Thermal Denaturation in Fish Muscle Proteins During Gelling: Effect of Spawning Condition

We studied thermal denaturation of myofibrillar proteins from pre-and post-spawning hake by differential scanning calorimetry (DSC), and evaluated denaturation kinetics under both conditions. The denaturation enthalpies of all pre-spawning fish muscle extracts were less than those from post-spawning. The area under the DSC thermogram corresponding to myosin denaturation was smaller in myofibrillar extracts from pre-spawning than from post-spawning hake, while the areas corresponding to denaturation of actin were similar. Between 40 and 55°C the myosin denaturation rates were greater for post-spawning than for pre-spawning hake. Both entalphies and kinetic data indicated proteins of fish in a better biological condition (post-spawned) denature more rapidly and completely.     

Thermal Denaturation and Aggregation of Proteins in Minced Fish as Studied by a Thermomechanical Method

A thermomechanical method of heating and mixing, was developed to study directly protein denaturation and aggregation in minced fish. It is based on simultaneous and continuous measurement of torque and temperature in a meat sample mixed while being heated. The torque and temperature were continuously recorded. From curves thus obtained, thermal denaturation temperatures of minced samples from 6 marine fish species were determined. The curves showed 4-6 peaks at 32-38°C, 44-46°C, 52-56°C, and 62-75°C, presumably corresponding to denaturation of myosin, actomyosin, sarcoplasmic proteins, and actin, respectively. The temperature range of peak 1 was 4-5°C higher in fatty fish (herring, mackerel) than in lean fish (cod, blue whiting).     

Scanning Calorimetric Behavior of Tilapia Myosin and Actin due to Processing of Muscle and Protein Purification

The thermal behavior of tilapia muscle proteins was investigated by differential scanning calorimetry at various stages in the processing of surimi and during purification of myosin and actin. A shift in the thermal transition of actin to lower temperature was observed and the enthalpies of denaturation for both actin and myosin decreased with further processing. Salt addition also induced shifts in denaturation transitions to lower temperatures and decreased enthalpies of denaturation.     

Species dependence of fish myosin stability to heat and frozen storage

The thermal stabilities of the proteins of a range of fish muscles of different habitat temperatures were determined by differential scanning calorimetry before and after frozen storage at 20°C.
In whole muscle a clear relationship was seen between habitat temperature and the thermal denaturation of myosin, which persisted when isolated myosins were analysed under conditions close to physiological pH and ionic strength. the ionic environment of the myosin molecules in the whole tissue will, however, not be exactly the same as in the myosin solution.
No significant correlations were seen between habitat temperature, ultimate pH and other analytical parameters.
After frozen storage, the myosin transitions in red snapper, a warm water species, was markedly changed in whole muscle but not in isolated myosin, suggesting the post mortem development of an interaction with other muscle proteins. In contrast, in cod, a cold water species, changes in myosin transitions were very similar, both in whole muscle and isolated myosin.
The implications of species differences in the 'melting' of myosin domains and changes in textural quality during frozen storage are discussed briefly.     

Relationship Between Thermal Denaturation of Porcine Muscle Proteins and Water-holding Capacity

ABSTRACT: Differential scanning calorimetry was used to investigate denaturation characteristics of pork muscle proteins from carriers and noncarriers of the RN-gene. Pork from RN-carriers deviated from noncarriers in maximum denaturation temperatures and denaturation enthalpy, with proteins of RN-carriers being the most heat-labile. Correlation studies on the results showed that water-holding capacity was significantly correlated to changes in enthalpy of the population mainly representing myosin tails and sarcoplasmic proteins (p < 0.001). Finally, the influence of ultimate pH and preheating on thermal characteristics of porcine muscle proteins was studied. Myosin tails and sarcoplasmic proteins were most sensitive to pH changes, while myosin heads were most sensitive to preheating simulating stress-induced temperature increases.     

