Thermal Aggregation and Dynamic Rheological Properties of Pacific Whiting and Cod Myosins as Affected by Heating Rate

Thermal aggregation of cod myosin and Pacific whiting myosin were compared at different heating rates. Turbidity of Pacific whiting started to increase at 30°C and decreased at >50°C, and fluorescence intensity of ANS-myosin of both species was greater at slower heating rates at 20–80°C. Storage modulus (G′) of cod myosin gradually increased as temperature increased resulting in more elastic gel at slower heating rates. G′ of whiting myosin rapidly increased at 30°C, maximized at 45–48°C and decreased afterward. The onset temperature of a decreased G′ shifted from 45°C to 49°C as heating rate increased from 0.5 to 2°C/min. Whiting myosin heated at 2°C/min retained more myosin heavy chain.     

Influence of Heating and Cooling Rates on Bacillus cereus Spore Survival and Growth in a Broth Medium and in Rice

Survival, spore germination, and growth of emetic and diarrheal type strains of Bacillus cereus were evaluated in broth and rice media during heating and cooling. Samples were heated to 80°C (20C°/hr or 40C°/hr) or 90°C (ca. 900C°/hr), prior to cooling to 10°C (5C°/hr or 10C°/hr). Following heating to 80°C, growth occurred during 5C°/hr cooling. After heating to 90°C, inactivation of three strains occurred during cooling from 90 to 80°C and again from 50 to 40°C. Great variability was observed among the responses of the four strains. Emetic strains exhibited greater survival than diarrheal strains. Rice reduced low temperature inactivation, and did not favor emetic strains. Significant two and three way interactions existed among media, strains, heating and cooling rates.     

Rapid Heating of Alaska Pollock and Chicken Breast Myofibrillar Proteins as Affecting Gel Rheological Properties

Surimi seafoods (fish/poikilotherm protein) in the U.S.A. are typically cooked rapidly to 90+°C, while comminuted products made from land animals (meat/homeotherm protein) are purposely cooked much more slowly, and to lower endpoint temperatures (near 70 °C). We studied heating rate (0.5, 25, or 90 °C/min) and endpoint temperature (45 to 90 °C) effects on rheological properties (fracture, small strain) of washed myofibril gels derived from fish (Alaska pollock) compared with chicken breast at a common pH (6.75). This was contrasted with published data on gelation kinetics of chicken myosin over the same temperature range. Heating rate had no effect on fracture properties of fish gels but slow heating did yield somewhat stronger, but not more deformable, chicken gels. Maximum gel strength by rapid heating could be achieved within 5 min holding after less than 1 min heating time. Dynamic testing by small strain revealed poor correspondence of the present data to that published for gelling response of chicken breast myosin in the same temperature range. The common practice of reporting small‐strain rheological parameters measured at the endpoint temperature was also shown to be misleading, since upon cooling, there was much less difference in rigidity between rapidly and slowly heated gels for either species.     

Thermal modifications of structure and codenaturation of α‐lactalbumin and β‐lactoglobulin induce changes of solubility and susceptibility to proteases

Study of heat denaturation of major whey proteins (β‐lactoglobulin or α‐lactalbumin) either in separated purified forms, or in forms present in fresh industrial whey or in recomposed mixture respecting whey proportions, indicated significant differences in their denaturation depending on pH, temperature of heating, presence or absence of other co‐denaturation partner, and of existence of a previous thermal pretreatment (industrial whey). α‐Lactalbumin, usually resistant to tryptic hydrolysis, aggregated after heating at ⪈85°C. After its denaturation, α‐lactalbumin was susceptible to tryptic hydrolysis probably because of exposure of its previously hidden tryptic cleavage sites (Lys‐X and Arg‐X bonds). Heating over 85°C of β‐lactoglobulin increased its aggregation and exposure of its peptic cleavage sites. The co‐denaturation of α‐lactalbumin with β‐lactoglobulin increased their aggregation and resulted in complete exposure of β‐lactoglobulin peptic cleavage sites and partial unveiling of α‐lactalbumin tryptic cleavage sites. The exposure of α‐lactalbumin tryptic cleavage sites was slightly enhanced when the α‐lactalbumin/β‐lactoglobulin mixture was heated at pH 7.5. Co‐denaturation of fresh whey by heating at 95°C and pH 4.5 and above produced aggregates stabilized mostly by covalent disulfide bonds easily reduced by β‐mercaptoethanol. The aggregates stabilized by covalent bonds other than disulfide arose from a same thermal treatment but performed at pH 3.5. Thermal treatment of whey at pH 7.5 considerably enhanced tryptic and peptic hydrolysis of both major proteins.     

