Disulfide Bond Formation Affects Stability of Whey Protein Isolate Emulsions

The viscosity and degree of flocculation of 20 wt% n-hexadecane oil-in-water emulsions stabilized by whey protein isolate (1 wt% WPI in 0.05M phosphate buffer, pH 7.0) increased as the emulsions aged. These effects were reduced when N-ethylmaleimide, a sulfhydryl blocking agent, was added to the emulsions immediately after homogenization, but were not completely eliminated. Gel electrophoresis (SDS-PAGE) showed an increase in the extent of intermolecular disulfide bond formation between proteins absorbed at the droplet interface with time. Floes were probably formed initially by noncovalent bonding or bridging flocculation, and then stabilized by disulfide bonds between proteins absorbed to different droplets.     

The Effect of Feed Solids Concentration and Inlet Temperature on the Flavor of Spray Dried Whey Protein Concentrate

Previous research has demonstrated that unit operations in whey protein manufacture promote off‐flavor production in whey protein. The objective of this study was to determine the effects of feed solids concentration in liquid retentate and spray drier inlet temperature on the flavor of dried whey protein concentrate (WPC). Cheddar cheese whey was manufactured, fat‐separated, pasteurized, bleached (250 ppm hydrogen peroxide), and ultrafiltered (UF) to obtain WPC80 retentate (25% solids, wt/wt). The liquid retentate was then diluted with deionized water to the following solids concentrations: 25%, 18%, and 10%. Each of the treatments was then spray dried at the following temperatures: 180 °C, 200 °C, and 220 °C. The experiment was replicated 3 times. Flavor of the WPC80 was evaluated by sensory and instrumental analyses. Particle size and surface free fat were also analyzed. Both main effects (solids concentration and inlet temperature) and interactions were investigated. WPC80 spray dried at 10% feed solids concentration had increased surface free fat, increased intensities of overall aroma, cabbage and cardboard flavors and increased concentrations of pentanal, hexanal, heptanal, decanal, (E)2‐decenal, DMTS, DMDS, and 2,4‐decadienal (P < 0.05) compared to WPC80 spray dried at 25% feed solids. Product spray dried at lower inlet temperature also had increased surface free fat and increased intensity of cardboard flavor and increased concentrations of pentanal, (Z)4‐heptenal, nonanal, decanal, 2,4‐nonadienal, 2,4‐decadienal, and 2‐ and 3‐methyl butanal (P < 0.05) compared to product spray dried at higher inlet temperature. Particle size was higher for powders from increased feed solids concentration and increased inlet temperature (P < 0.05). An increase in feed solids concentration in the liquid retentate and inlet temperature within the parameters evaluated decreased off‐flavor intensity in the resulting WPC80.     

酶解乳清浓缩蛋白WPC80制备乳清肽的研究

用乳清浓缩蛋白WPC80 为原料,在复合酶A 的作用下,研究乳清肽的制备工艺。复合酶A 的反应条件为[S]12g/100mL、温度50℃、[E]/[S]3%、pH9.0。选用截留分子质量10kD 的磺化聚砜膜,常温并在工作压差0.25MPa 下对水解液进行超滤处理后,选用树脂HZ00x 对水解液进行脱苦,得到的处理液无明显的苦涩味,只有轻微的蛋腥味,最后肽得率36.54%。产品中肽的分子质量分布以二、三和四肽为主,分别占峰面积的27.45%、34.88% 和26.65%。     

