Thermal denaturation of proteins in Post rigor muscle tissue as studied by differential scanning calorimetry

Post rigor bovine M. semimembranosus was analysed by differential scanning calorimetry (d.s.c.). After extractive removal of sarcoplasmic proteins, subsequent pH adjustment and manual connective tissue removal, d.s.c. yielded reproducible thermograms which permitted investigation of the individual major myofibrillar proteins in various pH and salt environments without prior isolation. The positions of two major peaks, interpreted as myosin transitions, proved to be strongly pH dependent. At pH 5.4, the peak maxima occurred at 58 and 65°C, respectively, at a heating rate of 10°C min-¹. Above pH 6.5 their order of denaturation was reversed. In the pH range 5.4–6.5 the peak ascribed to actin had its maximum near 80°C in intact muscle. Above this pH range it was displaced to lower temperature. The thermal stability of actin was studied after treatment of the muscle tissue with different salt solutions. At equal ionic strengths (μ = 0.15) at pH 5.5, calcium chloride and sodium chloride caused 6.5°C and 4°C displacement to lower temperature, respectively. The thermograms of bovine semimembranosus muscle were compared to those of two red and two white muscle types (bovine cardiac and rabbit soleus muscles, chicken breast and rabbit semimembranosus muscles, respectively) at two pH levels. Greater myosin differences were found between red and white muscles than between muscles from different animal species. All muscles gave similar actin transitions, with exception of the heart muscle where the actin peak appeared at 3 °C lower temperature. The necessity of a strict pH control in order to obtain reproducible muscle thermograms is demonstrated.     

Gelation characteristics of muscle proteins from pale, soft, exudative (PSE) pork

Thermally induced protein gels were made by using extracted salt-soluble proteins from normal pigs and stress-susceptible pigs determined to have pale, soft, exudative (PSE) muscle. Effects of heating rates (17,39 and 93° C/h) at various protein concentrations (23, 34, 48 and 54 mg/ml) were evaluated. Gel strength of PSE extracts was 45% of the controls at equivalent protein concentration. Gel strength of normal and PSE-muscle protein gels from the first compression curve increased with increasing protein concentrations at all heating rates; however, gel strength was greater for slow heating rates than for fast heating rates in both PSE and normal samples. Percent water loss was greater for PSE extracts than for controls at the same protein concentration. Losses of 47% and 36% for PSE and controls, respectively, were observed at a a protein level of 54 mg/ml protein. There was no heating rate effect on water losses in either case. Protein loss was less, for both PSE and control, at low protein concentrations than at high protein content in the range studied. Slow heating rates resulted in less protein loss, for both PSE and control, whereas faster heating rates gave greater protein losses in the exuded water.     

辐照处理对PSE肉的物化、微生物及感官特性影响(英文)

在屠宰厂与农贸市场肉品卫生检验中,常发现以肉色苍白、肉质柔软和切面多汁为特点的生理异常肉,中国称之为"白肌肉"(PSE肉)。PSE肉外观色泽苍白,适口性差,易腐败,因而不受消费者欢迎。为了提高PSE肉的外观颜色及微生物安全性,并考虑因辐照产生的异味及微生物杀菌效果,选择0和4.5kGy的辐照剂量对PSE肉进行处理。之后在4℃下储藏2周,并测定pH、颜色、脂肪过氧化值、微生物安全性及官能特性等指标。结果表明:辐照在改善PSE肉的微生物安全性的同时,还能改善PSE肉色。     

Protease activity and the ultrastructure of broiler chicken PSE (pale,soft, exudative) meat

The biological cause of broiler PSE meat seems to be an excessive release of Ca²⁺, promoted by a genetic mutation of ryanodine receptors located in the sarcoplasmic reticulum of skeletal muscle cells. Excessive Ca²⁺, associated with protein denaturation in meat, enhances protease activity and influences the functional properties of PSE meat. Twenty-four-hour post-mortemPectoralis major m. samples exhibited lower values for pH, water-holding capacity, and shear force than did control samples, in contrast to colour (L∗) and cooking loss values. Protease activity, measured as myofibril fragmentation index, presented higher values in PSE meat than in control samples. Ultrastructural examination revealed shrinking and depolymerisation of myofilaments and Z-lines disorganisation within the sarcomere in PSE meat. Intense calpain activity was also observed, indicating that the process may initiate at the filaments, because of protein denaturation, and spread through Z-lines, resulting in the collapse of the sarcomere structure.     

