Interactions of whey proteins during heat treatment of oil-in-water emulsions formed with whey protein isolate and hydroxylated lecithin

The interactions of proteins during the heat treatment of whey-protein-isolate (WPI)-based oil-in-water emulsions with and without added hydroxylated lecithin were studied by examining the changes in droplet size distribution and the quantity and type of adsorbed and unadsorbed proteins. Heat treatment at 90 degrees C of WPI emulsions resulted in an increase in total adsorbed protein; unadsorbed beta-lactoglobulin (beta-lg) was the main protein interacting with the adsorbed proteins during the first 10 min of heating, but after this time, unadsorbed alpha-lactalbumin (alpha-la) also associated with the adsorbed protein. In emulsions containing hydroxylated lecithin, the increase in total adsorbed protein during heat treatment was much lower and the unadsorbed beta-lg did not appear to interact with the adsorbed proteins during heating. However, the behavior of alpha-la during heat treatment of these emulsions was similar to that observed in the emulsions containing no hydroxylated lecithin. In the presence of NaCl, the particle size of the emulsion droplets and the quantities of adsorbed protein increased markedly during heating. Emulsions containing hydroxylated lecithin were less sensitive to the addition of NaCl. These results suggest that the binding of hydroxylated lecithin to unfolded monomers or intermediate products of beta-lg reduces the extent of heat-induced aggregation of beta-lg and consequently decreases the interactions between unadsorbed beta-lg and adsorbed protein. This was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of heated whey protein and hydroxylated lecithin solutions.     

Improving laccase catalyzed cross-linking of whey protein isolate and their application as emulsifiers

Whey protein isolate (WPI) was chemically modified by vanillic acid in order to enhance its cross-linkability by laccase enzyme. Incorporation of methoxyphenol groups created reactive sites for laccase on the surface of the protein and improved the efficiency of cross-linking. The vanillic acid modified WPI (Van-WPI) was characterized using MALDI-TOF mass spectrometry, and the laccase-catalyzed cross-linking of Van-WPI was studied. Furthermore, the vanillic acid modification was compared with the conventional approach to improve laccase-catalyzed cross-linking by adding free phenolic compounds. A small extent of the vanillic acid modification significantly improved the cross-linkability of the protein and made it possible to avoid color formation in a system that is free of small phenolic compounds. Moreover, the potential application of Van-WPI as emulsifier and the effect of cross-linking on the stability of Van-WPI emulsion were investigated. The post-emulsification cross-linking by laccase was proven to enhance the storage stability of Van-WPI emulsion.     

Lipid oxidation in corn oil-in-water emulsions stabilized by casein,whey protein isolate,and soy protein isolate

Proteins can be used to produce cationic oil-in-water emulsion droplets at pH 3.0 that have high oxidative stability. This research investigated differences in the physical properties and oxidative stability of corn oil-in-water emulsions stabilized by casein, whey protein isolate (WPI), or soy protein isolate (SPI) at pH 3.0. Emulsions were prepared with 5% corn oil and 0.2-1.5% protein. Physically stable, monomodal emulsions were prepared with 1.5% casein, 1.0 or 1.5% SPI, and > or =0.5% WPI. The oxidative stability of the different protein-stabilized emulsions was in the order of casein > WPI > SPI as determined by monitoring both lipid hydroperoxide and headspace hexanal formation. The degree of positive charge on the protein-stabilized emulsion droplets was not the only factor involved in the inhibition of lipid oxidation because the charge of the emulsion droplets (WPI > casein > or = SPI) did not parallel oxidative stability. Other potential reasons for differences in oxidative stability of the protein-stabilized emulsions include differences in interfacial film thickness, protein chelating properties, and differences in free radical scavenging amino acids. This research shows that differences can be seen in the oxidative stability of protein-stabilized emulsions; however, further research is needed to determine the mechanisms for these differences.     

Effects of sugars on whey protein isolate gelation

Whey protein isolate (WPI) gels were prepared from solutions containing ribose or lactose at pH values ranging from 6 to 9. The gels with added lactose had no color development, whereas the gels with added ribose were orange/brown. Lactose stabilized the WPI to denaturation, which increased the time and temperature required for gelation, thus decreasing the fracture modulus of the gel compared to the gels with added ribose and the gels with no sugar added. Ribose, however, favored the Maillard reaction and covalent cross-linking of proteins, which increased gel fracture modulus. The decreased pH caused by the Maillard reaction in the gels containing ribose occurred after protein denaturation and gelation, thus having little if any effect on the gelation process.     