美拉德反应对乳清分离蛋白及其水解物抗氧化性的影响

以乳清分离蛋白及其蛋白水解物为原料分别与半乳糖发生美拉德反应,研究两者美拉德反应的褐变程度、接枝度及产物荧光光谱的变化,同时以乳清分离蛋白和蛋白水解物作对照,研究两者美拉德反应产物的抗氧化性。结果表明:乳清分离蛋白及其水解物美拉德反应的褐变程度、接枝度及产物的抗氧化性均在4 h达到最大。其中,乳清分离蛋白及其水解物与半乳糖美拉德反应的褐变程度和接枝度最大值分别为1.114、18.431%和1.413、28.273%;两者美拉德反应产物的还原力、2,2’-联氨-二(3-乙基-苯并噻唑-6-磺酸)二铵盐自由基清除能力和1,1-二苯基-2-三硝基苯肼自由基清除能力分别为0.605、46.29%、61.77%和0.923、69.81%、78.43%,均显著高于对照组(P0.05)。同时,内源荧光光谱发现,美拉德反应改变了乳清分离蛋白及其水解物的构象,使二者的结构更加松散。     

羟基自由基氧化对乳清蛋白疏水性的影响

本实验研究在羟基自由基氧化体系中,不同的H2O2 浓度(1～20mmol/L)、FeCl3 浓度(0.1～2mmol/L)以及不同的pH 值、温度、盐浓度等环境条件对乳清蛋白疏水性的影响,每种氧化条件的氧化时间分别为1、3、5h。结果表明,氧化和环境因素对乳清蛋白的疏水性影响很大,在H2O2 体系中当H2O2 浓度为5mmol/L 时疏水值达到最大,为9559.2,是对照组的4.88 倍,明显高于FeCl3 体系中的变化;疏水性在偏离等电点pH7.0 时较大,在适宜的温度和盐浓度下能够增加乳清蛋白的疏水性。说明氧化过程中,乳清蛋白的疏水性受H2O2 浓度的改变影响最大,并且也受其他环境条件的影响。     

酶法水解乳清分离蛋白制备纳米纤维

采用天冬氨酸内切酶(Asp endoproteinase,AspN)在37℃诱导乳清分离蛋白(whey protein isolate,WPI)制备蛋白质纳米纤维(protein nanofibrils,PNFs),考察WPI不同水解时间对产物PNFs形貌的影响。利用透射电子显微镜、原子力显微镜、小分子蛋白质十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、硫磺素T荧光、红外光谱以及激光散射法对PNFs及形成过程进行表征。结果显示,该方法制备的产物PNFs呈乳白色,AspN水解得到的5 kD左右的多肽为构筑单元,β-折叠为PNFs稳定存在的主要二级结构,PNFs平均粒径随水解时间延长而增大。酶法制备PNFs解决了酸法中的褐变问题,有望拓展其在食品领域的应用。     

Heat-induced Changes in Angel Food Cakes Containing Egg-white Protein or Whey Protein Isolate

ABSTRACT: Angel food cakes made from egg white or whey protein foams were compared. Cakes were evaluated based on final volume, dynamic volume change, and rheological transitions during baking. Cake expansion during baking was a function of protein concentration regardless of protein type. Cakes containing whey proteins had a lower ability to prevent collapse once starch gelatinization started during baking. Heat-treating whey proteins or adding xanthan gum increases cake volume, but not to the extent of egg-white proteins. Cakes containing egg-white proteins became more elastic at 60 to 85 °C than those containing whey proteins, indicating physical differences in the heat-set protein foam network associated with protein type.     

The Potential Source of B. licheniformis Contamination During Whey Protein Concentrate 80 Manufacture

The objective of this study was to determine the possible source of predominant Bacillus licheniformis contamination in a whey protein concentrate (WPC) 80 manufacturing plant. Traditionally, microbial contaminants of WPC were believed to grow on the membrane surfaces of the ultrafiltration plant as this represents the largest surface area in the plant. Changes from hot to cold ultrafiltration have reduced the growth potential for bacteria on the membrane surfaces. Our recent studies of WPCs have shown the predominant microflora B. licheniformis would not grow in the membrane plant because of the low temperature (10 °C) and must be growing elsewhere. Contamination of dairy products is mostly due to bacteria being released from biofilm in the processing plant rather from the farm itself. Three different reconstituted WPC media at 1%, 5%, and 20% were used for biofilm growth and our results showed that B. licheniformis formed the best biofilm at 1% (low solids). Further investigations were done using 3 different media; tryptic soy broth, 1% reconstituted WPC80, and 1% reconstituted WPC80 enriched with lactose and minerals to examine biofilm growth of B. licheniformis on stainless steel. Thirty‐three B. licheniformis isolates varied in their ability to form biofilm on stainless steel with stronger biofilm in the presence of minerals. The source of biofilms of thermo‐resistant bacteria such as B. licheniformis is believed to be before the ultrafiltration zone represented by the 1% WPC with lactose and minerals where the whey protein concentration is about 0.6%.     

