Properties of whey protein isolates extruded under acidic and alkaline conditions

Whey proteins have wide acceptance and use in many products due to their beneficial nutritional properties. To further increase the amount of whey protein isolates (WPI) that may be added to products such as extruded snacks and meats, texturization of WPI is necessary. Texturization changes the folding of globular proteins to improve interaction with other ingredients and create new functional ingredients. In this study, WPI pastes (60% solids) were extruded in a twin-screw extruder at 100 degrees C with 4 pH-adjusted water streams: acidic (pH 2.0 +/- 0.2) and alkaline (pH 12.4 +/- 0.4) streams from 2 N HCl and 2 N NaOH, respectively, and acidic (pH 2.5 +/- 0.2) and alkaline (pH 11.5 +/- 0.4) electrolyzed water streams; these were compared with WPI extruded with deionized water. The effects of water acidity on WPI solubility at pH 7, color, microstructure, Rapid Visco Analyzer pasting properties, and physical structure were determined. Alkaline conditions increased insolubility caused yellowing and increased pasting properties significantly. Acidic conditions increased solubility and decreased WPI pasting properties. Subtle structural changes occurred under acidic conditions, but were more pronounced under alkaline conditions. Overall, alkaline conditions increased denaturation in the extruded WPI resulting in stringy texturized WPI products, which could be used in meat applications.     

Encapsulation of eugenol using Maillard-type conjugates to form transparent and heat stable nanoscale dispersions

Maillard-type protein-carbohydrate conjugates are known for their excellent emulsifying properties and have been used to encapsulate volatile oils and flavor compounds. In the present study, eugenol was used as a model compound for encapsulation in conjugates of whey protein isolate (WPI) and maltodextrins (MD) made using different WPI:MD mass ratios and MD chain lengths. The encapsulation involved two steps, emulsifying an oil phase of eugenol dissolved in hexane into an aqueous phase with dissolved conjugates and spray drying the emulsion. Mass yield up to 82.7 g/100 g and encapsulation efficiency as high as 35.7 g/100 g eugenol were observed. After hydrating spray-dried powders, several samples with an eugenol content above its solubility limit demonstrated transparent dispersions at pH 3.0 and 7.0 after heating at 80 °C for 15 min, corresponding to mean diameters smaller than ca. 100 nm. One treatment also showed transparent dispersions after heating at pH 5.0, which is near the isoelectric point of whey proteins, in contrast to gel formation for a control prepared with a mixture of non-conjugated WPI and MD. The present study demonstrates potential to produce food grade nanoscale systems for delivering lipophilic bioactives in functional beverages, without adversely affecting their visual appearance.     

Role of Covalent and Noncovalent Interactions in the Formation of Films from Unheated Whey Protein Solutions Following pH Adjustment

ABSTRACT: Films were formed from heated whey protein isolate (WPI) solutions (heated [H] films) and from unheated WPI solutions following adjustment to pH 11, with subsequent readjustment to pH 7 (unheated, readjusted [UR] films) or without readjustment to pH 7 (unheated, unadjusted films [UU] films). UU and UR films had significantly lower % elongation, tensile strength, and Young's modulus than H films. Film solubility and dispersion in water were in the order: H films < UU films < UR films. Free sulphydryl groups were lower and disulphide-mediated polymerization was higher in heated than in unheated WPI solutions whereas solubility of H films increased in the presence of dithiothreitol.     

Iron‐encapsulated cold‐set whey protein isolate gel powder – Part 1: Optimisation of preparation conditions and in vitro evaluation

A central composite design with a quadratic model was used to investigate the effects of three independent variables involved in the synthesis of iron‐encapsulated cold‐set whey protein isolate gel (WPI) on encapsulation efficiency (EE) and L*, a*, b* colour characteristics. The optimal conditions for maximum EE with minimum colour alteration were determined as 6.8% WPI, 18.8 mM iron and pH 7. In an in vitro gastrointestinal assay, only about 28% of the encapsulated iron was released in the gastric condition (with pepsin at pH 1.2), compared to 95% in the intestinal condition (with pancreatin at pH 7.5).     

