Dynamic rheological measurements on heat-induced myosin gels: Effect of ionic strength,protein concentration and addition of adenosine triphosphate or pyrophosphate

Myosin solutions and suspensions have been monitored during heating at pH 6.0 by using dynamic rheological measurements. The storage modulus (G′), the loss modulus (G) and the phase angle (δ) all showed a marked dependence on ionic strength in the temperature range 25–75°C. The filamentous gels (ionic strength <0.34) displayed a temporary reduction in G′ at temperatures between 50 and 60°C, presumably due to denaturation in parts of the rod portion of the myosin molecule. In the same temperature region the concentration dependence of G′ changed by a power of 2. The loss modulus also showed a marked concentration dependence, while the phase angle varied with concentration primarily at low (<50°C) temperatures. For the final gels, heated to 75°C, only G′ indicated marked differences due to different protein concentrations and ionic strengths; all gels were almost completely elastic (δ?1°). Adenosine triphosphate was shown to have a pronounced temporary effect on the filamentous gel formed at low temperatures, i.e. on the gel with the highest concentration dependence, while pyrophosphate had no such effect. However, both adenosine triphosphate (or rather its hydrolysis product: adenosine diphosphate) and pyrophosphate appeared to have a small, lasting effect on the heat-gelling ability of myosin: the former a detrimental effect, the latter an improvement.     

Mobility and redistribution of waters within bighead carp (Aristichthys nobilis) heat-induced myosin gels

Water-holding capacity is closely related to gel microstructure, and is a very important quality trait in surimi and surimi product. The changes in the secondary structure, gel microstructure, and the migration of water in bighead carp (Aristichthys nobilis) myosin gel induced by different temperatures (50–90°C) were investigated. The α-helical structure of myosin decreased at temperatures of 40°C or higher. The fractal dimension of the gels increased at 40, 50, and 60°C, but decreased at temperatures over 60°C. The pore size of the gels increased with temperatures up to 50°C, decreased at 60°C, and then increased with temperatures up to 90°C again. The transverse relaxation times also varied; T₂₁ remained constant at temperatures over 40°C; T₂₂ decreased at temperatures lower than 50°C, increased at 60°C, and then decreased with temperatures up to 90°C; and T₂₃ increased at temperatures lower than 50°C and then remained constant until 90°C. Principal component analysis showed that the proportion of T₂₂ water (PT₂₂) was inversely correlated with the unfolding of myosin, whereas directly correlated with the pore size. The proportion of T₂₃ water (PT₂₃) was positively correlated with the fractal dimensions of the gels, whereas negatively correlated with the pore size. The migration of the secondary layer of water was mainly caused by hydrophobic force and the physical space formed by the myosin backbone, and the migration of water within the third layer was mainly caused by capillary pressure. Therefore, the mobility and redistribution of waters depend on the water retention mechanism, which is determined by the physical structure of gels. This study provides further information about the relationship between the NMR data, gel microstructure, water mobility, and distribution.     

Transglutaminase treatment of pea proteins: Effect on physicochemical and rheological properties of heat-induced protein gels

Rheological properties of heat-induced pea protein isolate (PPI) gels with added microbial transglutaminase (MTGase) were studied under various reaction conditions. A positive linear relationship was observed between level of MTGase used (0 to 0.7% w/w) and shear stress and shear strain of heat-set commercial pea protein isolate (PPIc) gels at 92 °C following incubation at 50 °C. Use of MTGase allowed for preparation of PPIc gels of similar strength and elasticity as commercial soy protein isolate gels and commercial meat bologna. MTGase treatment did not alter thermal properties of PPI gels. The shear stress and strain of PPIc gels were also improved following low temperature (4 °C) incubation of PPI with MTGase. Enhancement of shear strain or gel elasticity of heat-induced PPI gels with MTGase has not been reported before and provides opportunities for extending the properties of pea proteins when developing new food products.     

