Application of whey protein isolate in bone regeneration: Effects on growth and osteogenic differentiation of bone-forming cells

Recently, milk-derived proteins have attracted attention for applications in the biomedical field such as tissue regeneration. Whey protein isolate (WPI), especially its main component β-lactoglobulin, can modulate immunity and acts as an antioxidant, antitumor, antiviral, and antibacterial agent. There are very few reports of the application of WPI in tissue engineering, especially in bone tissue engineering. In this study, we tested the influence of different concentrations of WPI on behavior of human osteoblast-like Saos-2 cells, human adipose tissue-derived stem cells (ASC), and human neonatal dermal fibroblasts (FIB). The positive effect on growth was apparent for Saos-2 cells and FIB but not for ASC. However, the expression of markers characteristic for early osteogenic cell differentiation [type-I collagen (COL1) and alkaline phosphatase (ALP)] as well as ALP activity, increased dose-dependently in ASC. Importantly, Saos-2 cells were able to deposit calcium in the presence of WPI, even in a proliferation medium without other supplements that support osteogenic cell differentiation. The results indicate that, depending on the cell type, WPI can act as an enhancer of cell proliferation and osteogenic differentiation. Therefore, enrichment of biomaterials for bone regeneration with WPI seems a promising approach, especially due to the low cost of WPI.     

以乳清蛋白为基质的脂肪替代品对冰淇淋品质的影响

以乳清蛋白WPC-80为基质制备脂肪替代品,替代中脂冰淇淋中25%的脂肪。随着脂肪替代率的增加,冰淇淋浆料黏度和膨胀率增大,抗融化率和硬度下降,制得的低脂冰淇淋的各项感官指标均与中脂冰淇淋相当。当替代全部脂肪时,所制得的无脂冰淇淋也有较好的感官接受性。     

Astringency of bovine milk whey protein

Whey protein solutions at pH 3.5 elicited an astringent taste sensation. The astringency of whey protein isolate (WPI), the process whey protein (PWP) that was prepared by heating WPI at pH 7.0, and the process whey protein prepared at pH 3.5 (aPWP) were adjusted to pH 3.5 and evaluated by 2 sensory analyses (the threshold method and the scalar scoring method) and an instrumental analysis (taste sensor method). The taste-stimulating effects of bovine and porcine gelatin were also evaluated. The threshold value of astringency of WPI, PWP, and aPWP was 1.5, 1.0, and 0.7 mg/mL, respectively, whereas the gelatins did not give definite astringency. It was confirmed by the scalar scoring method that the astringency of these proteins increased with the increase in protein concentration, and these proteins elicited strong astringency at 10 mg/mL under acidic conditions. On the other hand, the astringency was not elicited at pH 3.5 by 2 types of gelatin. A taste sensor gave specific values for whey proteins at pH 3.5, which corresponded well to those obtained by the sensory analysis. Elicitation of astringency induced by whey protein under acidic conditions would be caused by aggregation and precipitation of protein molecules in the mouth.     

Efficiency of removal of whey protein from sweet whey using polymeric microfiltration membranes

Our objective was to measure whey protein removal percentage from separated sweet whey using spiral-wound (SW) polymeric microfiltration (MF) membranes using a 3-stage, 3× process at 50°C and to compare the performance of polymeric membranes with ceramic membranes. Pasteurized, separated Cheddar cheese whey (1,080 kg) was microfiltered using a polymeric 0.3-μm polyvinylidene (PVDF) fluoride SW membrane and a 3×, 3-stage MF process. Cheese making and whey processing were replicated 3 times. There was no detectable level of lactoferrin and no intact α- or β-casein detected in the MF permeate from the 0.3-μm SW PVDF membranes used in this study. We found BSA and IgG in both the retentate and permeate. The β-lactoglobulin (β-LG) and α-lactalbumin (α-LA) partitioned between retentate and permeate, but β-LG passage through the membrane was retarded more than α-LA because the ratio of β-LG to α-LA was higher in the MF retentate than either in the sweet whey feed or the MF permeate. About 69% of the crude protein present in the pasteurized separated sweet whey was removed using a 3×, 3-stage, 0.3-μm SW PVDF MF process at 50°C compared with 0.1-μm ceramic graded permeability MF that removed about 85% of crude protein from sweet whey. The polymeric SW membranes used in this study achieve approximately 20% lower yield of whey protein isolate (WPI) and a 50% higher yield of whey protein phospholipid concentrate (WPPC) under the same MF processing conditions as ceramic MF membranes used in the comparison study. Total gross revenue from the sale of WPI plus WPPC produced with polymeric versus ceramic membranes is influenced by both the absolute market price for each product and the ratio of market price of these 2 products. The combination of the market price of WPPC versus WPI and the influence of difference in yield of WPPC and WPI produced with polymeric versus ceramic membranes yielded a price ratio of WPPC versus WPI of 0.556 as the cross over point that determined which membrane type achieves higher total gross revenue return from production of these 2 products from separated sweet whey. A complete economic engineering study comparison of the WPI and WPPC manufacturing costs for polymeric versus ceramic MF membranes is needed to determine the effect of membrane material selection on long-term processing costs, which will affect net revenue and profit when the same quantity of sweet whey is processed under various market price conditions.     