Progress in reducing the pale, soft and exudative (PSE) problem in pork and poultry meat

Research in the area of the pale, soft and exudative (PSE) pork and poultry meat is reviewed in this article with an emphasis on genetic, biochemical and metabolic factors contributing to the problem. Over the past five decades, there has been much more work in the pork meat area where a few genetic markers have been identified, and are currently used to remove susceptible animals from the herd. Some of the markers are linked to aberrant calcium regulation in the early postmortem muscle. The poultry industry is still not at the point of using genetic marker(s); however, some recent work has revealed several potential markers. The review also discusses environmental factors such as antemortem stress and early postmortem processing practices (e.g. chilling rate) that can influence the development and severity of the PSE phenomenon. Some of these factors are known to cause protein denaturation at the early stage of postmortem and directly contribute to poor water-holding capacity and inferior texture in fresh meat and later in processed products. A newer hypothesis suggesting that variation in protein oxidation, in response to antemortem stress and early postmortem tissue environment, can contribute to development of PSE pork is also discussed. Finally, a few recommendations for future work are proposed.     

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.     

Beef muscle osmotic pressure as assessed by differential scanning calorimetry (DSC)

Osmotic pressure, an important physiological characteristic of plant and animal tissues, is difficult to measure, and no convenient and efficient method is so far available. In the present work we developed a differential scanning calorimetry (DSC) method, and estimated the physiological osmotic pressure of intact bovine muscle and post-mortem variation during the conversion of muscle into meat. We also examined the major role of sample pH in this variation. This method, based on the determination of the melting onset point of frozen muscle's indigenous water, was compared to a freezing point osmometric method and a soaking method. It was concluded that the DSC method appeared more practical and reliable than other methods and could be applied to various animal and plant tissues and fluids.     

Electrostatic effects on β-lactoglobulin transitions during heat denaturation as studied by differential scanning calorimetry

This work comprises the study of the thermal treatment of β-lg and its denaturation as a function of pH and ionic strength followed by differential scanning calorimetry. The concentration of protein was 14 (w/v)% in order to study the behaviour of highly concentrated β-lg solutions during heating. The denaturation temperature of β-lg was dependent on both pH and ionic strength, meaning that electrostatic interactions between protein monomers in the native state were important for the denaturation of β-lg. The thermograms from the calorimetric measurements also revealed that the quarternary structure of β-lg at pH-values close to the isoelectric point was influenced by the presence of salt and the nature of the salt (NaCl, KI and LiI). Small exotherms emerged in the thermograms at the low temperature side of the denaturation temperature for β-lg. The presences of these exotherms are probably caused by restructuring of the quarternary structure of native β-lg prior to denaturation, due to dissociation into smaller entities and possible also formation of a liquid crystalline-like structure in the highly concentrated protein solution. The present study provides a contribution to the understanding of the importance of the electrostatic interactions between native β-lg molecules and how different salts and ionic strengths affect the denaturation properties of the protein in concentrated systems.     

Application of high pressure processing to improve the functional properties of pale, soft, and exudative (PSE)-like turkey meat

Improvement of functional and rheological properties of turkey breast meat proteins with different ultimate pHs at 24 h post-mortem (pH₂₄) was attempted using high pressure processing (up to 200 MPa for 5 min at 4 °C). Pressures of 50 and 100 MPa were found to increase the water holding capacity of low pH meat. At these pressures, higher protein surface hydrophobicity and greater exposure of sulfhydryl groups were evident. These elements may have contributed to improved water retention properties of the treated protein. The formation of a better gel network was also evident at 50 and 100 MPa as revealed by the dynamic viscoelastic behavior. Application of high pressure significantly (P < 0.05) increased total protein solubility in both low and normal pH meats. Aggregation of myofibrillar proteins increased in low pH meat at higher pressure (200 MPa) as revealed by SDS-PAGE profile.

Industrial relevance

A major concern in the poultry industry is reduced meat functionality, such as low water holding capacity (WHC) in low pH poultry meat leading to reduced yield causing economic loss in the production of further processed products. An alternative technology to reduce salt and improve water retention properties is by the application of high pressure processing (HPP) to produce healthier food products.     