Dynamic rheological and thermal study of the heat-induced gelation of tomato-seed proteins

The structural aspects of proteins prepared from tomato seeds were studied by dynamic rheology, modulated differential scanning calorimetry (MDSC) and Fourier transform infrared (FTIR) spectroscopy. The critical gel point was determined from storage (G’) and loss (G”) moduli as functions of time and frequency. The findings indicate that tomato-seed proteins can create gels during heat treatment at 90–95 °C. Mechanical spectra of the tomato-seed protein gels were classified as weak gels based on the frequency sweep, complex viscosity (η∗) and tan δ results. Moreover, G’ and G” changes were found to be dependent on both concentration and frequency. MDSC showed two peaks, with gelation temperatures (T_d) of 70 and 87 °C, which we attribute to starch and globular protein fractions, respectively. The FTIR spectra show an increase in the absorbance of amides I and II after increasing the solid content from 1 to 10% w/w, indicating mostly β-sheet and α-helical conformations.     

Native vs. damaged milk fat globules: membrane properties affect the viscoelasticity of milk gels

The storage modulus G' of rennet and acid milk gels filled with milk fat globules was measured as a function of the fat globule surface composition (native milk fat globule membrane, caseins and whey proteins, or a mixture of the three due to mechanical treatments) and surface area (i.e., the fat globule size). By different technological procedures, it was possible to obtain fat globules of constant surface composition but various sizes, and vice-versa, which had never been done. For both rennet and acid gels, a critical fraction of the fat globule surface covered by caseins and whey proteins was identified (approximately 40%), beyond which G' increased. Below this threshold, the gel viscoelasticity was unaffected by mechanical treatments. When the diameter of native milk fat globules decreased, the G' of rennet gels increased slightly, whereas that of acid gels decreased sharply. For both types of gels, G' increased when the diameter of partially disrupted fat globules decreased. For recombined globules completely covered with caseins and few whey proteins, G' increased with fat globule surface area for rennet gels whereas it decreased for acid gels. With the help of confocal microscopy and in the light of general structural differences between rennet and acid gels, a mechanism is proposed for the effect of fat globule damage and diameter on G', depending on the interactions the globules can undergo with the casein network.     

Interactions between soy protein isolate and xanthan in heat-induced gels: The effect of salt addition

The influence of xanthan and/or KCl addition on the properties of heat-induced soy protein isolate (SPI) gels at pH 3.0 was studied. Changes in protein solubility and subunit composition as well as in the mechanical properties, microstructure and water holding capacity of the gels were determined. The effect of KCl addition on each biopolymer solution was also investigated. The results indicated that SPI–xanthan gels prepared without KCl were mainly stabilized by non-covalent (H-bonding and hydrophobic) and SS bond interactions, whereas in gels containing KCl, electrostatic interactions were also involved in maintaining the gel structure. The β-7S subunit was probably the fraction electrostatically linked to the xanthan. The different values found for the mechanical properties after the addition of xanthan and/or KCl, were associated with the coarseness of the gel. Xanthan and KCl probably showed a synergistic effect on the Young modulus because KCl induced a conformation transition of the xanthan molecules.     

Effect of controlled kappa-casein hydrolysis on rheological properties of acid milk gels

An experimental method based on the controlled chymosin-induced kappa-casein hydrolysis of milk was proposed to modify micellar reactivity. Milk samples with a degree of kappa-casein hydrolysis of 19, 35, and 51% were obtained. The physicochemical properties of partially converted casein micelles were determined. The net negative charge of casein micelles was reduced with increasing degree of kappa-casein hydrolysis and a small but significant decrease in hydrodynamic diameter and micellar hydration were noted. Dynamic low amplitude oscillatory rheology was used to monitor the rheological properties of acid milk gels (GDL) made with partially chymosin-hydrolyzed milks in comparison with those of strictly acid and rennet gels. An increase in the gelation pH value was observed with increasing the degree of kappa-casein hydrolysis. The moduli values (G' and G') reached 2 h after the point of gel were, for all degrees of hydrolysis tested, significantly higher than those of strictly rennet and acid gels. Comparison of changes in delta G'/delta t with time indicated differences in gel formation that could be related to the increased values of G' obtained for acid gel made with chymosin-treated milk. At a given time after gelation (2 h), increasing the degree of kappa-casein hydrolysis in milk led also to an increase in the loss tangent and the serum holding capacity of acid milk gels suggesting a correlation between these two parameters.     

