Protein Denaturation,Rheology, and Gelation Characteristics of Radio-Frequency Heated Egg White Dispersions

Gel properties of radio frequency (RF) heated egg white dispersions at 27.12 MHz were studied as function of concentration (2.5–2.5 kg/100 kg sample), pH (3–11) and heating time (60–180 s). Egg white dispersions demonstrated a gradual liquid-solid transformation as they denatured and gelled during RF treatment. The critical concentration and heating period for egg white protein denaturation and gelation were found to be 7.5% (w/w) and 150 seconds. The elastic modulus (G′) of RF-heated samples increased with concentration and heating period (temperature), whereas complex viscosity (η*) increased exponentially with concentration. In an alkaline condition, the egg white dispersion did not produce a gel; however, in acidic condition it resulted in a strong gel with significantly (P?<?0.05) higher G′. This could be attributed to the high dielectric constant (ε′) and loss factor (ε″) values of acidified samples as compared to the alkaline and control egg white dispersion. Effect of heating rate (1, 5, 10, and 20°C/min) in situ on rheometer plate significantly affected gel rigidity; the RF treated sample rigidity was comparable to samples heated at the rate of 5 and 10°C/min. Differential scanning calorimetry, dielectric measurement, and sodium dodecyl sulfate (SDS) PAGE electrophoresis results were used to confirm gelation behavior during both conventional and RF heating conditions.     

Effects of inulin/oligofructose on the thermal stability and acid-induced gelation of soy proteins

ABSTRACT:  Differential scanning calorimetry (DSC) and dynamic oscillatory shear testing were performed to study the influence of inulin (Raftiline^® HP-gel and Raftiline^® ST-gel) and oligofructose (Raftilose^® P95) on the thermal stability and gelation (using glucono-δ-lactone [GDL] as a coagulant) of soy protein isolate (SPI) dispersions. Addition of 10% (w/v) inulin/oligofructose or sucrose increased ( P < 0.05) the peak denaturation temperatures ( T_m ) of 7S and 11S soy proteins in SPI dispersion (5%[w/v], pH 7.0) by an average of 1.9 and 2.3 °C, respectively. GDL induced SPI thermal gelation, and the gel rheology was affected by both the pH decline and the specific temperature of heating. Addition of inulin/oligofructose (8%, w/v) improved the gelling properties of preheated SPI dispersion (8%, w/v) coagulated with GDL, showing 14.4 to 45.6% increase ( P < 0.05) in gel rigidity ( G ' value) at the end of heating (81 °C). Microstructural examination revealed a denser protein cross-linking structure and reduced pore sizes in SPI gels containing inulin/oligofructose. In general, inulin was more capable of improving SPI gelation than oligofructose, suggesting that the degree of fructose polymerization in the fructans was of thermal and rheological importance.     

大豆分离蛋白对肌原纤维蛋白加热过程中结构及流变特性的影响

大豆分离蛋白（soybean protein isolate,SPI）作为优质的植物蛋白常被用于肉制品加工中,以提高产品产量和质地。研究添加SPI对肌原纤维蛋白（myofibrillar protein,MP）凝胶特性及MP加热过程中结构和流变特性的影响。结果表明：添加10%、20% SPI能提升混合凝胶的凝胶强度及保水性（P＜0.05）;加热过程中混合蛋白凝胶二级结构发生改变,但其变化规律尚不明确;添加SPI使混合凝胶储能模量及损耗模量下降;混合凝胶上清液十二烷基硫酸钠-聚丙烯酰胺凝胶电泳图谱显示,肌球蛋白重链、肌动蛋白、SPI部分亚基均是参与凝胶形成的蛋白质。     