乳清浓缩蛋白对松仁蛋白溶解度及其乳液稳定性的影响

为研究蛋白质-蛋白质相互作用对松仁蛋白(PKP)溶解度、结构和乳液性质的影响,以PKP和乳清浓缩蛋白(WPC)为研究对象,采用pH循环法制备PKP-WPC复合蛋白,利用SDS-PAGE、内源性荧光光谱、紫外-可见光谱、圆二色谱、荧光探针和ζ-电位等方法分析了复合蛋白结构特性和表面特性,再以PKP-WPC复合蛋白为原料,分别制备了油相体积分数为3%、10%和50%的乳液,并对制备的乳液性质进行了检测。结果表明:当WPC与PKP的质量比为1.0∶1.0,且体系pH值经历由7.0到12.0再回到7.0时的1次pH循环后,PKP的水溶性可从48.53%提高到92.43%。SDS-PAGE结果显示,PKP-WPC复合蛋白完整保留了PKP和WPC的亚基。内源性荧光光谱、紫外-可见光谱和圆二色谱结果表明,静电相互作用、疏水相互作用和氢键是驱动PKP和WPC相互作用的主要作用力,PKP与WPC相互作用使复合蛋白具有较高的结构韧性,抵抗酸诱导的构象折叠;WPC的加入改变了PKP的二级结构,α-螺旋、β-转角和无规卷曲结构的含量增加,而β-折叠结构相对含量降低。PKP-WPC复合蛋白具有较高的表面电荷(-34.74mV)来抵抗蛋白质的聚集。与由PKP制备的乳液相比,由PKP-WPC制备的乳液平均粒径和乳层析指数减小,ζ-电位绝对值增大,稳定性显著提高。乳液的性质因油相体积分数的不同而有较大的差异。油相体积分数为3%的复合乳液液滴小且分布均匀,稳定性好于油相体积分数为10%和50%的复合乳液。通过pH循环法,通过添加WPC,提高了PKP的溶解度,获得了稳定性较佳的PKP乳液,研究可为新型蛋白产品的研发提供理论基础,拓宽松仁蛋白在加工食品中的应用范围,推动PKP-WPC双蛋白乳液研究的发展。     

乳清浓缩蛋白对面条黏性的影响

在小麦粉中添加不同比例的乳清浓缩蛋白(Whey protein concentrate,WPC),并分析其对面条品质及黏性的影响,以达到增加营养价值及改善面条品质的作用。实验结果表明:WPC的添加可以明显改善面条的黏结现象,并降低其表面黏性,但较高的添加量则会降低小麦粉的面筋质量和湿面筋含量,并使干物质损失率和干物质吸水率升高;微观结构和TOM值的结果表明,WPC的添加使面条蛋白网络孔隙变大,减少了面条表面滞留或附着的淀粉,这可能是WPC改善面条黏结现象的原因。     

