitrosodiethylamine (NDEA) was investigated. Fresh minced pork was untreated (Con) or treated with 700 mg/kg α-tocopherol (Toc) or 300 mg/kg tea polyphenols (PPE), packaged in a HiOx atmosphere (78.8% O₂, 18.8% CO₂, 2.4% N₂), then stored at 2 ± 1 °C for up to 10 days. Crude myofibrillar (MP) or sarcoplasmic (SP) protein (20 mg/mL) extracted from stored meat was reacted with 43 μM sodium nitrite at 80 °C for 1 h. Lipid oxidation was totally inhibited in PPE pork but increased in Con and Toc samples after 10 days. There was significant protein oxidation (losses of sulfhydryls, formation of protein carbonyls) in both MP and SP in all samples during storage. However, the Con group suffered more extensive protein oxidation than Toc and PPE and produced more NDEA (P < 0.05), indicating that protein oxidation promoted nitrosation.     

A proposed mechanism of tenderising post-rigor beef using high pressure–heat treatment

Tenderness of beef M. Sternomandibularis was tough when cooked from both raw, and when previously heated (60 °C, 20 min), whereas a significant improvement in tenderness was achieved when pressure–heat (P–H) treated muscle (200 MPa, 60 °C, 20 min) was cooked. In order to determine the mechanism for this improvement, connective tissue, myofibrillar and sarcoplasmic proteins, were separated into three fractions and studied with regard to their solubilisation, denaturation and aggregation, degradation and strengthening of protein structures for the three treatments (raw, heated and H–P treated). Measurements included DSC, SDS–PAGE, surface hydrophobicity, and the appearance, length and width of myofibres (light microscopy). For the connective tissue fraction, heat solubility was determined.     

Modification of pasta structure induced by high drying temperatures. Effects on the in vitro digestibility of protein and starch fractions and the potential allergenicity of protein hydrolysates

The effects of drying on pasta structure, starch and protein digestibility and potential allergenicity were investigated. Pasta was dried at low (55 °C, LT), high (70 °C, HT) and very high temperature (90 °C) applied either at the beginning (VHT) or at the end of the drying profile (VHT_LM). Changes in dried and in cooked pasta structure were determined regarding protein solubility, thermal properties of starch, microscopic and rheological measurements. Changes were moderate up to 70 °C and increased at higher temperatures and especially for VHT_LM drying.