Ohmic Heating Maximizes Gel Functionality of Pacific Whiting Surimi

Surimi without enzyme inhibitors containing 78% moisture and 2% NaCl was heated conventionally and ohmically to 90°C after holding at 55°C for 0, 1, 3 and 5 min. Gels heated slowly in a water bath exhibited poor gel quality, while the ohmically heated gels without holding at 55°C showed more than a twofold increase in shear stress and shear strain over conventionally heated gels. Degradation of myosin and actin was minimized by ohmic heating, resulting in a continuous network structure. Ohmic heating with a rapid heating rate was an effective method for maximizing gel functionality of Pacific whiting surimi without enzyme inhibitors.     

Comparative study of the nutritive value of casein heated by microwave and conventionally

The objective of this study was to compare the net protein utilisation, digestibility and biological value of casein solution heated by microwave or conventionally, ie on an electrically heated plate. Portions (500 ml) of an aqueous solution of casein (100 g litre^?1) were heated to a temperature of 80°C and subsequently kept at 80± 5°C for 2 min. The heating times of microwave and conventional heating were the same. Additional portions were subjected to a so-called abused treatment consisting of repeating the normal microwave or conventional heat treatment three times. Next, the nutritive value of heated and unheated casein was determined in a 10-day feeding study with Wistar rats. The net protein utilisation, digestibility and biological value of unheated casein were 61.1, 100 and 61%, respectively. Comparable values were obtained for casein heated by microwave or conventionally. These results did not therefore indicate any decrease in nutritive quality of the protein by microwave heating or conventional heating under simulated household conditions.     

Effect of thermochemical treatment on the structure of inulin and its gelling properties

The aim of this study was to investigate rheological properties of inulin gels and the changes in the inulin structure after heating the high polymerised inulin (DP ≥ 23) solutions at different pH. The 20% inulin solutions were heated at pH 3, 5 and 7 at 60 and 80 °C followed by 21 h storage at 5 and 25 °C. Rheological properties evaluation indicated that the increase in the heating time and temperature significantly (P ≤ 0.05) influenced on the inulin structure. Reducing sugars, cryoscopy and HPLC analysis revealed that chemical structure of inulin is stable in neutral and slightly acidic conditions (pH 5). Gelation of the high polymerised inulin solution after heating at pH 7 and 80 °C could be inhibited after dissolving the inulin crystallites which act like seeding crystals. At lower temperatures in which, not all crystallites were dissolved; it was feasible for the solutions to form firm inulin gels.     

Effect of heating temperature and duration on the texture and protein composition of Bighead Carp (Aristichthys nobilis) muscle

The effect of heating temperature and duration on the texture and protein composition of bighead carp (Aristichthys nobilis) muscle was investigated. Samples were heated at 90°C, 100°C, 110°C, and 120°C for different times. The results showed that cooking loss increased significantly with increasing temperature and heating time, and a fractional conversion model was used to describe the increase. Hardness, chewiness, and gumminess exhibited a similar four-phase change with two peaks at 90°C, 100°C, and 110°C, and the trend was not observed at 120°C. Analysis of sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) revealed that contents of myosin heavy chain decreased entirely to disappear and contents of actin significantly decreased. Increasing the intensity of the heavy molecular weight bands indicated the aggregation of myosin. Content of water-soluble proteins presented an increasing trend, which reflected the protein degradation and the release of low-molecular-weight compounds.     

