Gelling Properties of Actomyosin from Pre- and Post-Spawning Hake (Merluccius hubbsi)

The biochemical properties and the characteristics of heat-induced gelation of natural actomyosin (NAM) from pre- and post-spawning hake were studied. Mg²⁺ ATPase activity, reduced viscosity and myosin/actin mole ratio of NAM from post-spawning fish were higher than those of pre-spawning ones. Gelation of both actomyosin at 10 mg mL^?1 of protein concentration was optimal at 60°C and pH 6.0. The highest rigidity was reached at 0.40M and 0.44M KCl with NAM from pre and post-spawning hake, respectively. Irrespective of heating temperature, ionic strength conditions and at pH range 5.5–7.5, rigidity of post-spawning hake NAM gels was higher than those of pre-spawning fish. Scanning electron micrographs of pre- and post-spawning hake NAM showed “actin-type” and “myosin-type” ultrastructures, respectively.     

Cross-Linking of Myosin Heavy Chains from Cod, Herring and Silver Hake During Thermal Setting

Cross-linking of myofibrillar proteins extracted from cod (Gadus morhua), herring (Clupea harengus) and silver hake (Merluccius bilinearis) was studied in 0.6M NaCl, pH 6.5 at 40°C and evaluated turbidimetrically and by SDS polyacrylamide gel electrophoresis coupled with l-ethyl-3-(3-dimethylaminopropyl) carbodiimide as a zero-length crosslinker. Turbidities of heat-treated cod and silver hake myofibril/myosin solutions were significantly higher than those of herring. Electrophoretic results showed that the myosin heavy chain (MHC) was the principal myofibrillar protein cross-linked to form a polymerized complex during the heat treatment. Cross-linking ability of MHC from the three fish species was different; herring MHC formed only small polymers (n≦3) but cod and silver hake MHC formed both small and large polymers (n≦6).     

Mathematical modeling of the heat transfer process and protein denaturation during the thermal treatment of Patagonian marine crabs

Southern Ocean swimming crab Ovalipes trimaculatus and the Patagonian stone crab Platyxanthus patagonicus are fishing resources with commercial value. Thermal treatment of crabs is necessary to denature muscle proteins, facilitating meat detachment from the crab shell (picking procedure). The proximal composition, protein patterns of crab muscle, thermophysical properties and heat transfer coefficients were determined. Heat transfer during thermal processing of body (i.e., cephalothorax) and claws of both crab species was simulated using a finite element computational code; the simulations were experimentally validated. Color changes in crab muscle during the heating process were measured. Thermal denaturation kinetics of myofibrillar proteins was determined using Differential Scanning Calorimetry (DSC) in small samples previously heated in water under controlled conditions. DSC thermograms of raw crab muscle showed two peaks at 49.0 ± 0.4 and 77.5 ± 0.6 °C corresponding to myosin and actin respectively. Activation energies for the denaturation of myosin (145.70 kJ/mol) and actin (156.42 kJ/mol) were calculated from Arrhenius equation. The degree of denaturation achieved by the myofibrillar proteins at the coldest point of the muscle in body and claws during the heating process was established by considering the protein denaturation kinetics determined by DSC, the activation energies and the heat penetration curves. Adequate conditions for the detachment of meat from the crab exoskeleton were established. The obtained results may help in determining the optimal heating times during the industrialization of these crustaceans.     

Differential Scanning Calorimetry of Fish Muscle: The Effect of Processing and Species Variation

Differential Scanning Calorimetry (DSC) has been used to study the thermal properties of fish muscle proteins and to measure the extent of their denaturation under various processing conditions. Fish myosin was susceptible to denaturation by frozen storage and dehydration. Denaturation of certain fish proteins was partially reversible. Although fish myosin was very unstable, its thermal stability was found to increase in species adapted, to higher environmental temperatures.     

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.     

Effect of freezing rate on the denaturation of myofibrillar proteins

Freezing of bovine muscle has a denaturating effect on myofibrillar proteins, Differ-ential Scanning Calorimetry (DSC) studies of fresh and frozen muscle at different freezing rates show a decrease on denaturation enthalpies; the lower the freezing rate the greater the loss. When measuring the specific areas (ratio between each partial area in cm', and the dry weight of the sample, in mg) of the DSC thermograms, it can be observed that the area ascribed to myosin decreases with freezing, while the area corresponding to actin is not affected. These results are in agreement with the ATPase activity decreases as a consequence of freezing observing higher losses at lower freezing rates. The denaturation observed could be a result of a partial unfolding of the myosin head being more pronounced at low freezing rate.     