茶多酚和乳清蛋白对冷藏鱼糜保鲜效果的影响

为探究茶多酚和乳清蛋白的添加对冷藏鱼糜保鲜效果的影响,以草鱼鱼糜为研究对象,设计4 组进行实验,A组为对照组,B组添加0.05%茶多酚,C组添加5%乳清蛋白,D组添加0.05%茶多酚＋5%乳清蛋白。在4 ℃条件下分别冷藏0、1、3、5、7、9 d,通过测定总巯基含量、硫代巴比妥酸反应物值、过氧化值、挥发性盐基氮值、水分含量、凝胶强度的变化,研究茶多酚和乳清蛋白对冷藏鱼糜氧化及品质的影响。结果表明：与对照组相比,茶多酚的添加可以明显抑制蛋白氧化和脂肪氧化（P＜0.05）,抗氧化效果是D组＞B组＞C组;而乳清蛋白的添加可以明显减少鱼糜水分含量损失和凝胶劣化（P＜0.05）,其作用效果是D组＞C组＞B组。与冷藏最初相比,从3 d起,所有指标变化显著（P＜0.05）,且结果都显示同时添加茶多酚和乳清蛋白协同作用效果最显著;冷藏7 d时,D组的挥发性盐基氮值降低了4.3 mg/100 g,凝胶强度增加了260.07 g·mm,且保鲜效果显著（P＜0.05）。因此,茶多酚和乳清蛋白共同应用于鱼糜制品生产中,将更利于提高保鲜效果。     

低酯果胶和乳清分离蛋白复合凝胶性质的研究

本实验研究混合体系pH 值、NaCl 浓度和CaCl2 浓度对低酯果胶与乳清蛋白复合凝胶的硬度和持水能力的影响。响应面分析结果表明,pH 值、NaCl 浓度和CaCl2 浓度对凝胶性质有显著影响。低酯果胶和乳清分离蛋白复合凝胶的最佳条件为pH6、NaCl 浓度0.2mol/L、CaCl2 浓度10mmol/L,在此条件下制得的复合凝胶强度接近200g,持水能力接近75%。     

乳清分离蛋白水解物对冷藏鱼糜的保鲜作用

通过测定不同贮藏时间内(0 d、1 d、3 d、5 d、7 d、9 d)草鱼鱼糜的羰基含量、硫代巴比妥酸反应物(thiobarbituric acid-reactive substances,TBARS)、细菌总数、挥发性盐基氮(total volatile basic nitrogen,TVB-N)值、三甲胺(trimethylamine,TMA)值、K值等指标的变化,并分析各指标之间的相关性,研究冷藏(4℃)条件下乳清分离蛋白(WPI)水解物对草鱼鱼糜的抗氧化及保鲜效果。实验分为5组,第1组为对照组,第2~4组中分别加入2%、4%和6%的水解物,第5组中加入0.02%的BHA。结果表明:随着WPI水解物添加量的增加,鱼糜的蛋白和脂肪氧化抑制显著(p0.05),保鲜效果明显改善,其中6%WPI水解物处理组效果最明显,在贮藏9 d内,与对照组相比,羰基含量、TBARS值、TVB-N值和K值分别降低了1.54 nmo L/mg蛋白、0.21 mg/kg、5.84×10~(-2) mg/g和11.7%,此时鱼糜仍处于二级鲜度。各指标间存在显著正相关(p0.05)。因此,WPI水解物可以起到很好的抗氧化和保鲜作用,未来可以考虑把乳清蛋白水解物作为天然保鲜剂用于鱼糜制品生产中。     

High Hydrostatic Pressure Affects Flavor-binding Properties of Whey Protein Concentrate

ABSTRACT: The effects of high hydrostatic pressure (HHP) on flavor-binding properties of whey protein concentrate (WPC) were determined with benzaldehyde, heptanone, octanone, and nonanone. After HHP treatment (600 MPa, 50 °C, for 0-, 10-, or 30-min holding time), flavor-binding properties of WPC were studied by intrinsic fluorescence titration and static headspace analysis. The HHP treatments increased the number of binding sites and the apparent dissociation constants of WPC for benzaldehyde. HHP treatment of WPC for 0 min increased the number of binding sites of WPC for heptanone and octanone. As observed by headspace analysis, HHP treatments did not result in significant changes in the flavor retention for benzaldehyde in WPC solutions. Flavor retention of 100 ppm and 200 ppm heptanone and octanone in HHP-treated (10 min) WPC was significantly lower than for untreated WPC and HHP-treated WPC for 0 min or 30 min. For flavor retention of nonanone, significant decreases were only observed at 100 ppm when WPC solutions were HHP-treated for 10 min. While use of HHP treatment of WPC has potential in real food systems, these findings demonstrate the importance of careful selection of HHP treatment times and flavor concentrations for desired outcomes in food applications.     