Volatile production in irradiated normal, pale soft exudative (PSE) and dark firm dry (DFD) pork under different packaging and storage conditions

Normal, pale soft exudative (PSE) and dark firm dry (DFD) pork Longissimus dorsi muscles were vacuum packaged, irradiated at 0 or 4.5 kGy and stored at 4°C for 10 days. Volatile production from pork loins was determined at Day 0 and Day 10 of storage at 4°C. With both aerobic and vacuum packaging, irradiation increased the production of sulfur-containing volatiles (carbon disulfide, mercaptomethane, dimethyl sulfide, methyl thioacetate and dimethyl disulfide) in all three pork conditions at Day 0 but did not increase hexanal – the major indicator volatile of lipid oxidation. The PSE pork produced the lowest amount of total sulfur-containing volatiles in both aerobically and vacuum-packaged pork at Day 0. The majority of sulfur-containing volatiles produced in meat by irradiation disappeared during the 10-day storage period under aerobic packaging conditions. With vacuum packaging, however, all the volatiles produced by irradiation remained in the packaging bag during storage. Irradiation had no relationship with lipid oxidation-related volatiles (e.g. hexanal) in both aerobic and vacuum-packaged raw pork. The DFD muscle was very stable and resistant to oxidative changes in both irradiated and nonirradiated pork during storage, suggesting that irradiation can significantly increase the utilization of raw DFD pork and greatly benefit the pork industry.     

Preslaughter resting and hot-fat trimming effects on the incidence of pale, soft and exudative (PSE) pork and ham processing characteristics

Effects of preslaughter rest (0, 1, 2 or 3 h) and hot-fat trimming (HFT, one side) on muscle quality characteristics of hams and loins were tested with 120 pigs. Center-cut loins from each carcass side were vacuum packaged and stored for 7 or 14 days at 2°C. Pigs not rested before slaughter had a skin temperature 3°C higher and ham muscle temperatures 1°C higher than pigs rested 1-3 h (P<0.05). Three hour muscle pH was highest if pigs were rested for 1 h. HFT significantly accelerated carcass chilling rate, but did not affect rate of muscle pH decline. After 14 days of aging, pigs not rested had lower sensory panel ratings for juiciness, tenderness, flavor and overall mouthfeel of loin chops than pigs rested 1 or 2 h. Percentage bound water of hams was highest (P<0.05) if pigs rested 1 or 2 h rather than a shorter or longer times. Pigs should be rested 1 h before slaughter and HFT carried out to allow more rapid carcass chilling and improve muscle quality.     

Effect of oral loading of acid or base on the incidence of pale soft exudative pork (PSE) in stress-susceptible pigs

Thirty-three halothane-positive pigs were assigned to one of three ad libitum water treatments 4 days prior to slaughter: control, sodium bicarbonate (12·6 g litre(-1)), or ammonium chloride (8 g litre(-1)) to study the effect of an oral acid or base load on post-mortem changes in the longissimus dorsi muscle and on ultimate pork quality. Eight halothane-negative pigs were also slaughtered as negative controls. Dietary ammonium chloride caused a metabolic acidosis as indicated by lower blood and urine pH, and lower blood bicarbonate in treated pigs. In halothane-positive pigs, dietary treatments had no significant effect on post-mortem muscle pH changes or on measurements of muscle quality (colour, drip loss, texture scores or protein solubility). There was a weak to modest relationship between the blood gas measurements and meat quality indicators. Blood pH (r = 0·566) and bicarbonate (r = 0·514) were positively correlated with subjective texture scores and a trend was seen for blood PCO(2) and bicarbonate to be positively related to 45 min pH and subjective colour. These observations suggest that pigs with increased blood pH and bicarbonate may show an improvement in meat quality. Further research is necessary to find the optimum time and level of treatment to maximize the response.     