Changes in myofibrillar proteins during processing of pastirma (Turkish dry meat product) produced with commercial starter cultures

The effects of different commercial starter cultures (Staphylococcus carnosus, S. carnosus + Lactobacillus pentosus and Staphylococcus xylosus + Lactobacillus sakei), on myofibrillar proteins were investigated using differential scanning calorimetry (DSC) during the processing of pastirma. The stage of pastirma production significantly decreased the thermal stabilities of myosin and actin. Actin was less affected than myosin. The myofibrillar fraction of pastirma was hardly denaturated by S. carnosus, but more pronounced denaturation was obtained with S. carnosus + L. pentosus.     

Thermal Denaturation of Hake (Merluccius hubbsi) Myofibrillar Proteins. A Differential Scanning Calorimetric and Electrophoretic Study

Denaturation of hake (Merluccius hubbsi) proteins was studied with DSC by monitoring T_max of transitions and denaturation enthalpies. Whole muscle free of connective tissue showed two transitions (T_max 46.5°C and 75.3°C), and ΔH of 4.27 cal/g. The exudative sarcoplasmic fraction showed three transitions (T_max 45.2°C, 59.0°C and 75.5°C) and ΔH of 3.92 cal/g. The sarcoplasmic proteins from whole hake muscle contributed to both denaturation peaks. Muscle depleted of sarcoplasmic proteins by chloride extraction showed a higher thermal sensitivity and a diminished denaturation enthalpy on the second transition. This suggested an additional effect of chloride upon actin in addition to sarcoplasmic protein extraction. pH had an effect upon the native conformation of thick filament proteins, specifically myosin.     

Biochemical and Functional Properties of Myofibrils from Preand Post-Spawned Hake (Merluccius hubbsi Marini) Stored on Ice

Enzymatic activities assayed at beginning of storage in myofibrils from post-spawned hake were 3 X those in myofibrils from pre-spawned hake. Ca²⁺ sensitivity of myofibrils from pre-spawned hake was 40% less than that of myofibrils from post-spawned hake. The profiles of SDS-PAGE gels of pre-spawned myofibrils at beginning of storage showed a partially denatured myosin heavy chain, and polypeptide bands under myosin heavy chain. They probably represent proteolytic fragments produced by degradation of MHC in vivo. No proteolysis was detected in myofibrils during storage. These results help predict functional properties of fish proteins and changes during storage.     

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.     

Muscle protein changes post mortem in relation to pork quality traits

The relationship between post-mortem traits of muscle proteins and water loss traits was investigated using 84 pork loins representing the four quality traits of PSE, RSE (reddishpink, soft, exudative), RFN (reddish-pink, firm, non-exudative) and DFD. Protein solubility measurements (sarcoplasmic, myofibrillar and total) were lower and myosin denaturation (quantified by myofibrillar ATPase activity) was higher for PSE samples compared with samples from the other quality classes. RSE samples were similar to RFN samples in protein solubility and myosin denaturation, although RSE had lower values then DFD samples for protein solubility measurements. RFN samples had lower drip, thaw, cook and total water loss than RSE samples and all water loss traits were lowest for DFD samples and highest for PSE samples. Insoluble phosphorylase was the only characteristic that differentiated among PSE, RSE and RFN samples. SDS-PAGE and Western blots indicated that in PSE and RSE samples, the myofibrillar protein titin was less degraded and nebulin was more degraded compared with RFN and DFD samples. SDS-PAGE of extracted and unextracted myofibrils showed that the reduced myofibrillar solubility of PSE samples was caused by decreased extractability of the myosin heavy chain in these samples. In conclusion, although RSE samples have unacceptably high water loss, muscle protein denaturation was minimal and did not explain the low water-holding capacity.     

ABSENCE OF PROTEOLYSIS IN PURIFIED MYOFIBRILS OF POSTSPAWNED HAKE (MERLUCCIUS HUBBSI MARINI) DURING IN VITRO STORAGE AT 37C

The biochemical behavior of myofibrils from postspawned hake during in vitro storage at 37C was investigated. SDS‐PAGE, densitometric analysis of the band areas corresponding to the major myofibrillar proteins, and TCA soluble peptides determination showed no evidence of proteolysis in myofibrils after 44 h of incubation either in presence or in absence of a cocktail of protease inhibitors (1 mMPMSF+1mMiodoacetic acid + 1mM of EDTA). The absence of proteolysis in stored postspawned hake myofibrils contrasts with the autolysis in those from prespawned fish previously reported, indicating an influence of the reproductive cycle on the proteolytic activity closely associated to myofibrils.     