槲皮素及其乙酸乙酯衍生物的平衡溶解度和油水分配系数的测定

通过测定槲皮素及其酯化物的平衡溶解度和油水分配系数,检测酯化后的槲皮素衍生物脂溶性是否提高。采用高效液相色谱法测定槲皮素及3,7,3’,4’-O-四乙酸乙酯槲皮素在水、不同缓冲盐溶液中的质量浓度;采用摇瓶-液相色谱法,测定二者在正辛醇-水、缓冲液体系中的油水分配系数。37 ℃时,槲皮素在水中的平衡溶解度为23.02 μg/mL,而3,7,3’,4’-O-四乙酸乙酯槲皮素在水中的平衡溶解度更小,为10.23 μg/mL。随着溶剂pH值的增大,槲皮素的平衡溶解度增大,3,7,3’,4’-O-四乙酸乙酯槲皮素变化并不显著。槲皮素、3,7,3’,4’-O-四乙酸乙酯槲皮素在正辛醇-水体系条件下的油水分配系数分别为1.819、3.696。酯化修饰后的3,7,3’,4’-O-四乙酸乙酯槲皮素脂溶性高于槲皮素。     

乳清分离蛋白-植物甾醇纳米颗粒的制备与表征

本研究旨在使用乳清分离蛋白(Whey protein isolate, WPI)和植物甾醇(Phytosterols, PSs)采用超声辅助反溶剂沉淀法制备不同质量比的乳清分离蛋白-植物甾醇(WPSs)纳米颗粒,采用动态光散射技术、扫描电子显微镜、傅里叶红外光谱等技术对样品的形貌和结构进行表征,并对样品的pH稳定性、盐稳定性及体外释放进行了研究。结果表明：WPI/PSs质量比从50:1降低到10:1时,WPSs纳米颗粒粒径减小(252.77 ~ 215.90 nm),颗粒表面Zeta负电位增加(-31.27 ~ -37.37 mV),PSs包封率降低(95.39 ~ 81.55%)；差示扫描量热结果表明PSs成功包埋在WPI中；红外光谱分析表明,PSs改变了WPI的二级结构；微观结构显示,随着PSs浓度的增加,WPSs纳米颗粒逐渐从清晰的球形变成网络结构、块状结构。另外,WPI/PSs质量比低于25:2时,WPSs纳米颗粒的复溶性较差。研究还发现,这些颗粒在高浓度的盐环境中以及在模拟胃条件下具有良好的稳定性。该研究证实,WPI包埋PSs后,其结构和形貌发生了改变,并且有利于小肠对PSs的吸收。     

Solubility and mechanical properties of heat-cured whey protein-based edible films compared with that of collagen and natural casings

Water solubility, tensile strength (TS), wet strength (WS) and elongation at break (%E) of whey protein isolate (WPI) films were compared to that of collagen films and natural casings. Increase in heat-curing temperature and time caused decreased ( P <  0.001) water solubility and increased TS and WS of the films. Heat-cured WPI films with similar properties (solubility, TS, WS and %E) to collagen films were obtained by optimizing heat-curing conditions. Overall, natural casings had lower solubility, TS and %E but higher WS than collagen and heat-cured WPI films. Heat-cured WPI films have the potential as an alternative to collagen films and casings.     

Synergistic effects of whey protein–polysaccharide complexes on the controlled release of lipid‐soluble and water‐soluble vitamins in W1/O/W2 double emulsion systems

Lipid‐soluble vitamin E (V_E) and water‐soluble vitamin B₂ (V_B2) can be encapsulated together in the same water–oil–water emulsion system. The ability of whey protein isolate (WPI)–polysaccharide complexes to synergistically control the release rates in such a system was investigated. The complexes studied were WPI–low methoxyl pectin (LMP) and WPI–κ‐carrageenan (KCG). The encapsulation efficiency of V_E and V_B2 of the WPI system is 66% and 64% and increases by about 1.4‐ and 1.2‐fold in the WPI–LMP and WPI–KCG complexes, respectively, which serve as selective non‐pH‐dependent barriers against enzyme attack. These complexes also greatly ameliorate the controlled release rates of both vitamins. The low‐charge LMP, with its multiple protein‐ and oil‐binding abilities, exhibits greater synergistic effects than the high‐charge KCG.     

Preparation and characterization of kafirin‐quercetin film for packaging cod during cold storage

This study mainly evaluated the physical properties of kafirin‐quercetin (KQ) edible films and their application on the quality of cod fillets during cold storage. The results showed that the addition of quercetin significantly increased mechanical properties of KQ films, while decreased water vapor permeability, water solubility, and transparency. As quercetin was 0.4% (wt/vol), the film had the highest tensile strength (4.96 ± 1.23 MPa), the lowest water vapor permeability (1.08 ± 0.09 g·mm·m^?2·h·KPa^?1) and water solubility (22.02 ± 0.45%). Moreover, compared with the pure kafirin and polythylene films, KQ films could effectively inhibit the cod meat deterioration by restraining the growth of microorganisms and decreasing TVB‐N and TBARs. The KQ_0.4% film was the best to prolong the shelf life of cod fillets during cold storage. Therefore, KQ edible films could be used as a potential food packaging material to protect and retain the quality of aquatic products.     