Molecular interactions and functionality of a cold-gelling soy protein isolate

ABSTRACT:  A soy protein isolate (SPI) was thermally denatured at a critical concentration of 8% protein for 3 h at 95 °C, resulting in a powder that was readily reconstituted at ambient temperature and that demonstrated improved heat stability and cold-set gel functionality when compared to a control SPI. When SPI was heated at 3% protein equivalently, prior to reconstitution to 8% protein, the final viscosity was about 3 orders of magnitude less than the original sample. The viscosity of SPI heated at 3% protein was still nearly 2 orders of magnitude less than the original sample after both samples were reheated at 8% protein. These results suggested that heat denaturation at low protein concentrations limited network formation even after the protein concentration and interaction sites increased, impacting the isolate's cold gelling ability. Gelation was prevented upon treatment of SPI with iodoacetamide, which carbaminomethylated the cysteine residues, establishing the role of disulfide bonds in network formation. The viscosity of the 8% protein dispersion was also reduced by 2 orders of magnitude when treated with 8 M urea, and when combined with 10 mM DTT the gel viscosity was decreased by another order of magnitude. These results suggested that hydrophobic interactions played a primary role in gel strength after disulfide bonds form. The need for a higher concentration of protein during the heating step indicated that the critical disulfide bonds are intermolecular. Ultimately, the functionality produced by these protein–protein interactions produced a powdered soy protein isolate ingredient with consistent cold-set and thermal gelation properties.     

谷氨酰胺转氨酶预交联改善CaSO4诱导大豆分离蛋白凝胶性的研究

酶法改性能够有效提升大豆蛋白的凝胶性。为了探讨谷氨酰胺转氨酶(transglutaminase, TGase)预交联对盐诱导大豆分离蛋白凝胶性的影响,通过控制酶浓度、预交联时间制备不同预交联程度的大豆分离蛋白(soy protein isolate,SPI)溶液,并研究其在CaSO₄作用下的成胶性能。结果显示,与未经TGase处理的SPI相比,TGase适度预交联能够显著提升SPI的凝胶品质。经3～5 U/g TGase预交联20 min或3 U/g TGase预交联20～30 min后,SPI凝胶性得到了不同程度的提升,其中弹性模量、屈服应力、屈服应变、持水率最大分别提高了124.5%、269.0%、135.0%及53.0%。然而,过度预交联产生过大的蛋白聚集体,导致最终形成的凝胶结构粗糙、多孔,凝胶强度、持水力等均显著下降(P<0.05)。由此可见,合理利用TGase对蛋白进行预交联处理能够改善SPI凝胶制品品质,对于TGase在食品工业中的应用及传统豆制品质构改良具有重要的指导意义。     

Influences of Gums, Soy Protein Isolate, and Heating Temperatures on Reduced-Fat Meat Batters in a Model System

Effects of incorporation of iota-carrageenan, sodium alginate (AL), and soy protein isolate (SPI) in comparison with control (CONT) on the textural and physicochemical properties of reduced-fat (15%) meat emulsions heated to various internal temperatures were investigated. Lower percent fat and water losses for treatments containing gums or soy protein isolate than those of CONT indicated that addition of gums and soy protein isolate could improve emulsion stability. AL had the greatest water-holding capacity at the higher endpoint temperatures (76.7 °C and 82.2 °C), which was possibly due to the formation of a heat-stable alginate gel. The SPI had the highest concentration of salt-soluble protein in either raw or cooked meat batter.     

pH Induced Aggregation and Weak Gel Formation of Whey Protein Polymers

Whey protein polymers were formed by heating (80 °C) a 4% (w/v) whey protein (WP) isolate dispersion at pH 8.0 for 15, 25, 35, 45, or 53 min. Dispersions were adjusted to pH 6.0, 6.5, 7.0, 7.5, or 8.0 after heating and the rheological properties were determined. Viscosity increased with increased heating time and decreased pH. At pH 7.0 and 7.5, high-viscosity dispersions with pseudoplastic and thixotropic flow behavior were formed, while weak gels were formed at pH 6.0 and 6.5. The storage (elastic) and loss (viscous) moduli of pH-induced gels increased when temperature was increased from 7 °C to 25 °C, suggesting that hydrophobic forces are responsible for gelation. Key Words: weak-gels, whey proteins, polymers, gelation, functionality     

Gelation kinetics of an imitation-mortadella emulsion during heat treatment determined by oscillatory rheometry

The gelation kinetics of an imitation-mortadella emulsion during thermal treatment under both, isothermal and non-isothermal conditions was followed by means of oscillatory rheometry. A first order-kinetic process was found to better describe the variation of the storage modulus G′ (at 1 Hz and strain amplitude of 0.8%) during the two types of thermal treatments. An Arrhenius-type model was able to describe the effect of temperature during gelation of the emulsion under non-isothermal conditions.     