The mechanism of formation of gels from myosin molecules

Heat-set myosin gels form the basis of the adhesive that binds particles of meat together in meat products. The manner in which a gel network is formed from myosin has been investigated by studying the aggregates produced when dilute solutions of rabbit skeletal myosin molecules in 0.6 M KCI, 20 mM potassium phosphate, pH 6.5, were heated at a single temperature between 30 and 60°C. The aggregates have been examined by transmission electron microscopy after either negative staining or rotary shadowing. After heating at 30°C for 30 min. no change in structure is detected. After heating at 35°C for 30 min, the two heads of some myosin molecules coalesce and some dimers are formed by aggregation through the heads. After heating at 40°C for 30 min. up to about 13 myosin molecules aggregate through their heads to form a globular mass up to 60 nm across with the tails radiating outwards. At higher temperatures such head-linked oligomers aggregate further. At 48°C oligomers coexist with aggregates formed by the coalescence of two or more oligomers. In such aggregates the globular masses are in close proximity and tails radiate from them. After heating myosin at 50°C, aggregates are formed by the coalescence of more oligomers and the tails are seen only indistinctly. At 60°C the particles formed contain a large number of globular masses and are typically 100 to 200 nm across, occasionally up to 1 μm. It is possible that the globules making up the strands of the gel networks in scanning electron micrographs are composed of similar particles. It is suggested that these globules are formed by head-head interactions but that tail-tail interactions may be important in forming the strands and cross-links of the gel network. When a heat-set myosin gel is compacted by centrifugation, the supernate contains essentially all the LC1 and LC3 light chains of the parent molecule but only a small fraction of the LC2 light chains. We suppose that dissociation of the LC1 and LC3 chains from the heads creates hydrophobic patches which cause intramolecular and intermolecular head association.     

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.     

Impacts of yeast β-glucan on thermal aggregation and flavour adsorption capacity of Spanish mackerel myosin

The effects of yeast β-glucan (YG) on the aggregation and flavour adsorption capacity of Spanish mackerel myosin under two-step heat treatments were investigated. The surface hydrophobicity and active sulfhydryl content of heat-induced myosin was increased by the addition of YG, reaching the maximum values of 81.46 and 17.49 mol per 10⁵ g after heating at 90 °C for 10 min. YG incorporation significantly decreased the α-helix content of myosin to the minimum value of 48.27%, whereas the β-sheet was increased. The addition of YG promoted the unfolding of myosin, exposing more hydrophobic bonding sites, thus enhancing the flavour-binding capacity of myosin. Lengthy heating at 90 °C combined with YG addition accelerated the cross-linking and aggregation of myosin, leading to the re-embedding of the exposed hydrophobic groups as much as the reduction of flavour-binding sites. However, YG's unique porous structure could also boost the flavour adsorption ability of myosin containing YG.     

Rheological and microstructural characterization of double cream cheese

Firmness at 5 °C, dynamic viscoelastic moduli (G' G) and capillary extrusion profiles at 20 °C, were obtained on double cream cheese after the different steps of processing (curd obtention, mixing at 70 °C and 500 r.p.m. with heat-denatured WPC, heating at 85 °C, homogenizing at 20 MPa (1st stage) and 5 MPa (2nd stage) and cooling to 20 °C and 13 °C) and storage 7–9 days at 5 °C. It was found that double cream cheese became firmer and more elastic after heating and homogenization, although it became softer and more viscous after mixing and cooling. Transmission Electron Microscopy (TEM) and cryo-Scanning Electron Microscopy (SEM) micrographs showed that rheology results could be related to aggregation (during heating and homogenization) and disruption (during cooling) of milk fat globule/casein complexes. Dispersion of homogenization clusters after cooling, and aggregation of milk fat globules during storage caused double cream cheese structural instability to appear. It was suggested that the heterogeneity of capillary extrusion profiles could be quantified through application of fractal concepts and Fourier analysis and related to structure and texture of double cream cheese.     

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).     

Heat-induced Gelation of Chicken Gizzard Myosin

Chicken gizzard myosin solution formed a gel when heated above 40°C. The rigidity of the gel was constant above 65°C. Maximum pH for gel formation was 5.9 at 0.6M and 5.7 at 0.15M KCl. Higher rigidity of the myosin gel was observed at low ionic strength than at high ionic strength. Rigidities of myosin at 0.6M KCl increased by (mg/mL)^2.5 and at 0.15M (mg/mL)^{1, 4} myosin concentration. The strength of gizzard myosin gels was comparable to that of myosin gels from chicken breast muscle under similar conditions.     

Heating of β-Lactoglobulin A Solution in a Closed System at High Temperatures

ABSTRACT β-Lactoglobulin A solution at pH 6.4 was heated to 180 °C at the rate of 6 °C/min. By differential scanning calorimetry two independent endothermic peaks were observed. The first peak appeared below 100 °C is corresponded to the thermal denaturation of protein. This conformational change led to the aggregation and polymerization of molecules through disulfide linkage, particularly observed above 100 °C. The second endothermic peak appeared around 150 °C, which was brought by the decomposition of molecules as judged from electrophoresis. Up to 100 °C the viscosity of β-lactoglobulin A solution increased by heating, while the viscosity was reduced beyond 113 °C, due to change in the size of aggregate and decomposition of β-lactoglobulin A molecules.     