超高压处理对杏汁香气成分的影响

为了探讨超高压处理对杏汁香气成分的影响,将杏原汁在500 MPa压力、25℃温度条件下处理20 min后,经顶空固相微萃取与毛细管气相色谱-质谱联用技术检测超高压处理前后杏汁香气成分的变化,并用面积归-化法测定了各种成分的质量分数.结果表明超高压处理后对杏汁中香气成分的有较明显影响,其中己醛、2-己烯醛、糠醛、己醇、叶醇、芳樟醇、橙花醇、β-苯乙醇等香味成分的质量分数分别增长了68.14%、95.26%、46.76%、61.11%、58.56%、35.75%、37.75%和42.30%;酯类、内酯类的香气成分的含量有所降低;酮类香味成分的含量则没有明显变化.感官评定表明:超高压处理不仅能很好地保持了杏的特征香气,而且使杏汁的青鲜香气更加突出,有利于产品风味品质的提高,这与杏汁中香气成分检测结果相一致,因此对杏加工来说超高压处理是一种很有前景的冷加工技术.     

不同热处理大豆分离蛋白凝胶冻藏特性

为探究冻藏过程中不同加热温度处理大豆分离蛋白(soybean isolate protein,SPI)凝胶特性变化及评估不同热处理对SPI凝胶冻藏特性的影响。该文以65、90和135℃3个不同温度处理所得SPI为研究对象(分别记为65SPI、90SPI和USPI),采用离心法、质构分析法、可溶蛋白含量测定和电泳等方法对其冻藏过程中的凝胶持水性、凝胶硬度、凝胶弹性、可溶蛋白含量及亚基组成和凝胶作用力进行了分析研究。结果表明:随冻藏时间延长,不同温度处理SPI凝胶持水性、凝胶弹性和凝胶可溶蛋白含量呈下降趋势,而凝胶硬度呈增大趋势。凝胶持水性、弹性的下降和凝胶硬度的升高标志着凝胶品质的劣变。不同温度处理对SPI凝胶的冻前凝胶特性和冻藏特性有较大影响,65和90℃的温度处理降低了冻前SPI凝胶的持水性,增强了冻前SPI凝胶硬度,有更多的β和B亚基参与了凝胶形成,冻藏前后的亚基组成没有变化;超高温瞬时加热(ultra high temperature,UHT)处理则降低了冻前SPI凝胶硬度,冻藏过程中可溶蛋白含量大幅下降且可溶蛋白中β和B亚基含量下降。3种温度处理SPI的凝胶劣变程度均高于未处理SPI。加热处理会造成SPI发生部分或完全变性,变性后疏水基团的暴露会加快蛋白凝胶形成过程中聚集速率,进而增大粗糙凝胶结构形成的几率,而粗糙凝胶网络在冻藏过程中其劣变程度更甚于未加热SPI。由此可知,加热处理尽管在一定程度上增大了凝胶硬度,但会加速其凝胶品质冻藏劣变。     

Enzymatic hydrolysis of heat-induced aggregates of whey protein isolate

The effects of heat-induced denaturation and subsequent aggregation of whey protein isolate (WPI) solutions on the rate of enzymatic hydrolysis was investigated. Both heated (60 °C, 15 min; 65 °C, 5 and 15 min; 70 °C, 5 and 15 min, 75 °C, 5 and 15 min; 80 °C, 10 min) and unheated WPI solutions (100 g L(-1) protein) were incubated with a commercial proteolytic enzyme preparation, Corolase PP, until they reached a target degree of hydrolysis (DH) of 5%. WPI solutions on heating were characterized by large aggregate formation, higher viscosity, and surface hydrophobicity and hydrolyzed more rapidly (P < 0.001) than the unheated. The whey proteins exhibited differences in their susceptibility to hydrolysis. Both viscosity and surface hydrophobicity along with insolubility declined as hydrolysis progressed. However, microstructural changes observed by light and confocal laser scanning microscopy (CLSM) provided insights to suggest that aggregate size and porosity may be complementary to denaturation in promoting faster enzymatic hydrolysis. This could be clearly observed in the course of aggregate disintegration, gel network breakdown, and improved solution clarification.     