Physicochemical and structural properties of myofibrillar proteins isolated from pale,soft, exudative (PSE)-like chicken breast meat: Effects of pulsed electric field (PEF)

The present study was performed to clarify the effects of pulsed electric field (PEF) on the physicochemical properties and conformations of myofibrillar proteins (MPs) extracted from pale, soft, exudative (PSE)-like chicken breast meat. Various PEF parameters, including the electric field intensity (0–28 kV/cm) and pulse frequency (0–1000 Hz), were varied. The results showed that as the PEF intensity increased, the solubility, surface hydrophobicity and sulfhydryl group content of the MPs were significantly improved. However, when the intensity exceeded 18 kV/cm, these properties declined, which was possibly due to protein aggregation caused by functional group interactions. Additionally, the PEF treatment altered the rheological properties of PSE-like MPs and induced the formation of low-elasticity MPs. However, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that the primary structure of the MPs was not altered after the PEF treatment. According to the circular dichroism (CD) spectroscopy results, the α-helix contents of the PEF-treated samples were increased but the β-turn and random coil contents were reduced.Industrial relevanceThe occurrence of PSE-like chicken meat is one of the most serious quality issues worldwide and can decrease the consumer's purchasing desire and result in extensive economic losses for the poultry processing industry. MPs play a vital role in the qualitative characteristics of chicken products. As a potential novel processing technology, PEF has potential applications for improving the functionality of PSE-like chicken protein to expand its application in the food industry.     

Chicken breast quality – normal,pale, soft and exudative (PSE) and woody – influences the functional properties of meat batters

Three classes (normal, PSE‐like and woody, thirty for per class) of fresh chicken breast meats were selected from a Chinese processing plant. Then, three classes of chicken breast meats were used to prepare meat batters and meatballs. This study investigated the effects of three classes of chicken breast meat on the physicochemical properties, water distribution, protein secondary structures and microstructures of meatballs. PSE‐like and woody meatballs both had lower water holding capacity and textural properties than normal meatballs. Furthermore, the free water mobility and proportion in PSE‐like and woody meatballs were increased (P < 0.05). According to near‐infrared spectroscopy results, the intensity of the 1940‐nm bands of PSE‐like and woody classes both increased. Both PSE‐like and woody meatballs formed more aggregated gel matrices than normal class. PSE‐like and woody classes had higher α‐helix and lower β‐structure contents (P < 0.05). Overall, compared with normal meatballs, PSE‐like and woody meatballs showed inferior functional properties.     

Effects of transportation time,distance, stocking density,temperature and lairage time on incidences of pale soft exudative (PSE) and the physico-chemical characteristics of pork

The study determined the effects of transportation time, distance, stocking density, temperature and lairage time on incidences of PSE and pork quality. Frequencies of PSE cases in stocking density categories within transport duration classes were determined. General linear models, regression and the principal component (PC) analysis were used to analyse the data. Highest incidences of PSE were recorded in autumn season while lowest incidences were recorded in the spring season. Transportation time and stocking density significantly affected pH_u and ultimately PSE incidences although there were no interactive effects. Highest risks of PSE occurrence were observed with more space allowance. The highest incidences of PSE were observed for animals that had travelled for two hours while the PSE cases were lower in animals that travelled for longer times. Distance travelled and transportation time had significant effects (P < 0.05) on thawing loss (TL) % of pork. No relationships were reported between the other pre-slaughter variables and pork quality attributes. With the exception of transportation time and distance travelled which had a positive relationship with TL%, variation in other pre-slaughter variables did not affect meat quality variables. The risks of PSE occurrence were dependent on stocking density and transportation time.     

Improvement of the physicochemical properties of pale soft and exudative (PSE) pork meat products with an extract from mechanically deboned turkey meat (MDTM)

Loss of protein functionality of pale soft and exudative (PSE) meat and a surplus of mechanically deboned turkey meat (MDTM) has negative economic impacts on the meat industry. The objective of this research was to add functional proteins from an extract of MDTM to PSE meat to create a value-added pork sausage product. Sausages manufactured from PSE incorporating an MDTM extract exhibited a 30% increase in rigidity (P<0.05) compared to those manufactured using brine alone. Sausages made from PSE and normal pork with the MDTM extract demonstrated little difference in water holding capacity (P>0.05), and reductions of 4.1 and 3.1% in cooking loss (P<0.05), respectively, compared to sausages made employing brine alone. Introducing functional proteins derived from MDTM to processed meat made with PSE pork has the potential to enhance the economic value to both of these low value raw materials.     