Molecular forces involved in heat-induced pea protein gelation: Effects of various reagents on the rheological properties of salt-extracted pea protein gels

The molecular forces involved in the gelation of heat-induced pea protein gel were studied by monitoring changes in gelation properties in the presence of different chemicals. At 0.3 M concentration, sodium thiocyanate (NaSCN) and sodium chloride (NaCl) showed more chaotropic characteristic and enhanced the gel stiffness, whereas sodium sulfate (Na₂SO₄) and sodium acetate (CH₃COONa) stabilized protein structure as noted by increasing denaturation temperatures (T_d) resulting in reduced storage moduli (G′). To determine the involvement of non-covalent bonds in pea protein gelation, guanidine hydrochloride (GuHCl), propylene glycol (PG), and urea were employed. The significant decrease in G′ of pea protein gels with the addition of 3 M GuHCl and 5 M urea indicated that hydrophobic interactions and hydrogen bonds are probably involved in pea protein gel formation. The increase in G′ with increasing PG concentration (5–20%), demonstrated hydrogen bonds and electrostatic interaction involvement. No significant influence was observed on G′ with addition of different concentrations of β-mercaptoethanol (2-ME), low levels of dithiothreitol (DTT), and up to 25 mM N-ethylmaleimide (NEM), which indicated that disulfide bonds are not required for gel formation, but data at higher DTT and NEM concentrations and slow cooling rates showed a minor contribution by disulfide bonds. Reheating and recooling demonstrated that gel strengthening during the cooling phase was thermally reversible but not all the hydrogen bonds disrupted in the reheating stage were recovered when recooled.     

Influence of partially demineralized milk proteins on rheological properties and microstructure of acid gels

Innovative clean label processes employed in the manufacture of acid gels are targeted to modify the structure of proteins that contribute to rheological properties. In the present study, CO₂-treated milk protein concentrate powder with 80% protein in dry matter (TMPC80) was mixed with nonfat dry milk (NDM) in different ratios for the manufacture of acid gels. Dispersions of NDM and TMPC80 that provided 100, 90, 70, and 40% of protein from NDM were reconstituted to 4.0% (wt/wt) protein and 12.0% (wt/wt) total solids. Dispersions were adjusted to pH 6.5, followed by heat treatment at 90°C for 10 min. Glucono-δ-lactone was added and samples were incubated at 30°C, reaching pH 4.5 ± 0.05 after 4 h of incubation. Glucono-δ-lactone levels were adjusted to compensate for the lower buffering capacity of samples with higher proportions of TMPC80, which is attributable to the depletion of buffering minerals from both the serum and micellar phase during preparation of TMPC80. Sodium dodecyl sulfate-PAGE analysis indicated a higher amount of caseins in the supernatant of unheated suspensions with increasing proportions of CO₂-treated TMPC80, attributable to the partial disruption of casein micelles in TMPC80. Heat treatment reduced the level of whey proteins in the supernatant due to the heat-induced association of whey proteins with casein micelles, the extent of which was larger in samples containing more micellar casein (i.e., samples with a lower proportion of TMPC80). Particle size analysis showed only small differences between nonheated and heated dispersions. Gelation pH increased from ?5.1 to ?5.3, and the storage modulus of the gels at pH 4.5 increased from ?300 to ?420 Pa when the proportion of protein contributed by TMPC80 increased from 0 to 60%. Water-holding capacity also increased and gel porosity decreased with increasing proportion of protein contributed by TMPC80. The observed gel properties were in line with microstructural observations by confocal microscopy, wherein sample gels containing increasing levels of TMPC80 exhibited smaller, well-connected aggregates with uniform, homogeneous pore sizes. We concluded that TMPC80 can be used to partially replace NDM as a protein source to improve rheological and water-holding properties in acid gels. The resultant gels also exhibited decreased buffering, which can improve the productive capacity of yogurt manufacturing plants. Overall, the process can be leveraged to reduce the amount of hydrocolloids added to improve yogurt consistency and water-holding capacity, thus providing a path to meet consumer expectations of clean label products.     