Rheological and physical properties of derivitized whey protein isolate powders

Pregelatinized starch is employed in many food applications due to the instantaneous nature of thickening and stability imparted by modification. Proteins, however, have been excluded as a viscosifying agent due to requisite thermal treatments required to create structure. Whey protein isolate gels were produced while manipulating heating time, pH, and mineral type/content, producing a variety of gel types/networks. Gels were frozen, freeze-dried, and ground into a powder. Once reconstituted in deionized water, gel powders were evaluated based on solubility studies, rotational viscometry, and electrophoresis. The protein powder exhibiting the largest apparent viscosity, highest degree of hydrolysis, and greatest solubility was selected for pH and temperature stability analyses and small amplitude oscillatory rheology. This processing technique manipulates WPI into a product capable of forming cold-set weak gel structures suitable for thickening over a wide range of temperature and pH food systems.     

高压均质-冷冻干燥技术制备大豆分离蛋白微粒及其功能特性

为了进一步改善大豆分离蛋白的分散性及功能性质,该研究以大豆分离蛋白为原料,通过对天然大豆分离蛋白进行高压高剪切处理并联合冷冻干燥技术,制备大豆分离蛋白微粒,考察压力(60～100 MPa)对大豆分离蛋白微粒尺寸、功能性质及结构特性的影响,探究其构效关系.结果表明:随着压力逐渐增加,大豆分离蛋白平均粒径大幅度减小,粒径分...     

Effect of additives on subcritical water hydrolysis of whey protein isolate

The objective was to examine the effect of the additives acetic acid, lactic acid, sodium bicarbonate, sodium chloride, and sodium hydroxide on the hydrolysis of whey protein isolate with subcritical water. A screening experimental design was used to study the effect of temperature, time, and additives. The most influential additive, sodium bicarbonate, along with temperature and time was used in a second experimental design to predict the treatment conditions to maximize the degree of hydrolysis and production of free amino acids. The maximum degree of hydrolysis achieved was 50% at a concentration of 1.24 M sodium bicarbonate, 291 °C, and 28 min. The highest concentration of total amino acids was 83.0 mg/g of whey protein isolate with 0.83 M sodium bicarbonate at 264 °C for 29 min. Compared to water alone, sodium bicarbonate increased the degree of hydrolysis 4-fold and the production of amino acids by 44% and decreased peptides' molecular weight.     

Charge modifications to improve the emulsifying properties of whey protein isolate

Whey protein isolate was modified by ethylene diamine in order to shift its isoelectric point to an alkaline pH. The extent of the modification was studied using SDS-PAGE and MALDI-TOF mass spectrometry. The modified whey proteins were used as an emulsifier to stabilize oil-in-water emulsions at acidic and neutral pH ranges, and their emulsifying properties were compared with that of the unmodified whey proteins and with the previously studied ethylene diamine modified sodium caseinate. The emulsifying activity of the modified whey proteins was similar to that of the unmodified ones, but the stability of an emulsion at pH 5 was significantly improved after the modification. Charge and coverage of droplet surface and the displacement of the interfacial proteins by surfactant Tween 20 were further studied as a function of pH. As compared with the unmodified whey proteins, the modified ones were proven to cover the interface more efficiently with extensive surface charge at pH 5, although the interfacial layer was less resistant to the surfactant displacement.     

Inverse gas chromatographic method for measurement of interactions between soy protein isolate and selected flavor compounds under controlled relative humidity

An inverse gas chromatographic (IGC) method was developed to study the binding interactions between selected volatile flavor compounds and soy protein isolate (SPI) under controlled relative humidity (RH). Three volatile probes (hexane, 1-hexanol, and hexanal) at very low levels were used to evaluate and validate system performance. On the basis of the thermodynamic data and the isotherms measured at 0% RH, 1-hexanol and hexanal had higher binding affinities than hexane, which could be attributed to hydrogen-bonding interactions with SPI. At 30% RH, 1-hexanol and hexanal were retained less than at 0% RH, indicating possible competition for binding sites on the SPI surface between water and volatile probe molecules. Results showed that the thermodynamic data determined were comparable to the available literature values. Use of IGC allowed for the rapid and precise generation of sorption isotherms. Repeatability between replicate injections and reproducibility across columns were very good. IGC is a potentially high-throughput method for the sensitive, precise, and accurate measurement of flavor-ingredient interactions in low-moisture food systems.     