The effect of ionic strength, polyphosphates type, pH, cooking temperature and preblending on the functional properties of normal and pale, soft, exudative (PSE) pork

Compared to processed meat product made from normal pork, products made from pale soft exudative (PSE) pork have higher cook loss (CL) and weaker texture. In this study interactions between a range of processing conditions (ionic strength, polyphosphate addition, polyphosphate chain length, pH, cooking temperature and time between preparation and cooking), and their effect on the texture [shear stress (SS), true shear strain (TSS)] and CL of gels made from normal and PSE pork were examined. Of the processing conditions studied, ionic strength, polyphosphate addition and polyphosphate chain length affected the functional properties of normal and PSE pork differently. Generally, the functional properties of normal pork were superior to PSE pork, with no combination of conditions making all the functional properties of PSE pork equal to those of normal pork under the same conditions. The combination of conditions that was most effective in reducing the difference between normal and PSE pork was high ionic strength in the presence of added polyphosphate. Under these conditions there was no significant difference in CL between normal and PSE pork, although the texture (SS and TSS) of the PSE pork samples was still inferior.     

Thermal analysis of flaxseed (Linum usitatissimum) proteins by differential scanning calorimetry

The thermal properties of flaxseed (whole or dehulled), dehulled and delipidated meal, as well as proteins extracted and isolated from the meal, were investigated by differential scanning calorimetry (DSC). A high denaturation temperature (T_d) of 114.7 °C was observed for the major fraction of flaxseed protein isolated by anion-exchange chromatography, representing the 11–12 S storage globulin. Marked decreases in T_d and enthalpy were observed at pH 3 compared with pH of 5 or higher, while the presence of high salt (1.0 M NaCl) resulted in higher thermal stability, enthalpy and greater cooperativity of the transition. Thermal analysis of the major fraction in the presence of chaotropic salts or protein structure perturbants (sodium dodecyl sulfate, urea, dithiothreitol, N-ethylmaleimide, ethylene glycol) suggested the contributions of hydrophobic and ionic interactions, hydrogen bonding as well as disulfide linkages or disulfide–sulfhydryl (SS–SH) interactions, to the thermal stability of flaxseed protein.     

Methods of predicting pale, soft, exudative pork and their application in breeding programmes-A review

The condition of pale, soft, exudative (PSE) pork has long been considered to be mainly a post mortem phenomenon. There is now substantial evidence that such pigs are suffering from a kind of myopathy, which predisposes them to an abnormal post mortem metabolism. Genetic studies on PSE muscle indicate a moderate heritability for various post mortem muscle quality traits. Reliable methods for determining the abnormal condition in the live animal would make it possible to select more effectively and economically against stress and PSE-susceptibility. Three possible methods are: (1) Analysis of blood serum for CPK, aldolase, GOT or other enzyme activities with and without preceding exercise; and for blood groups. (2) Muscle biopsy analysis for glucose-6-phosphate, lactate or energy-rich phosphates. (3) Non-destructive testing of young pigs for sensitivity to the Malignant Hyperthermia Syndrome by allowing them to inhale the anaesthetic halothane (fluothane) for a 5 min period. The development of muscular rigidity and stiffness indicates a susceptibility to stress and a potential for PSE meat. The relationships of the various methods to ultimate muscle and carcass quality, as well as the problems inherent to each method, are discussed. It is concluded that the third test seems to be the most promising for application in the breeding of pigs for optimal stress resistance and muscle quality.     

Influence of weak organic acids and salts on the denaturation characteristics of intramuscular connective tissue. A differential scanning calorimetry study

Intramuscular connective tissue obtained from Longissimus dorsi muscle of a 4-year-old beef carcass was treated with NaCl solutions of 2, 4, and 6% (w/v), and CaCl₂ solutions of 50, 100, and 150 mM, and citric and lactic acid solutions of 0.5, 1.0, and 1.5% for three different marinating periods (24, 46, and 72 h). Changes in denaturation characteristics were investigated using differential scanning calorimetry and it was found that the denaturation onset temperature (T_o) and the denaturation peak temperature (T_p) increased as the NaCl concentration increased but decreased as the CaCl₂ concentration increased, irrespective of marinating time. Lactic and citric acid decreased T_o to about 39°C, from over 60°C breaking the structure of fibrils.