Thermal syneresis affected by heating schedule and moisture level in surimi gels

ABSTRACT:  The extent of thermal syneresis in protein gelation is indicative of thermal and freeze-thaw stability as well as the network integrity of a protein gel. Thermal syneresis in Alaska pollock surimi gels was examined under different heating schedules (40 °C/20 min to 90  °C/30 min, 60 °C/20 min to 90 °C/30 min, and 90 °C/20 min to 90 °C/20 min) at varying moisture levels (80%, 82%, and 84%). The extent of syneresis and gel firming was monitored by centrifugation expressible moisture and penetration force, respectively. The occurrence of 2 distinct peaks as a function of time for both thermal syneresis and gel firming suggests that a multistage aggregation is involved in the formation of gel network. All syneresis preceded gel firming upon protein aggregation. Increasing the moisture content in the gel delayed the 2nd stage of protein aggregation. The 60 °C/20 min preheating followed by 90 °C/30 min postheating resulted in significantly greater thermal syneresis and gel weakening compared to 40 and 90 °C preheating. Changes of gel structure clearly reflected thermal syneresis when the size of water pores became smaller with initiation of network formation and progressively larger upon further heating. Thermal syneresis history during protein gelation can be used to predict thermal and freeze-thaw stability.     

Rheological Studies on Gelling Behavior of Soy Protein Isolates

The gelling behavior of soy protein isolate (SPI) dispersions with solid contents 0.8-8.8% was studied by a dynamic fluid rheometer. Strain sweep and frequency sweep measurements showed the dispersions were stable and predominantly elastic at room temperature. Elasticity increased sharply when dispersions were heated. Cooling set was found for dispersions of higher solid contents. For solids contents < 3%, cooling set was not as pronounced; and cooling could result in reduced elasticity when the prior heating was milder. Results suggest SPI changed from an aqueous dispersion to a dispersion consisting of particles embedded in heat-soluble protein sol matrix upon heat treatment, and further underwent a transition into a gel consisting of particles embedded in heat-formed protein gel matrix. Thermoreversibility of elasticity was determined by solids content.     

Effect of acid hydrolysis on rheological and thermal characteristics of lentil starch slurry

A comparative rheological and thermal study was carried out between acid hydrolyzed and unhydrolyzed (control) lentil starch dispersions (25-33.3 g starch per 100 g water) as function of temperature. After acid hydrolysis, the peak gelatinization temperature (T_p) shifted to higher temperature than the corresponding starch without hydrolysis whereas the gelatinization enthalpy remained unaffected by hydrolysis. The starch gelatinization kinetics was evaluated by a non-isothermal technique as function of elastic modulus (G′) and G′ vs. time (t) data up to the gelatinization peak value was considered for rate estimation. A 2nd-order reaction kinetics described well the starch gelatinization process and the process activation energy was ranged between 241 and 434 kJ/mol. Acid hydrolysis strongly affected the rheological properties by lowering gel strength compared to unhydrolyzed starch. The creep analysis further revealed that starch gel was significantly affected by hydrolysis and exhibited less resistant to the stress. A 4-parameters Burgers model well-described creep curves and supported oscillatory rheological data.     

β-Lactoglobulin Gelation and Modification: Effect of Selected Acidulants and Heating Conditions

ABSTRACT: The effect of acidulant selection, heating temperature, and heating rate on the properties of low-pH β-lactoglobulin (β-Lg) gels and powders derived from these gels was investigated by rheological and microscopic techniques. As isothermal gelation temperature was increased from 75 to 85 °C, gels made with hydrochloric and lactic acid showed more rapid gel formation and increased stress at gel fracture. Thickening and water-holding properties of powders derived from these gels also increased with temperature. Increases in gel strength and derivatized powder functionality appeared to plateau above 85 °C. Gels and derivatized powders prepared with phosphoric acid exhibited attributes similar to samples prepared with HCl and lactic acid at lower temperatures. The ion-specific ability of phosphate to increase denaturation temperature was responsible for the shift in properties of gels made with phosphoric acid. Microscopy revealed temperature effects on network building block size, but variations in rheological properties could not be linked to changes in gel micrographs. Alteration of heating rates from 2.0 to 0.2 °C/min during gelation affected the observed gelation temperature, but had little effect on final gel mechanical properties. Acid selection and gelation temperature offer alternatives to control β-Lg gel strength and the functional properties of instant thickening protein ingredients.     

