Pilot-scale formation of whey protein aggregates determine the stability of heat-treated whey protein solutions—Effect of pH and protein concentration

Denaturation and consequent aggregation in whey protein solutions is critical to product functionality during processing. Solutions of whey protein isolate (WPI) prepared at 1, 4, 8, and 12% (wt/wt) and pH 6.2, 6.7, or 7.2 were subjected to heat treatment (85°C × 30 s) using a pilot-scale heat exchanger. The effects of heat treatment on whey protein denaturation and aggregation were determined by chromatography, particle size, turbidity, and rheological analyses. The influence of pH and WPI concentration during heat treatment on the thermal stability of the resulting dispersions was also investigated. Whey protein isolate solutions heated at pH 6.2 were more extensively denatured, had a greater proportion of insoluble aggregates, higher particle size and turbidity, and were significantly less heat-stable than equivalent samples prepared at pH 6.7 and 7.2. The effects of WPI concentration on denaturation/aggregation behavior were more apparent at higher pH where the stabilizing effects of charge repulsion became increasingly influential. Solutions containing 12% (wt/wt) WPI had significantly higher apparent viscosities, at each pH, compared with lower protein concentrations, with solutions prepared at pH 6.2 forming a gel. Smaller average particle size and a higher proportion of soluble aggregates in WPI solutions, pre-heated at pH 6.7 and 7.2, resulted in improved thermal stability on subsequent heating. Higher pH during secondary heating also increased thermal stability. This study offers insight into the interactive effects of pH and whey protein concentration during pilot-scale processing and demonstrates how protein functionality can be controlled through manipulation of these factors.     

Effect of Dehydration Methods on the Functionality of Plant Protein Concentrates

The significance of different dehydration methods for rice bran protein and soy protein coagulates on composition and functionality of the resulting protein concentrates was investigated. Minor compositional differences were found between the freeze-, spray- or drum-dried samples. Protein solubility data indicated limited effects of dehydration methods on heat coagulated samples but significant reduction of protein solubility by drum drying was found in the case of soy protein coagulated at room temperature. Water absorption capacity and emulsion properties were significant affected by dehydration methods but no consistent trends could be observed for fat absorption capacity. A significant increase of specific bread loaf volume, relaxation time of doughs and in some cases deformation of protein fortified samples was detected from freeze-dried to drum-dried samples. In general the effects of dehydration methods on protein functionality were more consistent for heat coagulated rice bran protein than for soy protein concentrates processed at ambient temperatures.     

Differential Scanning Calorimetry Analysis of the Effects of Heat and Pressure on Protein Denaturation in Soy Flour Mixed with Various Types of Plasticizers

The effects of heat and pressure on protein denaturation in soy flour were explored by an experimental design that used pressure (atmospheric to 600 MPa), temperature (room to 90 °C), time (1 to 60 min), and type of aqueous plasticizer (NaCl, sucrose, betaine, and lactobionic acid (LBA)) as factors. When 50% (w/w) soy flour‐water paste was high hydrostatic pressure (HHP)‐treated for 20 min at 25 °C, the treatment at 200 MPa showed a small effect on denaturation of only the 7S soy globulin, but the treatment at 600 MPa showed a significant effect on denaturation of both the 7S and 11S soy globulins. The treatment at 60 °C showed a less‐pronounced effect on denaturation of the 11S globulin, even at 600 MPa, but that at 90 °C showed a similar extent of denaturation of the 11S globulin at 600 MPa to that at 25 °C. Chaotropic 2N NaCl, 50% sucrose‐, 50% betaine‐, or 50% LBA‐water solutions showed protective effects on protein denaturation during HHP treatment at 25 °C. Although LBA enhanced the extent of thermostability of soy protein less than did 2N NaCl, LBA exhibited better stabilization against pressure. The results from DSC analysis demonstrated that thermostable soy proteins were not always barostable.     

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.     