乳铁蛋白-乳清分离蛋白乳状液微聚集体构建与酶交联对其流变学特性的影响

以乳清分离蛋白（whey protein isolate,WPI）与乳铁蛋白（lactoferrin,LF）乳状液形成微聚集体与转谷氨酰胺酶酶促交联微聚集体,以期提高体系流变特性。通过微射流分别制备WPI和LF乳状液,二者混合后,乳状液微滴之间发生异型聚集效应,通过转谷氨酰胺酶交联结合形成具有特定三维空间网络结构的微聚集体。研究结果表明：WPI与LF乳状液发生异型聚集,最大程度的聚集和最高物理稳定性体系发生在50% LF-50% WPI微滴形成的微聚集体。异型聚集效应改变了乳状液的流变特性,与单一WPI和LF乳状液相比,50% LF-50% WPI微聚集体流变学特性黏度值分别为单一乳状液的3.72?倍和2.2?倍,通过转谷氨酰胺酶交联,乳状液微聚集体的黏度值为原来的11.4?倍。因此,基于异型聚集效应结合酶促交联,可提高食品体系的流变特性,为开发食品脂质替代物提供了一定的理论支持。     

Edible Coatings on Frozen King Salmon: Effect of Whey Protein Isolate and Acetylated Monoglycerides on Moisture Loss and Lipid Oxidation

Whey protein isolate (WPI) and acetylated monoglyceride (AMG) coatings were evaluated for effectiveness against moisture loss and lipid oxidation of frozen King salmon. A model gel material was also used to screen coatings for effectiveness against moisture loss. Coatings of low-melting-point AMG used alone or after applying WPI solution or WPI powder were effective in reducing the rate of moisture loss by 42–65% during the first 3 wk of storage. Onset of lipid oxidation was delayed and peak peroxide values were reduced in samples coated with WPI solution/antioxidant overspray or those containing low-melting-point AMGs. No differences in effectiveness were found among the coating treatments.     

Whey Protein Isolate and Glyco-macropeptide Recovery from Whey Using Ion Exchange Chromatography

ABSTRACT: Cation exchange was used to recover whey protein isolate (WPI) from sweet whey, and the effluent was fed to an anion exchanger to recover glycomacropeptide (GMP). Nearly all of the major whey proteins (α-lactalbu-min, β-lactoglobulin, immunoglobulin G, and serum albumin) and about half of the total Kjeldahl nitrogen (TKN) were recovered by the cation exchanger. No GMP was recovered by the cation exchanger. The anion exchanger recovered nearly all of the GMP from the effluent of the cation exchanger, accounting for about half of the remaining TKN. This process is the first to simultaneously manufacture WPI and GMP from a single stream of whey, increasing the value obtained from whey.     

自由基氧化的乳清蛋白在冰淇淋中的应用

研究经羟基自由基产生体系氧化后的乳清蛋白在冰淇淋中的应用,测定蛋白氧化的主要指标羰基含量,并对影响冰淇淋品质的主要参数(包括膨胀率、抗融性、保形性)以及感官指标进行评定。结果表明：在冰淇淋中添加经H2O2体系氧化后的乳清蛋白,产品各项指标明显好于添加FeCl3体系的冰淇淋,且能够提高产品的性能,尤其是添加氧化1h的氧化蛋白其感官评分达到最大,但同对照比口感略有降低。这说明,适当的氧化可以改进产品的品质,提高产品的可接受性,但需改进口感。