Dynamic rheological behaviour and biochemical properties of rabbit skeletal actomyosin during storage at 0° C

The relationship between the dynamic rheological properties of heat induced gels of actomyosin (natural actomyosin) and the denaturation of actin in actomyosin during storage without ATP at pH 6.0 and 0°C was investigated using biochemical and dynamic rheological measurements. The complex modulus of gels after heating actomyosin containing 0.5 or 1.5 M KCl (pH 6.0) at 80°C increased with increasing storage time. The dynamic rheological behaviour during heat gelation of actomyosin in 1.5 M KCl indicated that the first rheological transition peak in the 50–53°C range induced by the presence of F-actin gradually disappeared with increasing storage time. However, in 0.5 M KCl, this transition peak clearly remained even after 15 days. The time course of denaturation of actin in actomyosin treated with 1.5 M KCl at pH 6.0 showed an increase in the percent denaturation after the storage was started, and about 100% of the actin became denatured after 21 days. In the case of 0.5 M KCl, unlike 1.5 M, the denaturation of actin occurred quickly within the first 5 days and then did not proceed. A sigmoidal relationship was found between the percent denaturation of actin and the KCl concentration added, the greatest change occurring at KCl concentrations between 0.5 and 1.0 M. The data indicated that the change in the property of actin in actomyosin during storage at low temperature exerts a great influence on the viscoelasticity of heat-induced gels of actomyosin.     

On the formulation design and rheological evaluations of pectin-based functional gels

In the current study, formulation design and development of a novel pectin-based functional gel were investigated. Amidated low methoxyl pectin (ALMP), high methoxyl pectin (HMP), sorbitol, inulin, rebaudioside-A, gardenia, phloridzin, quercetin, apple flavor, and calcium chloride (as its dihydrated salt) were selected as general ingredients. Response surface methodology was applied to design different formulations and to investigate on their experimental responses. The oscillatory tests were carried out in 2 stages with ALMP, HMP, sorbitol, and inulin as the variables of the 1st stage and ALMP and calcium chloride as the variables of the 2nd stage. Results of the 1st stage indicated relatively similar behaviors throughout the frequency range applied for all of the samples studied (true gels). However, magnitudes of the 5 rheological parameters of this study (storage modulus, loss modulus, loss tangent, complex modulus, and complex viscosity) were affected by different variables (ALMP, HMP, inulin, and sorbitol concentrations). Experimental results confirmed that sorbitol could be omitted from the formula while inulin and HMP could be used at their highest levels studied. At the 2nd stage, only 2 formulas indicated a strong gel behavior and other formulas showed typical behaviors of weaker gels or those of the concentrated solutions. At the conclusion of this study, a finished gelled product formula was suggested with the application of the best levels of the ingredients. The resultant gel was found to be set rapidly with no syneresis and showed a potential to be considered as a functional gelled dessert.     

The suitability of muscle of Cirrhinus mrigala in the formation of gel: a comparative electrophoretic study of six tropical carp meats

The annual inland fish production in India is 3.2 million tonnes and above. Aquaculture is around 80% of total inland fish production. The enhanced aquaculture production demands alternate processing methods for better utilisation of the farmed fish. Cirrhinus mrigala is one of the dominated species among cultured Indian major carps, but rated with lowest price. Fish meat with good gel‐forming capacity is a prerequisite in the production of fast‐moving fabricated analogue product. Cirrhinus mrigala had the highest gel strength, i.e. 435 gcm of its fresh meat in comparison with other carps. Apart from the highest protein and salt‐soluble protein nitrogen content in fresh C. mrigala meat, this article also reports that the presence of the darkest thick band of myosin heavy chain and actin in the polyacrylamide gel electrophoresis pattern of salt‐soluble extract of C. mrigala might be another possible reason for the highest gel strength in comparison with other carps. Cirrhinus mrigala contained white meat around 90%. This carp could be suitably used in the preparation of high‐valued fabricated analogue product.     