Thermal stability of fish myofibrils: a differential scanning calorimetric study

The variation in myofibrillar protein thermostability was compared for various fish species, using differential scanning calorimetry. The tropical fish, catfish ( Clarius gariepinus ), carp ( Cyprinus carpio ), Nile perch ( Lates niloticus ), red snapper ( Lutianus sebae ), red mullet ( Parpeneus barberinus ), sea bream ( Gymnocranius rivalatus ), and cold-water reared trout ( Salmo gairdneri ) and cod ( Gadus morhua ) were analysed. Onset temperature of myofibrillar protein denaturation occurred at up to 11°C higher for tropical species (43.5°C, catfish), than cod (32.6°C) at pH 7 and low ionic strength (I). As pH (6.0-8.0) and I (0.05-1.00) were increased, thermal denaturation temperatures of myosins from tropical, but not cold-water, species decreased. Enthalpies of myofibrillar denaturation decreased for all species with increasing pH and I. Only one thermal transition was detected for myosin at pH 6 and low I, increasing to three as pH and I were increased. Changes in thermal characteristics of myosin subunits over iced and frozen storage suggest more rapid deterioration in cold-water than in tropical fish. The differences in myofibrillar stability of fish from different habitat temperatures have implications for the processing and storage of tropical fish.     

鱼肉蛋白的热变性研究进展

在鱼肉蛋白的加工过程中,加热处理是最常见的工序之一。热处理造成的蛋白热变性会对蛋白质的理化性质和功能特性产生很大的影响,且这种影响多难以复原。本文总结了近几年来国内外关于鱼肉蛋白在加热过程中蛋白组成成分、总巯基和羰基、溶解度、浊度、黏度、Ca2+-ATPase活性、凝胶、质构等特性指标的变化趋势的研究进展,结合蛋白质热变性的机理深入分析其变化原因,提出今后在鱼类加工方面对于鱼肉蛋白热变性的研发方向,旨在为鱼制品选择合理的热加工工艺的提供理论依据。     

Hydroxyl radical oxidation destabilizes subfragment-1 but not the rod of myosin in chicken myofibrils

The thermal denaturation process of myosin in oxidized chicken myofibrils was investigated. Exposures of myofibrils to hydroxyl radical-generation systems (HRGS) resulted in an enhanced susceptibility of myosin to thermal inactivation of Ca-ATPase and a loss of salt solubility. The chymotryptic production of subfragment-1 (S-1) from myosin in oxidized myofibrils decreased more rapidly than that in un-oxidized myofibrils upon heating, which paralleled the Ca-ATPase decay. However, the heat-induced decrease in chymotryptic production of rod from myosin was not affected by the HRGS treatment. The results suggested that free radical oxidation promoted thermal destabilization of myosin in the S-1 portion instead of the rod portion. The altered myosin denaturation pattern due to hydroxyl radical oxidation was likely a cause for functionality changes in oxidatively stressed myofibrillar proteins in meat processing.     

Differential scanning calorimetric study of muscle and its proteins: Myosin and its subfragments

The thermal denaturation of rabbit skeletal muscle myosin and its subfragments was investigated by differential scanning calorimetry. The thermal denaturation of myosin was shown to occur via three (at least) partly independent cooperative endothermic processes. The temperatures at which these processes occurred (312, 317 and 323 K at pH 6.0 and I=1.0) were shown to vary with pH (5.5–8.0) and I (0.05–1.0). The apparent enthalpy of denaturation of myosin was also shown to be dependent on pH and I. By comparing thermograms of myosin with those of the isolated myosin subfragments, the three major processes associated with the thermal denaturation of rabbit myosin could be tentatively assigned to different regions of the myosin molecule, namely, the helical tail, the ‘hinge-region’ and the globular heads. The ‘hinge-region’ thermal denaturation was shown to be reversible at pH 6.0 and I=1.0. Investigations of the effects of ortho-, pyro-, and tripolyphosphate on the thermal denaturation of myosin showed that added pyrophosphate destabilised the myosin molecule by about 9 K compared to the effects of ortho and tripolyphosphate, even though the latter was probably hydrolysed to ortho and diphosphate.     