Impact of whey protein isolates and concentrates on the formation of protein nanoparticles‐stabilised Pickering emulsions

Whey protein nanoparticles (NPs) were prepared by heat‐induced method. The influences of whey protein isolates (WPIs) and concentrates (WPCs) on the formation of NPs were first investigated. Then Pickering emulsions were produced by protein NPs and their properties were evaluated. After heat treatment, WPC NPs showed larger particle size, higher stability against NaCl, lower negative charge and contact angle between air and water. Dispersions of WPC NPs appeared as higher turbidity and viscosity than those of WPI NPs. The interfacial tension of WPC NPs (~7.9 mN/m at 3 wt% NPs) was greatly lower than that of WPI NPs (~12.1 mN/m at 3 wt% NPs). WPC NPs‐stabilised emulsions had smaller particle size and were more homogeneous than WPI NPs‐stabilised emulsions. WPC NPs‐stabilised emulsions had higher stability against NaCl, pH and coalescence during storage.     

Production of a high gel strength whey protein concentrate from cheese whey

In order to develop a process for the production of a whey protein concentrate (WPC) with high gel strength and water-holding capacity from cheese whey, we analyzed 10 commercially available WPC with different functional properties. Protein composition and modification were analyzed using electrophoresis, HPLC, and mass spectrometry. The analyses of the WPC revealed that the factors closely associated with gel strength and water-holding capacity were solubility and composition of the protein and the ionic environment. To maintain whey protein solubility, it is necessary to minimize heat exposure of the whey during pretreatment and processing. The presence of the caseinomacropeptide (CMP) in the WPC was found to be detrimental to gel strength and water-holding capacity. All of the commercial WPC that produced high-strength gels exhibited ionic compositions that were consistent with acidic processing to remove divalent cations with subsequent neutralization with sodium hydroxide. We have shown that ultrafiltration/diafiltration of cheese whey, adjusted to pH 2.5, through a membrane with a nominal molecular weight cut-off of 30,000 at 15 degrees C substantially reduced the level of CMP, lactose, and minerals in the whey with retention of the whey proteins. The resulting WPC formed from this process was suitable for the inclusion of sodium polyphosphate to produce superior functional properties in terms of gelation and water-holding capacity.     

The effect of bleaching agent on the flavor of liquid whey and whey protein concentrate

The increasing use and demand for whey protein as an ingredient requires a bland-tasting, neutral-colored final product. The bleaching of colored Cheddar whey is necessary to achieve this goal. Currently, hydrogen peroxide (HP) and benzoyl peroxide (BPO) are utilized for bleaching liquid whey before spray drying. There is no current information on the effect of the bleaching process on the flavor of spray-dried whey protein concentrate (WPC). The objective of this study was to characterize the effect of bleaching on the flavor of liquid and spray-dried Cheddar whey. Cheddar cheeses colored with water-soluble annatto were manufactured in duplicate. Four bleaching treatments (HP, 250 and 500 mg/kg and BPO, 10 and 20 mg/kg) were applied to liquid whey for 1.5 h at 60°C followed by cooling to 5°C. A control whey with no bleach was also evaluated. Flavor of the liquid wheys was evaluated by sensory and instrumental volatile analysis. One HP treatment and one BPO treatment were subsequently selected and incorporated into liquid whey along with an unbleached control that was processed into spray-dried WPC. These trials were conducted in triplicate. The WPC were evaluated by sensory and instrumental analyses as well as color and proximate analyses. The HP-bleached liquid whey and WPC contained higher concentrations of oxidation reaction products, including the compounds heptanal, hexanal, octanal, and nonanal, compared with unbleached or BPO-bleached liquid whey or WPC. The HP products were higher in overall oxidation products compared with BPO samples. The HP liquid whey and WPC were higher in fatty and cardboard flavors compared with the control or BPO samples. Hunter CIE Lab color values (L*, a*, b*) of WPC powders were distinct on all 3 color scale parameters, with HP-bleached WPC having the highest L* values. Hydrogen peroxide resulted in a whiter WPC and higher off-flavor intensities; however, there was no difference in norbixin recovery between HP and BPO. These results indicate that the bleaching of liquid whey may affect the flavor of WPC and that the type of bleaching agent used may affect WPC flavor.     