Effect of oxidoreduction potential and of gas bubbling on rheological properties and microstructure of acid skim milk gels acidified with glucono-δ-lactone

Milk oxidoreduction potential was modified using gases during the production of a model dairy product and its effect on gel setting was studied. Acidification by glucono-δ-lactone was used to examine the physicochemistry of gelation and to avoid variations due to microorganisms sensitive to oxidoreduction potential. Four conditions of oxidoreduction potential were applied to milk: milk was gassed with air, nongassed, gassed with N₂, or gassed with N₂H₂. The rheological properties and microstructure of these gels were determined using viscoelasticimetry, measurement of whey separation, and confocal laser scanning microscopy. It appeared that a reducing environment led to less-aggregated proteins within the matrix and consequently decreased whey separation significantly. The use of gas to modify oxidoreduction potential is a possible way to improve the quality of dairy products.     

Study on the rheological properties of heat-induced whey protein isolate-dextran conjugate gel

Effect of glycosylation on the rheological properties of whey protein isolate (WPI) during the heat-induced gelation process was evaluated. Significant changes in browning intensity, free amino groups content and SDS-PAGE profile showed that the conjugate of WPI and dextran (150 kDa) was successfully prepared using the traditional dry-heating treatment. For the conjugate, during the heating and cooling cycle, the curves of G′ and G″ were considerably shifted to lower values and their shapes varied comparing to the corresponding spectra of initial WPI and WPI + dextran mixture. After holding at 25 °C, G' reached a value of about 2200 Pa, only a tenth of the value that obtained in the initial WPI gel. Moreover, frequency sweep measurements revealed that the stiffness of gel was greatly reduced in the conjugate, although a typical elastic gel was still formed. All data showed that the rheological properties of thermal gelation could be modified upon the covalent attachment of dextran.     

Effect of preheating temperature on the microstructure of walleye pollack surimi gels under the inhibition of the polymerisation and degradation of myosin heavy chain

BACKGROUND: The physical attribute of heat‐induced gel texture is highly dependent on the microstructure of the gel. In this study the microstructures of walleye pollack surimi gels preheated at various temperatures with and without inhibitors (ethylenediamine‐N,N,N′,N′‐tetraacetic acid, iodoacetamide and leupeptin) were observed with a natural scanning electron microscope. RESULTS: Without inhibitors, gels preheated at 30 °C showed a fine and uniform network structure together with the highest polymerisation of myosin heavy chain (MHC) and the highest gel strength. At 60 °C, gels exhibited a broken, disrupted and loose cluster‐like structure together with the highest degradation of MHC and the lowest gel strength. Under the inhibition of polymerisation and degradation of MHC a fine network was observed up to 40 °C during preheating. However, after a second step of heating at 80 °C the microstructures were disrupted and resembled each other regardless of the preheating temperature. CONCLUSION: Heat‐induced gel formation is related to the polymerisation and degradation of MHC and the microstructure of the gel during preheating. Gelation occurred during setting even under the inhibitory condition, and the formation of covalent bonding by transglutaminase is not essential to the formation of a three‐dimensional network during setting but is essential to the gel strength enhancement effect of setting by subsequent heating at 80 °C. Copyright © 2010 Society of Chemical Industry     

Influence of fibre addition on the rheological properties of peach juice

The addition of dietary fibres is a tendency in several foods. Understanding the changes in food sensory and physical properties due to fibre addition is thus, essential for food process design. The present work has evaluated the influence of peach fibre addition on the rheological properties of peach juice. Flow behaviour and influence of fibre concentration on Herschel‐Bulkley model’s parameters were evaluated. The flow behaviour of the products was changed due to fibre addition, from Newtonian to pseudoplastic and then, Herschel‐Bulkley behaviour. The parameters could be well modelled by exponential (σ₀), power law (k) and sigmoidal (n) functions (R² > 0.98). The viscoelastic properties were evaluated for the most concentrated products. Variation of storage and loss modulus with the oscillatory frequency were well described by a power function (R² > 0.96), and dependency of its parameters with temperature was well modelled by Arrhenius’ model (k′, k″, R² > 0.94) and quadratic function (n′, n″, R² > 0.90).     