雨生红球藻虾青素纳米颗粒的制备及稳定性研究

为改善雨生红球藻虾青素的稳定性和水溶性,本文使用蜗牛酶进行雨生红球藻虾青素的破壁提取,并以阿拉伯胶和乳清蛋白粉（富含乳脂肪球膜）为壁材,利用复合凝聚法制备虾青素纳米颗粒,此外对纳米颗粒的稳定性进行研究。结果表明:虾青素纳米颗粒的最佳制备条件为:pH4.0,乳清蛋白与阿拉伯胶质量比为2:1,虾青素浓度为60 μmol/L,此工艺条件下虾青素的包封率为92.93%±0.19%。该纳米颗粒平均粒径为265.71±0.55 nm,Zeta电位为?13.44±0.14 mV,并具备良好的贮藏稳定性,在4 ℃条件下贮藏15 d,粒径增幅仅为6.1%,虾青素保留率为90.78%±0.25%,DPPH清除率为79.31%±0.18%。本研究改善了雨生红球藻虾青素的稳定性和水溶性,为虾青素的高效利用提供了技术支持。     

Gelling properties of whey protein isolate: influence of calcium removal by dialysis or diafiltration at acid or neutral pH

Dialysis of whey protein isolates (WPI) removed much more calcium when carried out at an acid pH (close to 4.0) than at neutral pH. Diafiltration at acid pH was also effective. The characteristics of thermally-induced gels prepared from WPI dialysed at acid or neutral pH were studied at pH 3.75 or pH 7.0, respectively, and at calcium concentrations ranging from 0 to about 60mM (with addition of calcium chloride). The water-holding capacity (WHC) and elasticity of gels increased with decreasing calcium concentration, at both pHs. Gel firmness was maximum at 10–20 mM calcium. The solubility of the protein constituents of WPI gels in a pH 8.0 buffer was high in the case of acid gels (especially at calcium concentrations lower or equal to 20 mM) and low for neutral gels at all calcium concentrations. Protein solubility values in the presence or absence of denaturing and reducing agents reflect the existence of intermolecular disulphide bonds in neutral gels and their absence in acid gels.     

Enhancement of the functional properties of whey protein by conjugation with maltodextrin under dry‐heating conditions

Conjugation of whey protein isolate (WPI) and maltodextrin (MD, dextrose equivalent of 6) was achieved by dry‐heating at an initial pH of 7.0, at 60 °C and 79% relative humidity, with WPI: MD6 ratio of 1:1, for up to 24 h. Conjugation was achieved with limited development of colour and advanced Maillard products on 24 h of heating. Conjugation increased the protein solubility at pH 4.5, by 7.1–8.5%, compared to the unheated and heated WPI controls. Conjugation of WPI with MD6 enhanced the stability and retention of clarity in protein solutions heated at 85 °C for 10 min with 50 mM added NaCl.     

Water Vapor Permeability and Solubility of Films from Hydrolyzed Whey Protein

ABSTRACT: The effects of whey protein hydrolysis on film water vapor permeability (WVP) and solubility at 3 plasticizer levels were studied. Little or no significant difference (p > 0.05) appeared for film WVP between unhydrolyzed whey protein isolate (WPI), 5.5% degree of hydrolysis (DH) WPI and 10% DH WPI films at comparable plasticizer contents. However, increase in glycerol (gly) content significantly increased film WVP. Thus, reduction in WPI molecular weight (MW) through hydrolysis may be a better approach to improving film flexibility than addition of plasticizer. Both 5.5% and 10% DH WPI had significantly different (p ≤ 0.05) film solubility compared to unhydrolyzed WPI. Soluble Protein (SP) and total soluble matter (TSM) of hydrolyzed WPI films were much higher than for unhydrolyzed WPI films.     