Efficient encapsulation of fish oil: Capitalizing on the unique inherent characteristics of whey cream and hydrolyzed whey protein

The effects of protein concentration and of blending a phospholipid-rich whey coproduct, Procream (Salibra 700 Procream, Glanbia Nutritionals), with intact or hydrolyzed whey protein concentrate, on fish oil microencapsulation efficiency and oxidative stability were assessed. Trypsin and protease M, from Aspergillus oryzae, were used to produce 2 unique hydrolysates. All microcapsules had excellent encapsulation efficiencies (>92%) and good physical properties, regardless of protein content and Procream inclusion. Intact α-lactalbumin and β-lactoglobulin and their peptides were involved in stabilizing oil droplets. Disulfide interchange resulted in formation of protein aggregates, which were more pronounced in samples containing Procream. Although all microcapsules had relatively good oxidative stability, most had better stability at 2 versus 0.5% protein. Protease M hydrolysate + Procream microcapsules had the highest stability, regardless of protein content. Results demonstrated that Procream, at a reduced protein inclusion level, can partially replace more expensive whey protein ingredients in microencapsulation, when blended with a select hydrolysate.     

乳清多肽的分离纯化及体外抗氧化活性研究

采用乳酸菌发酵乳清制备乳清多肽,利用超滤和凝胶Sephadex G-25分离纯化乳清多肽,并研究了超滤后乳清多肽对羟自由基、氧自由基和DPPH自由基的清除作用。结果表明,超滤后乳清多肽主要包含两个级分,分子量分别为2 031 u和328 u,对羟自由基、氧自由基和DPPH自由基具有清除作用。     

乳清蛋白的功能特性及应用

介绍了乳清蛋白的组成成分和功能特性；论述了乳清蛋白的生物利用价值及在运动营养、医疗保健和食品加工等方面的广阔应用前景。     

Effect of whey concentration on protein recovery in fresh ovine ricotta cheese

Ricotta cheese, particularly the ovine type, is a typical Italian dairy product obtained by heat-coagulation of the proteins in whey. The aim of this work was to investigate the influence of whey protein concentration, obtained by ultrafiltration, on yield of fresh ovine ricotta cheese. Ricotta cheeses were obtained by thermocoagulation of mixtures with protein content of 1.56, 3.10, 4.16, and 7.09 g/100 g from the mixing of skim whey and ultrafiltered skim whey. A fat-to-protein ratio of 1.1 (wt/wt) was obtained for all mixtures by adding fresh cream. The initial mixtures, as well as the final ricotta cheeses, were analyzed for their composition and by SDS-PAGE. Protein bands were quantified by QuantityOne software (Bio-Rad, Hercules, CA) and identified by liquid chromatography-tandem mass spectrometry. Significant differences in the composition of the ricotta cheese were observed depending on protein concentration. Particularly, ricotta cheese resulting from the mixture containing 7.09 g/100 g of protein presented higher moisture (72.88 ± 1.50 g/100 g) and protein (10.18 ± 0.45 g/100 g) contents than that prepared from the mixture with 1.56 g/100 g of protein (69.52 ± 1.75 and 6.70 ± 0.85 g/100 g, respectively), and fat content was lower in this sample (12.20 ± 1.60 g/100 g) compared with the other treatments, with mean values between 15.72 and 20.50 g/100 g. Each protein fraction presented a different behavior during thermocoagulation. In particular, the recovery of β-lactoglobulin and α-lactalbumin in the cheese increased as their content increased in the mixtures. It was concluded that concentrating ovine rennet whey improved the extent of heat-induced protein aggregation during the thermal coagulation process. This resulted in a better recovery of each protein fraction in the product, and in a consequent increase of ricotta cheese yield.     