大豆11S球蛋白凝胶形成的影响因素分析

以低温脱脂大豆粕为原料,采用等电点冷沉法浸提11S球蛋白。对11S球蛋白凝胶形成过程中影响凝胶质构的因素进行了分析,研究表明,11S球蛋白凝胶形成时蛋白浓度、加热温度、加热时间、蛋白溶液pH对凝胶质构均有一定的影响。采用正交实验设计,通过质构仪进行物性测定比较其凝胶性,大豆11S球蛋白浓度14%、加热温度90℃、溶液pH为7、加热时间60min条件下形成的凝胶强度最好,其凝胶面积410.30g·s,凝胶力117.9g。      

Effect of debranching on the rheological properties of Ca-pectin gels

Water-soluble pectin (WSP) extract was subjected to controlled carrot pectin methylesterase treatment, thereby producing de-esterified pectin (DEP). Both WSP and DEP were incubated with a mixture of endo-arabinanase and α-L-arabinofuranosidase to yield partially debranched pectins (WSP_DBr and DEP_DBr respectively). Pectin samples were characterised in terms of degree of methylesterification (DM), neutral sugar content, and degree of branching (DBr). The characterised pectins were used for the preparation of pectin gels with high and low calcium ion (Ca²⁺) concentrations. The rheological characteristics of the produced gels were evaluated by means of small-amplitude oscillatory tests. These characteristics include network development of the gel, gel strength (G′), gel elastic character and gel type. Partial debranching of pectin resulted in a reduction of the arabinose content (by approximately 50%) and caused a slight decrease in polymer DBr. Gels produced from semi-dilute solutions of partially debranched pectins showed network development profiles similar to those prepared from semi-dilute solutions of the corresponding non-debranched polymers. Yet, the former gels showed lower G′ values, poor gel elastic character and a “weaker” nature as compared to the latter. Next to Ca²⁺ cross-links, the presence of long arabinose-containing side chains is suggested to play an important role in the rheological characteristics of Ca²⁺-pectin gels.     

Analysis of fish and squid myofibrillar proteins by capillary sodium dodecyl sulfate gel electrophoresis: actin and myosin quantification

 A capillary electrophoretic method for the separation and quantification of fish and squid myofibrillar proteins was developed. The method uses sodium dodecyl sulfate and β-mercaptoethanol for solubilization and analysis of myofibrillar protein subunits. The separation of the different polypeptides is achieved by the sieving effect of the gel inside the capillary. A calibration curve for myosin heavy chain and actin UV absorbance quantification of these proteins was developed. Received: 2 November 1999 / Revised version: 16 February 2000     

Effect of pH on the gel properties and secondary structure of fish myosin

The relationships between gel properties and the secondary structures of silver carp myosin were investigated at pH 5.5–9.0 using dynamic rheological measurement, circular dichroism and scanning electron microscopy. The gel properties of fish myosin were strongly pH and temperature dependent. During heating at 1 °C/min, myosin formed gels in the pH range 5.5–7.5, but not at pH 8.0–9.0. α-Helix was the predominant structure at pH 7.0. The α-helix fraction declined with increasing temperature and the pH away from 7.0, whilst the other secondary structure fractions increased. The α-helix structure of myosin was more susceptive to acid-treatment than alkali-treatment. As pH increased, the gelation rate and gel strength decreased, and the water-holding capacity (WHC) showed an increasing trend followed by a plateau. High β-sheet and β-turn fractions prior to heating could improve G′ at 90 °C, but they depressed the WHC. A compact and uniform gel of fish myosin was obtained at pH 7.0.