Comparing the heat stability of soya protein and milk whey protein emulsions

The heat stability of emulsions stabilized by WPC or SPI or mixtures of the two are compared by following the change in oil droplet number during heating, and applying kinetic rate equations to calculate the rate constant (k) for destabilization. SPI emulsions were found to be unstable to heat at pH around the pI, whilst being stable at pH further from the pI. This is related to the pH dependent solubility of soy proteins. This determined that a pH close to the pI (pH 4.5) be used for further studies so as to give a heat labile emulsion. Both WPC and SPI emulsions showed a weak dependence of k on protein concentration at pH 4.5, and an increasing k as the temperature increased. Arrhenius plots for emulsions made with WPC were bilinear, whilst those for SPI followed a single straight line. The change in slope of the Arrhenius plots for the WPC emulsions occurred around 70 °C, lower than would be expected from the denaturation temperature of β-lactoglobulin, the protein that dominates the thermal behaviour of WPC. The activation energies for WPC and SPI emulsions calculated from the slopes of the Arrhenius plots are slightly lower for WPC and considerably lower for SPI than the equivalent values in the literature for these proteins in solution. This, and the apparent lower denaturation temperature of β-lactoglobulin in emulsions, we explain by hypothesizing that the WPC and SPI proteins are already partially denatured by surface adsorption when they are heated, and thus require less energy to denature, and unfold at lower temperatures than native non-adsorbed proteins.     

预热变性程度对大豆蛋白凝胶性质的影响

以非还原SDS-PAGE表征大豆蛋白的预热变性程度,并采用质构分析、流变分析研究预热变性程度对大豆蛋白凝胶性质的影响。SDS-PAGE结果表明,随着加热温度的升高蛋白质变性速率逐渐增大,相同加热时间条件下,蛋白质变性程度随加热温度升高呈S型曲线上升。质构、流变分析结果表明:随着预热变性程度的增大,大豆蛋白凝胶硬度先增大后减小,变性程度为86.11%时凝胶硬度最大,是未经预热变性的大豆蛋白凝胶硬度的2.15倍;凝胶弹性则随预热变性程度的增大持续增大;变性程度在22.28%以上时大豆蛋白形成凝胶更快,但完全变性大豆蛋白在形成凝胶时的降温阶段不能形成很好的凝胶结构。粒径分析结果表明,预热变性程度对大豆蛋白凝胶性质的影响与蛋白质预热变性时形成的聚集体尺寸及形态有关。     

FUNCTIONALITY COMPARISON BETWEEN DERIVATIZED WHEY PROTEINS AND A PREGELATINIZED STARCH

A process has been patented to produce stabilizing ingredients from whey proteins which are applicable over a wide range of typical food conditions and do not require heat or the addition of salts to induce thickening functionality. Once reconstituted in deionized water, solutions were evaluated and compared with water holding performance and rheological attributes of a pregelatinized cornstarch. Rotational viscometry was performed at pH values between 3 and 8, temperatures between 5 and 90C, and shear rates between 1 and 100 s⁻¹. Derivatized whey protein powders and pregelatinized starch displayed pseudo-plastic behavior under shear at all temperatures tested. During temperature ramps from 5 to 90C, derivatized whey protein flow properties were essentially unchanged by varying pH. However, viscosity after the temperature increase was higher than initial values, possibly due to additional protein denaturation and hydrophobic interactions. Derivatized powders were stable and retained desired functionality over a wide range of food processing and preparation conditions and may therefore possess applicability to many products currently utilizing modified starches or hydrocolloids to texturize.     

不同热处理条件下大豆分离蛋白的红外光谱分析

本文研究不同温度、时间热处理下不同浓度大豆分离蛋白的热稳定性及红外光谱,探讨蛋白质二级结构的变化规律,结果表明:样品浓度的差异对变性温度（T_D）和变性焓变（ΔH）的影响不明显,未经加热处理的大豆分离蛋白中7S和11S球蛋白变性温度分别为74.2℃和93.7℃。相比于未处理的蛋白质样品,热处理使α-螺旋结构增多,β-折叠结构减少。80℃热处理下,α-螺旋结构及β-转角结构均呈现先增加后减少的变化趋势,而β-折叠结构含量则先减少后增加。在2%蛋白浓度、90℃热处理条件下,随着处理时间的增长,α-螺旋结构及β-折叠结构呈现出先增加后降低的变化趋势,而无规卷曲结构则呈现相反的变化趋势。无论何种蛋白浓度下,11S球蛋白在90℃热处理45 min时的变性增大了无规卷曲结构含量,同时降低了β-转角结构及β-折叠结构组分含量。      