Physicochemical properties of soy protein isolate gels emulsified with various oils using a microbial transglutaminase

Soy protein isolate (SPI) gels emulsified with oils including soybean, olive, palm, and eicosapentaenoic acid (EPA) were prepared by a microbial transglutaminase (MTGase). The hardness of 10% SPI gel was greatly increased by adding higher amount of oils. The emulsion gels prepared with 10% SPI and 30% olive oil showed the highest hardness of 1,711 g. In the gelation with various oil content (5–30%), the higher concentration of oils indicated the drastic increase of elastic modulus G′ and viscous modulus G″ during initial gelation for 7 min. The G′ value of SPI emulsion gel showed the 150 (soybean oil 30%), 147 (olive oil 30%), 121 (palm oil 30%), and 61 Pa (EPA 25%), respectively. In the color value of SPI emulsion gel, addition of higher concentration of oils resulted in the increase of L value (brightness), indicating 99.14 (L value) at 30% palm oil. The micro-structure of SPI emulsion gel entrapped with various oils showed the homogeneous network with small porosity compared with that of SPI gel without oil. In particular, SPI emulsion gel with 10% palm oil showed the compact structure distributed evenly with small porosity. Conclusively, the functional and rheological properties of SPI emulsion gel produced by catalytic action of MTGase could be modulated by the type and content of oils fortified.     

Gel Properties of Surimi from Bighead Carp (Aristichthys nobilis): Influence of Setting and Soy Protein Isolate

ABSTRACT: The effects of setting conditions and soy protein isolate (SPI) on textural properties and microstructures of surimi produced from bighead carp were investigated. The incubation conditions of bighead carp surimi affected the breaking force and distance. The optimum setting conditions were 35 °C to 40 °C for 60 min. When the surimi was cooked after 50 °C incubation for 30 to 120 min, the breaking force and distance were inferior to that of no incubation. The gel structure showed that the incubation conditions affected the bighead carp surimi gel microstructures, thus producing surimi with different gelling properties. Breaking force and distance of surimi gels decreased when the protein ratio of SPI was increased in the total protein at 30 °C and 40 °C for 60 min setting and heating at 85 °C for 30 min, but the breaking force obtained for 90% surimi protein plus 10% SPI protein was higher than surimi alone at 50 °C for 60 min incubation and heating at 85 °C for 30 min.     

THE FORMATION OF HEAT AND ENZYME INDUCED (PLASTEIN) GELS FROM PEPSIN-HYDROLYZED SOY PROTEIN ISOLATE1

A gel prepared by heating pepsin-hydrolyzed soy protein isolate (HSPI) had a higher turbidity and viscosity and less solubility than did its starting hydrolyzate. The gelling characteristics of the heated hydrolyzate were possibly due to hydrophobic interactions, as opposed to peptide bond reformation. The presence of pepsin was found to be necessary for gel formation at 37°C but could be successfully replaced by heat. Incubation of the concentrated HSPI at 95°C for 30 min was optimal for heat-induced gel formation. When soy protein isolate (SPI) was hydrolyzed with papain or bacterial protease, little gelation occurred (either by heat or enzyme induced gelation treatments).     

Acid-induced gelation of whey protein polymers: effects of pH and calcium concentration during polymerization

Heating whey protein dispersions (90°C for 15 min) at low ionic strength and pH values far from isoelectric point (pH>6.5) induced the formation of soluble polymers. The effect of mineral environment during heating on the hydrodynamic characteristics and acid-induced gelation properties of polymers was studied. Whey protein dispersions (80 g/l) were denatured at different pH (6.5–8.5) and calcium concentrations (0–4 mm) according to a factorial design. At pH 6.5, the hydrodynamic radius of protein polymers increased with increasing calcium concentration, while the opposite trend was observed at pH 8.5. Intrinsic viscosity results suggested that heating conditions altered the shape of protein polymers. Whey protein polymers were acidified to pH 4.6 with glucono-δ-lactone and formed opaque particulate gels. The storage modulus and firmness of gels were both affected by conditions used to prepare protein polymers. As a general trend, polymers with high intrinsic viscosity produced stronger gels, suggesting a relationship between polymer shape and gel strength.Acid gelation properties of whey protein polymers makes them suitable ingredients for yoghurt applications. Using whey protein polymers to standardize protein content increased yoghurt viscosity to 813 Pa.s while using skim milk powder at same protein concentration increased yoghurt viscosity to 393 Pa.s. Water holding capacity of protein polymers in yoghurt was 19.8 ml/g compared to 7.2 ml/g for skim milk powder protein. Acid gelation properties of whey protein polymers are modulated by calcium concentration and heating pH and offers new alternatives to control the texture of fermented dairy products.     

