A new method for determination of antithrombotic activity of egg white protein hydrolysate by microplate reader

A new method for the determination of antithrombotic activity of egg white protein hydrolysate （EWPH） was developed using a microplate reader. Reaction was carried out at 37℃and pH 7.2 with fibrinogen concentration 0.1%. Microplate reading was conducted at 405 nm. Inhibition rate of EWPH on thrombin activity showed linearity （R2 = 0.9971）, when the inhibition rate was in the range of 10-90%. The lower limit of detection （LLD, at 99.7% probability） and the biological limit of detection （BLD, at 99.7% probability） of the method were 10.643 and 40 mg/mL, respectively. The repeatability standard deviation （R.S.D.） was 1.08%. The standard deviation of the method was ±0.027 AT-U.

Regulation of Microstructure of Calcium Carbonate Crystals by Egg White Protein

Crystal growth of calcium carbonate in biological simulation was investigated via egg white protein with different volume fractions,during which calcium carbonate was synthesized by calcium chloride and sodium carbonate.The morphology,thermal properties and microstructure of the calcium carbonate micro-to-nanoscale crystals were characterized by scanning electron microscopy（SEM）,transmission electron microscopy（TEM）,Fourier transform infrared spectroscopy（FTIR）,thermogravimetric analysis（TG） and X-ray diffraction（XRD） analysis.The results show that the volume fraction of egg white protein has great influence on the shape,size and morphology of calcium carbonate crystals.The calcium carbonate crystals were the mixtures of calcite-vaterite-like crystals including spherical and rough surface,which are different from that formed in pure water.With the increase of egg white protein concentration,the diameter of calcium carbonate crystals changed,the amount of formed spherical calcium carbonate particles decreased and that of vaterite increased.These results indicate that the coordination and electrostatic interaction between egg white protein and Ca2＋ significantly affect the calcium carbonate crystalization.

Phase Transformation of Amorphous Calcium Carbonate to Single-Crystalline Aragonite with Macroscopic Layered Structure in the Presence of Egg White Protein and Zinc Ion

Highly oriented calcium carbonate lamellas are exquisite structure produced by biomineralization. Strategies mimicking nature have been developed to synthesize inorganic materials with excellent structures and optimal properties. In our strategy, egg white protein and zinc ion were employed in the solution to induce the crystallization of calcium carbonate, resulting in the macroscopic aragonite laminate with an average length of 1.5 mm, which was comprised of single-crystalline tablets. During the crystallization at initial stage, it was found that the particles displayed the characteristics of amorphous calcium carbonate, which was then transformed into the sophisticated structured aragonite through a multistage assembly process. The rebuilt nacre structure in vitro was achieved owing to the synergistic effects of egg white protein and zinc ion.

Effects of freeze-drying and spray drying processes on functional properties of phosphorylation of egg white protein

Egg white protein(EWP)was phosphorylated with Sodium Tripolyphosphate(STP)at pH 4.5.Freeze drying and spray drying were used for drying purpose and the effects of these drying methods on the functional properties were investigated.The functional properties of native and modified proteins were also determined.The results demonstrated that phosphorylation of EWP markedly improved its functional properties,and that it was more effective for the food industry.The freeze-dried STP-EWP powders were superior in terms of solubility,emulsion stability,water holding capacity,oil and water absorption capacity and heat gel strength than spray-dried STP-EWP powders.The results in viscosity showed no significant differences between freeze-dried and spray dried.The spray-dried powders were better in terms of foaming ability and foam stability than freeze-dried powders.However,the spray drying required the longest time to produce.Freeze drying was found to be the best method in terms of production of modified EWP powders with superior functional properties.

Aspergillus oryzae reduces IgE binding ability of allergenic egg white proteins

Egg white proteins are one of the major allergens. The objective of this study was to investigate the effect of Aspergillus oryzae cultivation on IgE binding ability of egg white proteins. Effect of A. oryzae on egg white proteins was determined using ninhydrin method,SDS-PAGE, ELISA, fluorescence FITC labeling, MALDITOF-MS and LC-MS/MS analysis. Adding mycelium of A. oryzae ATCC 1011 and 16868 substantially reduced the IgE binding ability of acidified egg white after 24 h incubation. The binding capacity of egg white proteins to IgE in plasma from four egg allergy patients was almost completely lost after incubation with mycelium of ATCC16868. Results from SDS-PAGE, free amino acid analysis,MALDI-TOF-MS and LC-MS/MS indicated that there was no substantial protein degradation during incubation.Therefore, the reduction of IgE binding ability of egg white proteins during A. oryzae treatment was probably due to a loss of ～1700 Da mass including a fragment of the ovomucoid N terminus.