Thermal behavior of emulsions manufactured with soy protein ingredients

The conformation, denaturation and aggregation behavior of proteins are important factors which dictate their ingredient functions and applications in formulated food products. The effect of variation in pre-treatment temperature (70–90 °C × 30 s), pH (6.4–7.5) and calcium supplementation (450 and 850 mg/L) on heat coagulation time (HCT at 140 °C) of model emulsions (3.6% (w/v) protein) stabilized with soy protein isolate (SPI) and soy protein hydrolysate (SPH) ingredients was determined. Generally, HCT of emulsions was not significantly affected by alteration of constituent pre-heating temperatures. Model emulsions displayed higher HCTs with increasing pH and lower levels of intrinsic ash content. At both supplementation levels, calcium addition led to decreased HCTs. Supplementation with chloride salts caused a greater decrease in HCT compared to supplementation with citrate salts. Furthermore, soy protein hydrolysis was associated with lower emulsion thermal stability. Results demonstrate that modification of ingredient and manufacturing parameters may be a useful approach for enhancing thermal stability properties of soy protein stabilized emulsions.     

Gelation Property of Alcohol-Extracted Soy Protein Isolate and Effects of Various Reagents on the Firmness of Heat-Induced Gels

Gelation property, thermal property, protein subunits distribution, and molecular forces involved in gelation of alcohol-extracted soy protein isolate were investigated using texture analyzer, differential scanning calorimeter, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and various reagents. Results showed that salts and pH played important roles in gel firmness, and a power law relationship between gel firmness and protein concentration was observed. The effects of various reagents revealed that disulfide bonds play a major role in soy protein gel formation, while the involvement of electrostatic interactions, hydrogen bonds, and/or hydrophobic interactions also occurred in gel networks. Thermal analysis indicated that both alcohol-extracted soy protein isolate and commercial soy protein isolate (isoelectric precipitation) have undergone serious denaturation, while the gel firmness of alcohol-extracted soy protein isolate was significantly greater than that of commercial soy protein isolate. Sodium dodecyl sulfate polyacrylamide gel electrophoresis image showed that there was almost no difference for protein subunits among alcohol-extracted soy protein isolate, commercial soy protein isolate, and soy powder. Hence, as an alternative environmental friendly extraction method, alcohol-extraction of soy protein isolate has a great prospect to replace presently applied isoelectric precipitation method.     

红外加热对大豆蛋白质湿热变性的影响

实验以大豆粉为原料,利用红外加热方法,研究其对大豆蛋白质变性程度的影响,通过单因素及正交实验研究了热处理时间、热处理温度和固液比对大豆蛋白质溶解度有较大的影响。分析得出大豆蛋白质变性最佳条件,即加热温度为110℃,加热时间为35 min,固液比(w/w)为1∶2;在此条件下,大豆粉蛋白质的氮溶解指数为22.36%,即大豆蛋白质的变性最高。      

Effect of soybean addition on the rheological properties and breadmaking quality of wheat flour

The effects of enzyme‐active full‐fat (EAFFSF), heat‐treated full‐fat (HTFFSF) and enzyme‐active defatted (EADSF) soy flours and commercial soy protein isolates (SPIs) on mixing properties and extensibility of dough, gluten formation, gas production and retention properties of dough and bread quality were studied. The soy products utilised in this study presented values of urease activity, nitrogen solubility index (NSI) and enthalpies of protein denaturation according to their previous heat processing. Soy products interfered in gluten formation, weakened dough strength and decreased dough gas retention capacity. Bread quality was negatively affected by soy product addition. The negative effects were exacerbated by an increase in soy/wheat ratio. Soybean protein state was identified as an important factor in determining dough and bread properties. Copyright © 2005 Society of Chemical Industry     

Effect of pH and heat treatment conditions on physicochemical and acid gelation properties of liquid milk protein concentrate