Effect of double homogenization and whey protein concentrate on the texture of ice cream

Ice cream samples were made with a mix composition of 11% milk fat, 11% milk solids-not-fat, 13% sucrose, 3% corn syrup solids (36 dextrose equivalent), 0.28% stabilizer blend, or 0.10% emulsifier and vanilla extract. Mixes were high temperature short time pasteurized at 80 degrees C for 25 s, homogenized at 141 kg/cm2 pressure on the first stage and 35 kg/cm2 pressure on the second, and cooled to 3 degrees C. The study included six treatments from four batches of mix. Mix from batch one contained 0.10% emulsifier. Half of this batch (treatment 1), was subsequently frozen and the other half (upon exiting the pasteurizer) was reheated to 60 degrees C, rehomogenized at 141 kg/cm2 pressure on the first stage and 35 kg/cm2 pressure on the second (treatment 2), and cooled to 3 degrees C. Mix from batch two contained 0.28% stabilizer blend. Half of this batch was used as the control (treatment 3), the other half upon exiting the pasteurizer was reheated to 60 degrees C, rehomogenized at 141 kg/cm2 pressure on the first stage and 35 kg/cm2 pressure on the second (treatment 4), and cooled to 3 degrees C. Batch three, containing 0.10% emulsifier and 1% whey protein concentrate substituted for 1% nonfat dry milk, upon exiting the pasteurizer was reheated to 60 degrees C, rehomogenized at 141 kg/cm2 pressure on the first stage and 35 kg/cm2 pressure on the second (treatment 5), and cooled to 3 degrees C. Batch four, containing 0.28% stabilizer blend and 1% whey protein concentrate substituted for 1% nonfat dry milk, upon exiting the pasteurizer was reheated to 60 degrees C, rehomogenized at 141 kg/ cm2 pressure on the first stage and 35 kg/cm2 pressure on the second (treatment 6), and cooled to 3 degrees C. Consistency was measured by flow time through a pipette. Flow time of treatment 3 was greater than all treatments, and the flow times of treatments 4 and 6 were greater than treatments 1, 2, and 5. Flow time was increased in ice cream mix by the addition of stabilizer. Double homogenization lowered ice cream mix flow time in the presence of stabilizer, but no difference in flow time was observed without stabilizer addition. Treatment 4 had a lower mean ice crystal size at 10 d postmanufacture compared with treatment 3; however, overall texture acceptability between treatments 3 and 4 was similar. Mean ice crystal size of treatment 6 was less at 18 wk postmanufacture compared with treatment 3; however, overall texture acceptability for treatments 3, 4, and 6 was similar. Mean ice crystal sizes of treatments 1, 2, and 5 were greater at 10 d and 18 wk compared with treatment 3. Sensory evaluation indicated that treatments 3, 4, and 6 had higher mean scores for icy, coldness intensity, and creaminess than treatments 1, 2, and 5 at 10 d and 18 wk postmanufacture.     

乳清多肽的制备及其发酵饮料的开发

研究了乳清多肽的制备、性质及其发酵饮料的开发,结果表明,碱性蛋白酶比中性蛋白酶水解乳清蛋白的能力强,且更经济,水解最佳条件为加酶量为7000(U/g蛋白)、底物添加量为5%、水解温度为60℃、水解初始pH值为8.5,最大乳清蛋白水解度可达到22.45%。最优酒精发酵条件为接种量5%、初始pH7.5、温度22℃、时间45h。乳清多肽发酵饮料的配方为酸量0.1%,蔗糖量为8%,-β环状糊精量为0.5%。     