Structural and Functional Changes in Myofibrillar Proteins of Sea Salmon (Pseudopercis semifasciata) by Interaction with Malonaldehyde (RI)

ABSTRACT: Changes on the myofibrillar proteins of Sea Salmon ( Pseudopercis semifasciata ) induced by malonaldehyde were investigated. Electrophoretic patterns, solubility, and differential scanning calorimetric studies were performed after incubation of proteins with malonaldehyde (MDA) at 27 °C. Results obtained showed a different thermal behavior, evidencing a decrease in thermal stability, changes in denaturation enthalpies values (ΔH_total and ΔH_myosln), and the appearance of new molecular species with a loss in the cooperativity of the myosin denaturation. A decrease in solubility and SDS-PAGE profiles revealed the participation of myosin and other proteins in the formation of aggregates involving nondisulfide covalent linkages for myosin heavy chain (MHC) and disulfide bridges in some other proteins.     

Cryoprotection Affects Physiochemical Attributes of Rainbow Trout Fillets

ABSTRACT: Stabilization of trout myofibrillar proteins during −20°C, 90-d storage and after soaking fillets in water, 8.0% sucrose/sorbitol, or 1.0% sodium lactate was investigated with or without 0.5% phosphate and with or without 0.05% MgCl₂. Fillets not soaked were the control. Compared with the control, cryoprotectants increased total protein and myofibrillar protein solubility; decreased surface hydrophobicity, total, free, and disulfide sulfhydryl content, and myosin susceptibility to thermal denaturation. There were no differences in total protein solubility and actin susceptibility to thermal denaturation between cryoprotectants and the water treatment. Phosphate minimized frozen storage effects on actin solubility and reduced protein surface hydrophobicity and myosin susceptibility to thermal denaturation, while MgCl₂ increased the negative effects of frozen storage.     

Analysis of the effects of thermal protein denaturation on the quality attributes of sous‐vide cooked tuna

Traditional (low temperature, long time) and novel (low temperature, short time) sous‐vide cooking of lean tuna were characterized by analyzing the effects of thermal protein denaturation (TPD) on quality attributes, such as color, appearance, shrinkage, drip loss, and texture. TPD was analyzed by differential scanning calorimetry and estimated for several thermal schedules by kinetic analysis, following the dynamic method. When heated at a rate of 10 °C/min, myosin began to denature at around 35 °C. Actin did not denature, even when the temperature rose to approximately 51 °C, until the denaturation of myosin was complete. However, actin began to denature at approximately 58 °C and was completely denatured at 76 °C. Actin denaturation had a stronger effect than myosin denaturation on texture changes, whereas myosin denaturation was responsible for changes in color and appearance. A better preservation of tuna quality was obtained by novel sous‐vide cooking over the traditional sous‐vide method.

Practical applications

The results of this study are useful to both the research community and industry because they provide quantitative characterization of the consequences of sous‐vide cooking method on food quality explained by estimating TPD. Moreover, the kinetic parameters of the denaturation rate collected for kinetic modeling of the TPD of tuna, not only have application to simulate denaturation of actin and myosin under different thermal schedules of sous‐vide cooking, but also they can be used for the analysis of additional thermal treatments.     

TEXTURE AND COLOUR CHANGES IN MEAT DURING COOKING RELATED TO THERMAL DENATURATION OF MUSCLE PROTEINS1

Differential scanning calorimetry was used to select temperatures that diferentiated between thermal denaturation of the three major structural protein species in bovine muscle: myosin, collagen and actin. Samples of m. semimembranosus, m. semitendinosus and m. psoas major were heated to those different temperatures and evaluated sensorially. Three groups of sensory properties were needed to describe the main texture changes observed in the meat: 1. Firmness 2. Fiber cohesivity, bite-off force, residual bolus 3. Juiciness Firmness increased with thermal denaturation of the myofibrillar proteins (myosin; 40-60d?C and actin; 66-73d?C). Fiber cohesivity etc. decreased with collagen denaturation (56-62d?C). Reduction in juiciness was primarily associated with actin denaturation, while cooking loss increased over the whole temperature range. The property “total chewing work”, a composite of the first two texture groups mentioned, yielded like the judges' “total texture preference”, optimal texture in the 60-67d?C temperature region, implying denatured myosin and collagen but native actin. This meat was light pink-gray in colour, while the cooking juice released was dark red.     