Disulfide-mediated Polymerization Reactions and Physical Properties of Heated WPI-stabilized Emulsions

Heating a 19 wt% corn oil-in-water emulsion stabilized by 1 wt% whey protein isolate from 30 to 70°C and then cooling to 25°C for at least 15 hr, brought about minimal changes in droplet aggregation, apparent viscosity and susceptibility to creaming. At 75°C, droplet aggregation occurred but this decreased on heating to 90°C. The apparent viscosity and susceptibility of droplets to creaming increased as the degree of droplet aggregation increased. Inclusion of the sulfhydryl blocking agent N-ethylmaleimide to inhibit thiol/disulfide interchange reactions did not affect droplet aggregation but resulted in higher apparent viscosity values and susceptibility to creaming at 85 and 90°C and not at lower temperatures. The results suggest that droplet aggregation results from noncovalent interactions between unfolded protein molecules adsorbed on different droplets and that the interactions are strengthened by disulfide bonds.     

Protein-Protein Interaction of Fish Myosin Fragments

Protein-protein interaction of myosin fragments from flying fish and white marlin muscles was studied by means of absorbance changes resulting from aggregation at temperatures of 20°C to 70°C. Subfragment-1 (S-1) exhibited a high extent of interaction with the transition temperature of 35–36°C, while the interaction of heavy meromyosin (HMM) was very weak. Though light meromyosin (LMM) gave lower interaction values throughout the heating temperature, the addition of butanol promoted markedly the interactions at the temperature above 50°C. The degree of promotion was high for flying fish and low for white marlin.     

Some Effects on the Mechanical Properties of Meat Produced by Cooking at Temperatures between 50° and 60°C

Peak force (PF) shear values obtained for stretched muscles from beef animals of three ages (2–3 months, 2–6 year and 12–17 years) decreased when heated for 1 hr at temperatures above 50°, 55° and 60°C respectively. PF values obtained for veal muscles were unaffected by heating at 50°C for up to 8 hr but rapidly decreased with increased time of heating at 55° or 60°C. The decrease in shear values with heating time was still evident after the connective tissue contribution was eliminated by a further cook at 80°C. Samples from the oldest animals required 24–48 hr at 60°C to produce a large decrease in the connective tissue contribution. Tenderization by prolonged cooking at 50–60°C was achieved by accelerated aging of the myofibrillar structure and, at ≥ 55°C, by a weakening of the collagenous connective tissue also.     

Herring Surimi During Low Temperature Setting, Physicochemical and Textural Properties

Herring surimi was held at 10°C for 0 to 24 hr, then heated at 90°C for 30 min. Rigidity, shear stress and shear strain at failure increased with holding time at 10°C, and the values of shear strain were 2.0 when setting time was > 3 hr. Electrophoresis revealed three types of protein-protein interactions involved in the formation of myosin polymers during setting at 10°C and/or subsequent heating at 90°C. The interactions included covalent, nondisulfide interactions; disulfide bonds and noncovalent interactions. The contributions of each type of protein-protein interaction to the formation of myosin polymers varied with setting time. Variations in the types of interactions were related to physical/chemical properties.     

Carp Natural Actomyosin: Thermal Denaturation Mechanism

Structural changes of actomyosin, the major protein of muscle, on heating have been estimated on ATPase activity. We investigated carp actomyosin molecule changes on heating based on biophysical and biochemical techniques. Actomyosin molecules began to unfold at ～30°C. Hydrophobic amino acid residues and SH groups, which had been inside the molecule, emerged to the surface. Because of hydrophobic interactions and disulfide bonds, actomyosin molecules formed aggregates. At > 40°C, a part of myosin molecules was dissociated from actin filaments. Thus, dissociated myosin and the myosin-lacking molecules co-existed. In addition, fragmentation of actin filaments was observed, which was associated with the dissociation of myosin molecules. At ≥ 60 °C actomyosin molecules formed larger aggregates, in which no filamentous shape was observed. This aggregation occurred mainly by formation of SS bonds.