Synthesis of Quercetin‐3‐O‐Glucoside from Rutin by Penicillium decumbens Naringinase

Enzymatic bioconversion of rutin to quercetin‐3‐O‐glucoside (Q‐3‐G) by Penicillium decumbens naringinase was increased with reaction pH increased approximately to pH 6.0. It resulted in greater than 92% production of Q‐3‐G due to the removal of the terminal rhamnose at the controlled pH 6.0. The enzymatic bioconversion of rutin to Q‐3‐G was repetitively performed, yielding 84% after 5 batches with little quercetin formation. Interestingly, the water solubility of Q‐3‐G was enhanced 69‐ and 328‐fold over those of rutin and quercetin, which may make Q‐3‐G more bioavailable in food. Q‐3‐G was approximately 6‐ and 1.4‐fold more potent than rutin as an inhibitor of human intestinal maltase and human DL‐3‐hydroxy‐3‐methylglutalyl coenzyme A reductase. Q‐3‐G was less potent (16‐ and 1.3‐fold, respectively) than quercetin as an inhibitor of these enzymes. However, the results suggest that Q‐3‐G may be confirmed more effective and bioavailable food component than rutin and even quercetin because of its enhanced solubility and inhibitory properties. Practical Application : Bioconverted intermediate, quercetin‐3‐O‐glucoside (Q‐3‐G), was found and confirmed to be largely more soluble than rutin and quercetin in water solution, which might make it more bioavailable as food ingredient. In addition, Q‐3‐G inhibited mildly the intestinal maltase, which might act as antidiabetic substance by modulating the adsorption of glucose in the intestine.     

Effect of microbial transglutaminase treatment on thermal stability and pH-solubility of heat-shocked whey protein isolate

Whey protein isolate (WPI) dispersions (5% protein, pH 7.0) were subjected to heat-shock at 70 °C for 1, 5 and 10 min. The heat-shocked WPI dispersions were treated with microbial transglutaminase (MTGase) enzyme, and thermal properties and pH-solubility of the treated proteins were investigated. Heat-shocking of WPI for 10 min at 70 °C increased the thermal denaturation temperature (T_d) of β-lactoglobulin in WPI by about 1.5 °C. MTGase treatment (30 h, 37 °C) of the heat-shocked WPI significantly increased the T_d of β-lactoglobulin by about 6.3–7.3 °C when compared with heat-shocked only WPI at pH 7.0. The T_d increased by about 13–15 °C following pH adjustment to 2.5; however, the T_d of heat-shocked WPI was not substantially different from heat-shocked and MTGase-treated WPI at pH 2.5. Both the heat-shocked and the heat-shocked-MTGase-treated WPI exhibited U-shaped pH-solubility profiles with minimum solubility at pH 4.0–5.0. However, the extent of precipitation of MTGase-treated WPI samples at pH 4.0–5.0 was much greater than all heat-shocked and native WPI samples. The study revealed that while MTGase cross-linking significantly enhanced the thermal stability of β-lactoglobulin in heat-shocked WPI, it caused pronounced precipitation at pH 4.0–5.0 via decreasing the hydrophilic/hydrophobic ratio of the water-accessible protein surface.     

Microcoagulation of a Whey Protein Isolate by Extrusion Cooking at Acid pH

A whey protein isolate (WPI) was coagulated by thermomechanical processing in a twin screw extruder. Nonaggregated semi-solid spreads were obtained only in the pH range 3.5–3.9, at ca 20% protein (77% water), a barrel temperature of 90–100°C and a screw speed of 100–200 rpm. WPI extrusion-coagulated at pH 3.9 displayed a high nitrogen solubility (NSI) (43–47%). Electrophoresis indicated that the β-lactoglobulin constituent was entirely soluble in 1% SDS, while scanning calorimetry revealed about 82% protein unfolding. WPI extrusion-coagulated at pH 4.5–6.8 displayed lower NSI (25%), were less soluble in 1% SDS, were 88% unfolded and had grainy texture. Light microscopy, centrifugation in glycerol solutions, and laser diffractometry indicated the acid spread (pH 3.9) was composed of small coagulated particles, mean diameter 11.5 μm (volume basis).     

Effect of pH and addition of corn oil on the properties of whey protein isolate-based films using response surface methodology

Response surface methodology (RSM) was used to investigate pH and corn oil (CO) effects on the properties of films formed from whey protein isolate (WPI). Test films were evaluated for tensile strength (TS), puncture strength (PT), percentage elongation at break point (E), water vapour permeability (WVP) and oxygen permeability (OP). TS of WPI films increased with increasing pH, while addition of CO produced no trend. However, when WPI solution pH increased >10.0, film TS generally decreased with CO addition (>11%). E values increased dramatically with increasing levels of CO when pH for WPI solutions were >8.5. However, pH had no effect on E values. WPI solutions possessing high pH values (maximum pH value of 10.62) produced WPI films with the highest PT values. WVP had a quadratic relationship with pH and CO addition. OP had an inversely linear relationship with increasing pH (6.5–10.5) and a quadratic relationship with CO addition. Optimal pH (9.88) and CO level (2.93%), determined from physical test film data, were predicted by RSM.     