NMR relaxometry and differential scanning calorimetry during meat cooking

By combining simultaneous nuclear magnetic resonance (NMR) T₂ relaxometry and differential scanning calorimetry (DSC) on pork samples heated to nine temperature levels between 25 and 75 °C, the present study investigates the relationship between thermal denaturation of meat proteins and heat-induced changes in water characteristics. Principal component analysis (PCA) on the distributed ¹H NMR T₂ relaxation data revealed that the major changes in water characteristics during heating occur between 40 and 50 °C. This is probably initiated by denaturation of myosin heads, which however, could not be detected in the DSC thermograms obtained directly on the meat. In contrast, the DSC thermograms revealed endothermic transitions at 54, 65 and 77 °C, probably reflecting the denaturation of myosin (rods and light chain), sarcoplasmic proteins together with collagen and actin, respectively. Simultaneous modelling of DSC and NMR data by partial least squares regression (PLSR) revealed a correlation between denaturation of myosin rods and light chains at 53–58 °C and heat-induced changes in myofibrillar water (T₂ relaxation time 10–60 ms) as well as between actin denaturation at 80–82 °C and expulsion of water from the meat. Accordingly, the present study demonstrates a direct relationship between thermal denaturation of specific proteins/protein structures and heat-induced changes in water mobility during heating of pork.     

5 种淡水鱼肌肉热特性比较研究

为研究不同品种名优淡水鱼冷藏保鲜及热加工的特性,采用差示扫描量热法研究武昌鱼、草鱼、黄颡鱼、鲈鱼和鳜鱼5 种新鲜淡水鱼鱼肉的水分含量、冰点、变性温度、变性热焓和比热容并进行品种间的比较。结果发现：5 种淡水鱼冰点分布在－0.57～－0.1 ℃之间;50 ℃附近热吸收峰起始温度分布在37～41.57 ℃之间,终止温度分布在60.47～62.33 ℃之间;变性热焓在1.680 0～2.499 7 J/g之间;70 ℃附近热吸收峰的起始温度分布在66.33～71.13 ℃之间,终止温度分布在73.60～82.20 ℃之间,变性热焓在0.418 0～0.512 2 J/g之间;鱼肉的比热容在1.658 0～3.862 3 J/（g·K）之间。结果表明：在5 种新鲜淡水鱼中,武昌鱼水分含量和冰点相对较低;草鱼肌球蛋白和鳜鱼肌动蛋白变性温度和变性热焓相对较高,蛋白质稳定性更高;新鲜鱼肉的比热容高于冻结鱼肉,加工后鱼肉的比热容高于未加工鱼肉。     

Myosin thermal stability in fish muscle

The thermal stability of fish muscle proteins varies between species with considerable implications for storage and processing properties. Myosins were isolated from cod and snapper, cold- and tropical-water fish respectively, and their thermal melting characteristics were compared using differential scanning calorimetry over a range of pH and ionic strength conditions. At pH 6 and ionic strength 0.06 M, cod myosin exhibited a thermal melting transition 10 K lower than snapper myosin, reflecting a comparable differential in transitions attributed to myosin in intact muscle from the two species at 315 and 325 K, respectively. However, with increasing pH and ionic strength, conditions favourable to disaggregation and dissolution of myosin in vitro, the snapper myosin transition decreased to that exhibited by cod myosin, which remained essentially unaffected by the conditions. It is suggested that differences in the aggregation characteristics of fish myosins may explain in part the different thermal melting properties in vitro.     