The storage life of iced southern blue whiting (Micromesistius australis)

The storage life of iced southern blue whiting (Micromesistius australis) was studied. Pre- and post-spawning characteristics were investigated by means of organoleptic assessments of raw and cooked fish, total volatile bases, reduced viscosity of high ionic strength muscle extract and GR Torrymeter readings. Results show a great influence of the biological condition on the keeping time in ice.
The results suggest that in pre-spawning condition the whole fish can be stored up to 12 days while in post-spawning condition the keeping time cannot exceed 6 days.
The gutting of the fish could lengthen the possible storage time in the post-spawning condition for up to 2 days. Nevertheless the post-spawning fish quickly loses its quality 2–3 days after catching.
The possibility of utilizing standard white fish processing machines and the influence of parasites are also discussed.     

Protein Denaturation in Pork Longissimus Muscle of Different Quality Groups

Differential scanning calorimetry (DSC) was used to investigate the protein denaturation characteristics of pork muscles from four quality groups namely RFN (red, firm, and non-exudative), RSE (red, soft, and exudative), PFN (pale, firm, and non-exudative), and PSE (pale, soft, and exudative). The thermograms indicated three endothermic peaks between 45°C to 90°C, corresponding to denaturation of myosin (peak I), sarcoplasmic proteins (peak II), and actin (peak III). The myosin peak was much reduced in PSE samples, while the actin peak remained almost identical in all groups. RFN and RSE samples were found to have very similar protein denaturation characteristics and were not significantly different in their thermodynamic protein denaturation parameters. PFN samples showed similar myofibrilar protein denaturation but significantly different sarcoplasmic protein denaturation characteristics compared to normal (RFN) samples according to their DSC thermograms. Based on these findings, it was suggested that the pale color in PFN pork is linked to sarcoplasmic protein denaturation.     

Freezing and frozen storage of actomyosin from different species

A comparative study was made of the influence of freezing (–24°C) and frozen storage (–12°C) on the functional properties (viscosity, solubility) and physico-chemical characteristics (aliphatic and aromatic hydrophobicity, ATPase activity) of actomyosin (AM) from myosystems (chicken and hake) of differing freezing and frozen stability. The difference in functional behaviour between chicken and hake AM as a consequence of freezing and frozen storage suggests that, for hake AM, denaturation and aggregation occur essentially through direct aggregation of AM molecules to produce AM aggregates, whereas in chicken proteins, AM first dissociates into myosin and actin to produce myosin and myosin-actin aggregates.     

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     

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.     

不同冷冻方式对调味鱼贮藏期间肌原纤维蛋白的影响

为探究不同冷冻方式对调味鱼冻藏品质的影响,提取快速（－80 ℃）、中速（－30 ℃）、慢速 （－18 ℃）冷冻调味鱼贮藏期间的肌原纤维蛋白,测定相关蛋白变性指标,并用电子鼻对气味进行测定。结果 表明：随着贮藏时间的延长,3 种冷冻处理调味鱼的肌原纤维蛋白均发生不同程度的氧化,具体表现为羰基含 量、表面疏水性不断上升,活性巯基含量、乳化性及稳定性不断下降;－80 ℃冷冻调味鱼的肌原纤维蛋白变性 程度最低,－18 ℃冷冻调味鱼的肌原纤维蛋白变性最严重;－30、－80 ℃冷冻调味鱼贮藏期间气味变化较小, 而－18 ℃冷冻调味鱼贮藏期间气味变化较大。     

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).     

Influence of Gonadal Stage of Hake (Merluccius Hubbsi Marini) on Biochemical Properties of Myofibrils Stored at 2 to 4 °C

ABSTRACT: The influence of the gonadal stage of hake on the biochemical properties of myofibrils stored at 2 to 4 °C was studied. At 0 time and during storage, both Mg²⁺-Ca²⁺-ATPase activity and Ca²⁺ sensitivity of myofibrils from post-spawned hake were significantly higher (p < 0.01) than those of pre-spawned fish. The profiles of SDS-PAGE gels of unstored and stored myofibrils from pre-spawned hake showed a partially denatured myosin heavy chain. The actin-myosin ratio in myofibrils from pre-spawned hake was significantly lower (p < 0.01) than the ratio in post-spawned hake. Irrespective of the gonadal condition of fish, no changes in the myosin-actin ratio of stored myofibrils were observed.