Improvement of functional properties of whey protein isolate through glycation and phosphorylation by dry heating

Whey protein isolate (WPI) was glycated with maltopentaose (MP) through the Maillard reaction, and the MP-conjugated WPI (MP-WPI) was then phosphorylated by dry heating in the presence of pyrophosphate. Glycation occurred efficiently, and the sugar content of WPI increased approximately 19.9% through the Maillard reaction. The phosphorylation of MP-WPI was enhanced with an increase in the dry-heating time from 1 to 5 d, and the phosphorus content of WPI increased approximately 1.05% by dry heating at pH 4.0 and 85°C for 5 d in the presence of pyrophosphate. The electrophoretic mobility of WPI increased with an increase in the phosphorylation level. The stability of WPI against heat-induced insolubility at pH 7.0 was improved by conjugation with MP alone, and further improved by phosphorylation. Although the emulsifying activity of WPI was barely affected by glycation and phosphorylation, the emulsifying stability of phosphorylated MP-WPI (5 d), was 2.2 times higher than that of MP-WPI. Gelling properties such as hardness, resiliency, and water-holding capacity of heat-induced WPI gel were markedly improved, and the gel was rendered transparent by phosphorylation. The calcium phosphate-solubilizing ability of WPI was enhanced by phosphorylation. These results suggested that phosphorylation by dry heating in the presence of pyrophosphate after conjugation with MP is a useful method for improving the functional properties of WPI.     

Development and characterisation of whey protein micro-beads as potential matrices for probiotic protection

This study evaluated the efficacy of whey protein isolate (WPI) as an encapsulation matrix for the maintenance of Lactobacillus rhamnosus GG viability during simulated gastro-intestinal studies. Micro-bead characteristics were investigated using microscopy, chromatography, laser diffractometry and zeta potential analysis. Heat-treated WPI (11%, w/v) blended with stationary phase cultures demonstrated an instant gelation impetus in acetate buffer (0.5 M), tempered to 35 °C in the presence of Tween-20 (0.04%). Atomic force microscopy (AFM) demonstrated that micro-bead extrusion at pH 4.6 fuelled strong cohesive interactions within protein-probiotic amalgams; an electrostatic alliance further highlighted by zeta potential analysis. Optimization of encapsulation conditions generated self-supporting structures (200 ± 1.2 μm) with high micro-bead strength and individual loading capacity of 2.7 × 10⁴ cfu/micro-bead. Plate enumeration demonstrated that micro-bead extrusion had no detrimental effect on cell viability due to the perpetuation of stationary phase concentrations (10⁹ cfu/mL). This finding was further validated by LIVE/DEAD microscopy staining, which visualized the homogenous distribution of live probiotics throughout micro-bead matrices. Following 3 h in vitro stomach incubation (pH 1.8; 37 °C), micro-beads laden with 10¹⁰ cfu demonstrated acid-stability and peptic-resistance, characteristics required for optimum probiotic refuge. However, enzyme-activated intestinal conditions catalysed a synergistic response engaging rapid matrix disintegration and controlled probiotic release. Matrix digestion was monitored by chromatography, which witnessed the sequential release of peptides <2 kDa after 30 min. In conclusion, this study led to the development and design of a protein encapsulation polymer based on congruent matrix interactions for reinforced probiotic protection during challenging situations for their targeted delivery to intestinal absorption sites.     

Partition and stability of resveratrol in whey protein isolate oil-in-water emulsion: Impact of protein and calcium concentrations

Whey protein isolate (WPI) is often used in food emulsions and can also interact with resveratrol, a natural amphiphilic polyphenol, this interaction being improved by heat-denaturation. In this study, oil-in-water emulsions stabilised by heat-denatured WPI in the absence and presence of CaCl₂ were characterised in terms of size, ζ-potential and protein partition. Partition and stability of resveratrol were also studied as a function of WPI and calcium concentrations. Size of WPI emulsions was dependent on the protein content at the oil–water interface. Partition of resveratrol and WPI was positively proportional at the oil–water interface and in the continuous phase. The stability of resveratrol increased as the concentration of WPI increased, but decreased when the concentration of calcium exceeded 0.20 mm. These data should be useful for simultaneous encapsulation of hydrophobic and amphiphilic bioactive components in a single emulsion and the protection of the inner oil by combination of antioxidant addition.