Extractability and thermal stability of frozen hake (Merluccius merluccius) fillets stored at −10 and −30 °C

The relationship between thermal stability changes and functionality loss was monitored in hake muscle fillets stored for 40 weeks at ?10 and ?30 °C. The evolution of changes in apparent viscosity, dimethylamine formation and extractability of muscle proteins in NaCl, sodium dodecyl sulphate (SDS) or SDS plus mercaptoethanol showed drastic differences as a function of temperature. At the higher storage temperature, both myosin heavy chain and collagen were the most severely unextracted in salt and SDS solutions, with actin becoming unextractable at the end of storage. Differential scanning calorimetry showed differences with storage time and temperature in both onset temperature and thermal denaturation enthalpy, mostly affecting the myosin transitions. Some protein denaturation occurred with little or no functionality loss. A considerably high fraction of hake muscle proteins remained in the native‐like condition even at the higher frozen storage temperature. In these conditions both apparent viscosity and myosin and actin extractability in NaCl were very low. © 2002 Society of Chemical Industry     

Effects of freezing and thawing methods and storage time on thermal properties of freshwater prawns (Macrobrachium rosenbergii)

Thermal stability of proteins in frozen and thawed freshwater prawns was measured by differential scanning calorimetry. The onset and peak melting temperatures corresponding to myosin denaturation, as well as total enthalpy of denaturation of prawn muscle, decreased after freezing–thawing treatments. There were no significant differences in the thermal properties of prawns with changes in the rate of freezing, ie blast (fast) vs still (slow) freezing. However, the thermal properties were influenced by the rate of thawing. Rapid thawing using a combination of microwave and tap water resulted in a lower (P⩽0·05) thermal stability of prawn proteins compared to slow (refrigeration) or moderately fast (tap water) thawing methods. Keeping prawn shells intact or not intact during freezing–thawing did not alter the thermal properties of the prawn proteins. © 1997 SCI.     

Thermal Properties of Proteins in Chicken Broiler Tissues

The thermal behavior of breast and thigh muscles, blood and skin tissues of chicken broilers was evaluated by differential scanning calorimetry (DSC). Onset temperature of transition (T_o), maximum thermal transition (T_max) temperatures, and denaturation enthalpy (ΔH) were evaluated. Breast muscle exhibited a complex thermogram with five endothermic transitions at 57°C, 63°C, 67°C, 73°C, and 78°C at a heating rate of 10°C/min. Thigh muscle exhibited only three major transitions at 60°C, 66°C, and 76°C. Thermal curves of isolated protein fractions indicated that the thermal transitions in muscle corresponded to the denaturation of myosin, sarcoplasmic proteins, collagen and F-actin. An increase in the heating rate from 1.0° to 40°C/min significantly elevated the onset temperature of transition and major transition temperatures, as well as the enthalpy of denaturation. Enthalpy of the muscle system heated to various end-point temperatures, cooled and reheated, showed that myosin was completely denatured at 60°C, sarcoplasmic proteins at 70°C and actin at 80°C.     

Suppression of thermal denaturation of myosin and salt-induced denaturation of actin by sodium citrate in carp (Cyprinus carpio)

The effect of sodium citrate (Na-citrate) on myosin and actin denaturation in myofibrils was investigated. Na-citrate significantly suppressed the thermal inactivation of Ca²⁺-ATPase of carp myosin in a concentration-dependent manner. The effect was greater than that of sorbitol. A similar effect was observed with myofibrils in which myosin is stabilized by F-actin binding. Na-citrate dissolved myofibrils at lower concentration than NaCl. Nevertheless, Na-citrate at 1 M failed to denature F-actin in myofibrils, while 1 M NaCl denatured F-actin almost completely. Na-citrate suppressed the NaCl-induced F-actin denaturation. Sorbitol did not show such protective effect on F-actin denaturation. Moreover, Na-citrate suppressed the freeze denaturation of myofibrils at lower concentration than sorbitol. Thus, Na-citrate was proved to be superior to sorbitol. It was suggested that Na-citrate alone could substitute sorbitol as cryoprotectant in surimi and NaCl as dissolving reagent of myofibril in thermal gel production.