高压处理对牛乳清蛋白体外消化性的影响

为了评价牛乳清蛋白(WPI)加压凝胶的消化性,使用未加压WPI及加压(200～600MPa)WPI凝胶进行了体外模拟胃蛋白酶-胰蛋白酶消化率的测定,并用十二烷基磺酸钠-聚丙烯酰胺(SDS-PAGE)电泳的方法分析了不同压力对WPI构成蛋白消化性的影响。结果表明,经过胃蛋白酶和胰蛋白酶的消化后,加压WPI凝胶消化率显著高于未加压WPI,分别为72.59%和61.02%;400MPa以上加压处理后,加压WPI凝胶消化率未见有明显的变化(p<0.05)。尽管未加压WPI中β-乳球蛋白不易被胃蛋白酶消化,400MPa加压WPI凝胶中的β-乳球蛋白则能被胃蛋白酶部分消化,而600MPa加压后β-乳球蛋白几乎被完全消化。此外,在胃蛋白酶消化阶段,有3500～6500u范围的多肽产物生成。     

Isothermal calorimetry study of calcium caseinate and whey protein isolate edible films cross-linked by heating and gamma-irradiation

The contribution of thermal and radiative treatments as well as the presence of some excipients, namely glycerol, carboxymethylcellulose (CMC), pectin, and agar, on the formation of protein-protein interactions as well as the formation and loss of protein-water interactions was investigated by means of differential scanning calorimetry in an isothermal mode. Protein-water interactions were assessed through measurement of the heat of the wetting parameter. Isothermal calorimetry measurements pointed out that gamma-irradiation does not favor protein-water interactions, as reflected by its endothermic contribution (P < or = 0.05) to the heat of wetting values. Although significant (P < or = 0.05), the effect of the thermal treatment on endothermic responses using isothermal calorimetry was found to be somewhat lower. Among excipients added to biofilm formulations, glycerol generated the most important losses of protein-water interactions, as inferred by its significant (P < or = 0.05) endothermic impact on the heat of wetting values.     

Formation of disulfide bonds in acid-induced gels of preheated whey protein isolate

Cold gelation of whey proteins is a two-step process. First, protein aggregates are prepared by a heat treatment of a solution of native proteins in the absence of salt. Second, after cooling of the solution, gelation is induced by lowering the pH at ambient temperature. To demonstrate the additional formation of disulfide bonds during this second step, gelation of whey protein aggregates with and without a thiol-blocking treatment was studied. Modification of reactive thiols on the surface of the aggregates was carried out after the heat-treatment step. To exclude specific effects of the agent itself, different thiol-blocking agents were used. Dynamic light scattering and SDS-agarose gel electrophoresis were used to show that the size of the aggregates was not changed by this modification. The kinetics of gelation as determined by the development of pH and turbidity within the first 8 h of acidification were not affected by blocking thiol groups. During gelation, formation of large, covalently linked, aggregates occurred only in the case of unblocked WPI aggregates, which demonstrates that additional disulfide bonds were formed. Results of permeability and confocal scanning laser microscope measurements did not reveal any differences in the microstructure of networks prepared from treated or untreated whey protein aggregates. However, gel hardness was decreased 10-fold in gels prepared from blocked aggregates. Mixing different amounts of blocked and unblocked aggregates allowed gel hardness to be controlled. It is proposed that the initial microstructure of the gels is primarily determined by the acid-induced noncovalent interactions. The additional covalent disulfide bonds formed during gelation are involved in stabilizing the network and increase gel strength.     

Binding between bixin and whey protein at pH 7.4 studied by spectroscopy and isothermal titration calorimetry

Bixin is the major coloring component of annatto used in manufacturing colored cheeses, but its presence in liquid whey causes undesirable quality of the recovered whey protein ingredients. The objective of this work was to study molecular binding between bixin and three major whey proteins (β-lactoglobulin, α-lactalbumin, and bovine serum albumin) at pH 7.4 using UV-vis absorption spectroscopy, fluorescence spectroscopy, isothermal titration calorimetry, and circular dichroism. These complementary techniques illustrated that the binding is a spontaneous complexation process mainly driven by hydrophobic interactions. The complexation is favored at a lower temperature and a higher ionic strength. At a lower temperature, the binding is entropy-driven, while it changes to an enthalpy-driven process at higher temperatures. The binding also increases the percentage of unordered secondary structures of proteins. Findings from this work can be used to develop whey protein recovery processes for minimizing residual annatto content in whey protein ingredients.     