Milk protein concentrates (MPC) are typically dried high-protein powders with functional and nutritional properties that can be tailored through modification of processing conditions, including temperature, pH, filtration, and drying. However, the effects of processing conditions on the structure-function properties of liquid MPC (fluid ultrafiltered milk), specifically, are understudied. In this report, the pH of liquid MPC [13% protein (70% protein DM basis), pH 6.7] was adjusted to 6.5 or 6.9, and samples at pH 6.5, 6.7, and 6.9 were subjected to heat treatment at either 85°C for 5 min or 125°C for 15 s. Sodium dodecyl sulfate PAGE was used to determine the distribution of caseins and denatured whey proteins in the soluble and micellar phases, and HPLC was used to quantify native whey proteins as a measure of denaturation, based on the processing conditions. Both heat treatments resulted in substantial whey protein denaturation at each pH, with β-lactoglobulin denatured more extensively than α-lactalbumin. Changes in liquid MPC physicochemical properties were monitored at d 1, 5, and 8 during storage at 4°C. Viscosity increased after heat treatment and also over time, regardless of pH and heating conditions, suggesting the role of whey protein denaturation and aggregation, and their interactions with casein micelles. The MPC samples processed at pH 6.9 had a significantly higher viscosity than those heated at pH 6.5 or 6.7, for both temperature and time conditions; and samples processed at 85°C for 5 min had higher viscosity than those heated at 125°C for 15 s. Particle size analysis indicated the presence of larger particles after 5 and 8 d of MPC storage after heating at pH 6.9. Acid-induced gelation of the liquid MPC led to significantly higher gel firmness after processing at 85°C for 5 min, compared with 125°C for 15 s. Also, gels made from MPC adjusted to pH 6.5 had higher storage moduli, with both time and temperature combinations, demonstrating the role of pH-dependent association of denatured whey proteins with casein micelles in gel network formation. These findings enable a better understanding of the processing factors contributing to structural and functional properties of liquid MPC and can be helpful in tailoring milk protein ingredient functionality for a variety of food products.     

Physicochemical and Functional Properties of Winged Bean Flour and Isolate Compared with Soy Isolate

The proximate composition, amino acid profile and functional properties of isolated winged bean proteins were determined and compared with soy protein isolate. Winged bean protein extracted at pH 10 and pH 12 had protein contents of about 90% and 80%, respectively. Alkali extraction of winged bean proteins at pH 10 and pH 12 did not affect the amino acid distribution of the isolated proteins. Oil and water absorption, emulsion, and foaming properties of winged bean isolated compared favorably with soy isolate. Least gelation concentration for winged bean isolate was 18% compared to 14% for soy isolate. Thus, winged bean protein isolate with its high protein content, high lysine and other essential amino acid content and good functionality has a good potential as an ingredient in food products.     

Effects of different oils on the properties of soy protein isolate emulsions and gels

The emulsion properties, protein adsorption and visco-elastic properties of heat-treated soy protein isolate (SPI) emulsions containing sunflower oil, soy oil and palm stearin were studied at neutral pH. The rheological properties, textural profile and fat adsorption capacity of subsequently induced glucono-δ-lactone (GDL) gels were investigated. At neutral pH the palm stearin emulsions showed higher stability than the sunflower and soy oil emulsions. The former also showed higher protein adsorption, emulsifying activity index and visco-elastic properties than the latter, indicating a higher denaturation degree for SPI at the oil/water interface of palm stearin emulsions at neutral pH. Acidified SPI-palm stearin emulsion gels were significantly harder than the soy and sunflower oil versions as well as the control (SPI without oil), whereas gels containing the two liquid oils were softer than the control. SPI-palm stearin gels also had higher fat adsorption capacity than SPI-sunflower oil/soy oil emulsion gels.     