Gelation Kinetics of Agars from Pterocladia capillacea Examined by Dynamic Rheometry

ABSTRACT: The changes in storage modulus (G') during isothermal gelation of agar fractions from Pterocladia capillacea were kinetically investigated with respect to the curing condition, agar concentration, and the molecular property of agar. Generally, the reciprocal of gelling time, the maximal increasing rate of G', and the rate constants of Avramipseudo-1st-order kinetics correlated with the curing conditions more significantly than with the molecular properties examined. This tendency was opposite to the result based on simple 2nd-order rate constant. And, the effects of molecular size on rheological and kinetic parameters were greater than those of the 3,6-anhydro-galac-tose content examined. After verification of the kinetic data obtained, potentially molecular changes during gelation process are illustrated.     

The effect of transglutaminase crosslinking on the rheological characteristics of heated peanut flour dispersions

ABSTRACT:  Peanut flour (PF) is a high-protein ingredient prepared after the partial extraction of oil from roasted peanut seed. Microbial transglutaminase (TGase) catalyzes protein crosslinking via acyl-transfer reactions, resulting in the modification of functional properties such as viscosity, gelation, solubility, and water holding capacity. This work was conducted to observe changes in rheological properties of PF dispersions in the presence and the absence of TGase and amidated pectin (AP). Dispersions were characterized across a range of conditions, including controlled heating and cooling rates under both large- and small-strain deformations. Gelation occurred at temperatures above 78 °C using PF dispersions treated with TGase compared to untreated dispersions devoid of the enzyme (about 68 °C). The addition of AP (0.5%) resulted in a general increase in viscoelasticity for all dispersions. AP addition also minimized the shift in gel point temperature caused by TGase polymerization reactions. High-molecular-weight polymers were formed in TGase-treated PF dispersions in both the presence and the absence of AP; however, polymer formation was more rapid in PF dispersions without AP. Ortho-phthaldialdehyde assays indicated about 40% protein coupling in PF dispersions treated with TGase compared to about 20% in those containing both AP and TGase. Collectively, these data suggest potential applications of TGase-treated PF dispersions, both in the presence and the absence of AP, for use in peanut-base food products, including protein bars, shakes, and value-added baked goods.     

Gelation Characteristics of Paddlefish (Polyodon spathula) Surimi Under Different Heating Conditions

Gelation properties of paddlefish surimi were investigated with different heating procedures. Without pre-incubation, gel strength of paddlefish surimi increased as temperature increased from 40 to 60 °C. Pre-incubation at 40 °C caused myosin degradation and reduced gel strength by 55% compared to the control. Pre-incubation at 70 °C followed by cooking at 90 °C produced gels with maximum strength. Isothermal heating between 40 and 50 °C produced rheological transitions between 0 and 15 min. Beef plasma powder reduced myosin degradation and enhanced gelation of surimi incubated around 40 °C. These results indicated that the gel-weakening phenomenon in paddlefish surimi was due to the degradation of myosin by some endogenous protease(s).     

Elucidation of structural changes in soy protein isolate upon heating by Raman spectroscopy

Changes in protein secondary and tertiary structure of soy protein isolate (SPI) upon heating process were investigated by Raman spectroscopy. SPI refrigerated and heated at 70, 100 and 200 °C was analysed. Non-significant differences in secondary structure (α-helix, β-sheet, unordered and turn) were observed in SPI upon heating at these temperatures. However, comparison of the Raman spectra of SPI refrigerated and heated at 70, 100 and 200 °C reveals changes in hydrophobic group environments. An intensity decreasing trend of the 1340 cm⁻¹ band attributable to tryptophan vibrations was observed, which involves an increase in solvent exposure of tryptophan residues. The spectral results also showed intensity increasing trends for the 1450 and 2935 cm⁻¹ bands of SPI upon heating, which can be ascribed to breaking of hydrophobic contacts and subsequent solvent exposure of the corresponding hydrophobic aliphatic groups. Finally, it was found tertiary structural transitions towards greater proportions of buried tyrosine residues upon heating of SPI.