Formation of whey protein isolate (WPI)-dextran conjugates in aqueous solutions

The conjugation reaction between whey protein isolate (WPI) and dextran in aqueous solutions via the initial stage of the Maillard reaction was studied. The covalent attachment of dextran to WPI was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with both protein and carbohydrate staining. The formation of WPI-dextran conjugates was monitored by a maximum absorbance peak at approximately 304 nm using difference UV spectroscopy. The impact of various processing conditions on the formation of WPI-dextran conjugates was investigated. The conjugation reaction was promoted by raising the temperature from 40 to 60 degrees C, the WPI concentration from 2.5 to 10%, and the dextran concentration from 10 to 30% and lowering the pH from 8.5 to 6.5. The optimal conjugation conditions chosen from the experiments were 10% WPI-30% dextran and pH 6.5 at 60 degrees C for 24 h. WPI-dextran conjugates were stable under the conditions studied.     

Effects of ionic strength on the enzymatic hydrolysis of diluted and concentrated whey protein isolate

To identify the parameters that affect enzymatic hydrolysis at high substrate concentrations, whey protein isolate (1-30% w/v) was hydrolyzed by Alcalase and Neutrase at constant enzyme-to-substrate ratio. No changes were observed in the solubility and the aggregation state of the proteins. With increasing concentration, both the hydrolysis rate and the final DH decreased, from 0.14 to 0.015 s(-1) and from 24 to 15%, respectively. The presence of 0.5 M NaCl decreased the rate of hydrolysis for low concentrations (to 0.018 s(-1) for 1% WPI), resulting in similar rates of hydrolysis for all substrate concentrations. The conductivity increase (by increasing the protein concentration, or by addition of NaCl) has significant effects on the hydrolysis kinetics, but the reason for this is not yet well understood. The results show the importance of conductivity as a factor that influences the kinetics of the hydrolysis, as well as the composition of the hydrolysates.     

Adsorption of whey protein isolate at the oil-water interface as a function of processing conditions: a rheokinetic study

In this paper we present surface dynamic properties (interfacial tension and surface dilational properties) of a whey protein isolate with a high content of beta-lactoglobulin (WPI) adsorbed on the oil-water interface as a function of adsorption time. The experiments were performed at constant temperature (20 degrees C), pH (5), and ionic strength (0.05 M). The surface rheological parameters and the interfacial tension were measured as a function of WPI concentration (ranging from 1 x 10(-)(1) to 1 x 10(-)(5)% w/w) and different processing factors (effect of convection and heat treatment). We found that the interfacial pressure, pi, and surface dilational modulus, E, increase and the phase angle, phi, decreases with time, theta, which should be associated with WPI adsorption. These phenomena have been related to diffusion of the protein toward the interface (at short adsorption time) and to the protein unfolding and/or protein-protein interactions (at long-term adsorption) as a function of protein concentration in solution and processing conditions.     

Post-harvest disorders and mineral composition of apple fruits as affected by pre-harvest calcium treatments

Abstract

The effect of pre-harvest calcium sprays, CaCl₂ and Ca (NO₃)_2, on apple (Malus domestica) N, P, K, Ca and Mg content, storability and incidence of physiological disorders (superficial scald, bitter pit, physiological spot) was studied using two cultivars, ‘Krameri Tuviõun’ and ‘Talvenauding’. In the first year calcium treatment did not reduce any physiological disorders or loss of marketable yield. In the second year calcium treatment reduced storage losses until four months after storage. Bitter pit in ‘Krameri Tuviõun’ was unaffected by calcium treatment, but physiological spot of ‘Krameri Tuviõun’ and superficial scald of ‘Talvenauding’ were reduced by calcium treatment in the second year. Both physiological disorders of 'Krameri Tuviõun’ correlated negatively with Mg and P content and Mg/Ca ratio in apples. Superficial scald of ‘Talvenauding’ correlated negatively with Ca content and positively with K/Ca ratio, N/Ca ratio and Mg/Ca ratio in fruits. The conclusion can be made that content of Ca and its ratios with other nutrients plays an important role in the development of superficial scald on ‘Talvenauding’ fruits, and increasing Ca content in fruit can therefore be recommended for improving post-harvest quality of this cultivar. Since bitter pit in 'Krameri Tuviõun’ was not reduced by calcium treatment, it would be worth trying other measures for this cultivar.