乳清蛋白的功能成分及其主要应用

乳清蛋白是干酪生产的副产品,它具有神奇的功能成分,广泛应用于食品行业、医药、饲料、化妆品及其它行业。乳清蛋白的人气正在上升,它已经成为风靡全球的“健康代表”。     

大豆乳清蛋白乳化性的研究

对大豆乳清蛋白的乳化性进行了研究,并与大豆分离蛋白进行了比较。研究结果表明,大豆乳清蛋白的乳化稳定性优于大豆分离蛋白,乳化能力与大豆分离蛋白基本相当。正交实验确定大豆乳清蛋白乳化稳定性效果最优条件是:浓度20%、温度40℃、pH值9、时间25min。大豆乳清蛋白的乳化性的最佳条件是:浓度25%、温度40℃、pH值9、时间25min。离子对大豆乳清蛋白乳化性和乳化稳定性的影响为钠离子>钾离子,且钠离子的作用明显,使用大豆乳清蛋白乳化稳定性功能时选用离子条件是钠离子0.01mol/L,使用大豆乳清蛋白乳化性功能时选用离子条件是钠离子0.05mol/L。     

Factors regulating astringency of whey protein beverages

A rapidly growing area of whey protein use is in beverages. There are 2 types of whey protein-containing beverages: those at neutral pH and those at low pH. Astringency is very pronounced at low pH. Astringency is thought to be caused by compounds in foods that bind with and precipitate salivary proteins; however, the mechanism of astringency of whey proteins is not understood. The effect of viscosity and pH on the astringency of a model beverage containing whey protein isolate was investigated. Trained sensory panelists (n = 8) evaluated the viscosity and pH effects on astringency and basic tastes of whey protein beverages containing 6% wt/vol protein. Unlike what has been shown for alum and polyphenols, increasing viscosity (1.6 to 7.7 mPa·s) did not decrease the perception of astringency. In contrast, the pH of the whey protein solution had a major effect on astringency. A pH 6.8 whey protein beverage had a maximum astringency intensity of 1.2 (15-point scale), whereas that of a pH 3.4 beverage was 8.8 (15-point scale). Astringency decreased between pH 3.4 and 2.6, coinciding with an increase in sourness. Decreases in astringency corresponded to decreases in protein aggregation as observed by turbidity. We propose that astringency is related to interactions between positively charged whey proteins and negatively charged saliva proteins. As the pH decreased between 3.4 and 2.6, the negative charge on the saliva proteins decreased, causing the interactions with whey proteins to decrease.     

乳清蛋白的酶法改性研究

采用碱性蛋白酶、风味蛋白酶、木瓜蛋白酶、中性蛋白酶、复合蛋白酶、胰蛋白酶、胃蛋白酶在各自的最适条件下对乳清蛋白粉进行水解,通过HPLC方法测定酶解产物中α-乳白蛋白（α—La）和β-乳球蛋白（β—Lg）的含量。结果表明,碱性蛋白酶对其水解最快,其次为木瓜蛋白酶,而在胃蛋白酶和胰蛋白酶作用下则不易酶解,而且发现α-La比β—Lg更容易水解。     

Effect of heating temperature,pH, concentration and starch/whey protein ratio on the viscoelastic properties of corn starch/whey protein mixed gels

The viscoelastic properties of corn starch (CS) gels were more dependent on heating temperature, while the properties of whey protein isolate (WPI) gels were more dependent on pH. Thus heating temperature (75, 85, 95 °C) and pH (5, 7, 9) were varied to obtain a series of mixed gels with interesting viscoelastic properties. WPI gels showed extensive stress relaxation (SR) indicative of a highly transient network structure, while CS gels relaxed very little in 2000 s. Based on SR results, it appeared that CS/WPI mixed gels with 25 and 50% CS formed compatible network structures at 15% total solids only at pH 9. This supposition was supported by SEM microstructures obtained for dehydrated gels and a synergistic increase in the large‐strain fracture stress for these gels. Some synergy was also found for mixed gels at 30% total solids at pH 9, while at pH 7 the mixed gels seemed to contain separate additive WPI and CS networks unlike the case for pH 7 at 15% total solids. In both cases (15 and 30% total solids) the degree of elasticity of the mixed gels decreased as the WPI content increased. Mixed gels (CS:WPI = 0.5) at pH 9 showed increased fracture stress and fracture strain relative to the same gels at pH 7. This suggests that a unique chemical compatibility exists at pH 9 and results in gels that combine the elasticity of CS and the internal stress dissipation of WPI. © 2001 Society of Chemical Industry