Modifying the Functional Properties of Egg Proteins Using Novel Processing Techniques: A Review

Egg proteins can be used in a wide range of food products, owing to their excellent foaming, emulsifying, and gelling properties. Another important functional property is the susceptibility of egg proteins to enzymatic hydrolysis, as protein digestion is closely related to its nutritional value. These functional properties of egg proteins are likely to be changed during food processing. Conventional thermal processing can easily induce protein denaturation and aggregation and consequently reduce the functionality of egg proteins due to the presence of heat‐labile proteins. Accordingly, there is interest from the food industry in seeking novel nonthermal or low‐thermal techniques that sustain protein functionality. To understand how novel processing techniques, including high hydrostatic pressure, pulsed electric fields, ionizing radiation, ultraviolet light, pulsed light, ultrasound, ozone, and high pressure homogenization, affect protein functionality, this review introduces the mechanisms involved in protein structure modification and describes the structure–functionality relationships. Novel techniques differ in their mechanisms of protein structure modification and some have been shown to improve protein functionality for particular treatment conditions and product forms. Although there is considerable industrial potential for the use of novel techniques, further studies are required to make them a practical reality, as the processing of egg proteins often involves other influencing factors, such as different pH and the presence of other food additives (for example, salts, sugar, and polysaccharides).     

Sensory and functional properties of cheese: incorporation of whey proteins by pH manipulation and heat treatment

The effect of pH manipulation prior to high heat treatment on the effectiveness of whey protein denaturation in milk for cheesemaking has been studied. It has been found that the severity of heat treatment of milk required for denaturation can be reduced if the pH is altered during heating. The sensory and functional properties of cheddar-type cheese produced for high heat treated milk can be improved by manipulation of pH on heat treatment of milk.     

大豆分离蛋白性能优化关键技术

大豆分离蛋白的性能包括相容性和功能性,相容性决定了大豆蛋白的应用范围,功能性则最终反映大豆蛋白用户的收益大小,通过醇洗法脱除异味,选择性分离分级,控制蛋白质变性,蛋白质改性,以及干燥后处理可以改善大豆分离蛋白的相容性和功能性,并对它们的原理和方法进行了探讨。     

Effects of temperature on food proteins and its implications on functional properties

This article surveys the knowledge in the area of protein structure and chemistry of denaturation prior to an indepth review of the effects of heat on soy, milk, and egg proteins. It also reviews the methods available to assess denaturation of proteins. Protein denaturation is an ambiguous phenomenon and the consequences of denaturation on the functional properties of proteins is further confounded by this ambiguity. For each of the three food proteins, the known chemistry of individual proteins is reviewed followed by observations made on changes induced by heat in each protein group. Food proteins are not pure entities and purification and physicochemical characterization of various components of the food proteins have not been thoroughly investigated. Further, food is a complex milieu of water, fat, carbohydrate, vitamin, minerals, etc. along with proteins, and processing affects not only each individual component in the food but also the nature and intensity of intercomponent interactions in a food.     

Effects of milk heat treatment and solvent composition on physicochemical and selected functional characteristics of milk protein concentrate

Milk protein concentrate (MPC) powders (～81% protein) were made from skim milk that was heat treated at 72°C for 15 s (LHMPC) or 85°C for 30 s (MHMPC). The MPC powder was manufactured by ultrafiltration and diafiltration of skim milk at 50°C followed by spray drying. The MPC dispersions (4.02% true protein) were prepared by reconstituting the LHMPC and MHMPC powders in distilled water (LHMPC_w and MHMPC_w, respectively) or milk permeate (LHMPC_p and MHMPC_p, respectively). Increasing milk heat treatment increased the level of whey protein denaturation (from ～5 to 47% of total whey protein) and reduced the concentrations of serum protein, serum calcium, and ionic calcium. These changes were paralleled by impaired rennet-induced coagulability of the MHMPC_w and MHMPC_p dispersions and a reduction in the pH of maximum heat stability of MHMPC_p from pH 6.9 to 6.8. For both the LHMPC and MHMPC dispersions, the use of permeate instead of water enhanced ethanol stability at pH 6.6 to 7.0, impaired rennet gelation, and changed the heat coagulation time and pH profile from type A to type B. Increasing the severity of milk heat treatment during MPC manufacture and the use of permeate instead of water led to significant reductions in the viscosity of stirred yogurt prepared by starter-induced acidification of the MPC dispersions. The current study clearly highlights how the functionality of protein dispersions prepared by reconstitution of high-protein MPC powders may be modulated by the heat treatment of the skim milk during manufacture of the MPC and the composition of the solvent used for reconstitution.