Fast separation of hen egg white protein with a phosphorylcholine type zwitterionic ion chromatography stationary phase

In this work, a kind of preparation method of zwitterionic ion chromatography （ZIC） stationary phase modified with phosphorylcholine （PC） was obtained by hydrolyzing after bonding phosphorylcholine dichloride to diol-silica to better explore the characteristics of the PC groups as ZIC stationary phase ligand in simultaneous separation of acidic proteins and basic proteins. The results showed that tv^o kinds of acidic proteins and three kinds of basic proteins can be separated completely, meanwhile, hen egg white was separated and purified and three kinds of egg white components ovalbumin, G2 ovoglobulin and ovotransfemin proteins were separated completely by one single step on PC-ZIC column, the purity of all proteins reached above 95%. PC-ZIC stationary phase was successfully improved with better separation capacity and selectivity than previously reported in this paper.

Interaction mechanism of egg white-derived ACE inhibitory peptide TNGIIR with ACE and its effect on the expression of ACE and AT1 receptor

The egg white-derived hexapeptide TNGIIR inhibits angiotensin-converting enzyme(ACE)activity in vitro.In this work,molecular docking revealed that TNGIIR established hydrogen bonds with the S1(Ala 354),S2(Gln 281,His 513,Tyr 520 and Lys 511)and S1(Glu 162)pockets of ACE.In addition,the potential antihypertensive effect of the oral administration of TNGIIR in spontaneously hypertensive rats(SHR)was investigated,as was the effect of this peptide on the mRNA expression of ACE and angiotensin type 1(AT1)and type 2(AT2)receptors in renal tissue.The oral administration of TNGIIR(2,10 and 50 mg/kg)for up to four weeks did not reduce the blood pressure of SHR,in contrast to captopril(10 mg/kg,orally),but attenuated the mRNA expression of ACE and AT1 receptor(as did captopril).In contrast,both TNGIIR and captopril enhanced the expression of AT2 receptor mRNA.There was no change in the circulating concentration of angiotensin I,but a slight decrease(about 10%)was seen in the concentration of circulating angiotensin II with TNGIIR and captopril.

超高压协同TG酶改性蛋清蛋白热诱导凝胶机理

为提高蛋清蛋白凝胶性,本研究以蛋清蛋白(Egg white protein,EWP)为研究对象,通过分析质构、持水率、分子作用力、傅里叶红外色谱、圆二色谱、粒径、电位、巯基含量、表面疏水力及扫描电镜的变化,探究超高压处理(Ultra-high pressure,UHP)、谷氨酰胺转胺酶处理(Transglutaminase,TG)及超高压协同TG酶处理(Ultra-high pressure synergistic Transglutaminase,UTG)的蛋清蛋白热诱导凝胶机理及结构的变化。结果表明:UHP-EWP、TG-EWP、UTG-EWP的硬度、弹性和持水性均有所提高,疏水相互作用力是维持凝胶的主要作用力;UHP-EWP、TG-EWP、UTG-EWP的α-螺旋含量均不同程度下降、β-折叠含量均上升;UHP-EWP的平均粒径值下降、电位绝对值下降,TG-EWP、UTG-EWP的变化与之相反;UHP-EWP、TG-EWP、UTG-EWP的游离巯基含量上升、总巯基含量下降、表面疏水性升高;凝胶结构更加致密光滑,平整度提高。本研究为蛋清蛋白热诱导凝胶改性提供了理论基础及研究思路。

蛋清蛋白对低脂凝固型酸奶品质的影响

该研究以蛋清蛋白作为品质改良剂制备低脂凝固型酸奶,探究乳蛋白与蛋清蛋白比例、总蛋白质含量和蔗糖含量对凝固型酸奶pH、硬度、流变学特性、水分状态及感官品质的影响。蛋清蛋白添加比例的升高使得酸奶pH、硬度、储能模量、表观黏度和持水性增加。总蛋白质量分数2.5%时无法形成凝固酸奶,总蛋白质量分数为3.5%和4.5%的酸奶能良好凝固。添加蔗糖能够改善酸奶的感官品质,结果显示添加蔗糖质量分数为6%的酸奶具有最高的感官评分。相关性分析表明,酸奶质地与其储能模量、pH呈正相关,与持水能力呈负相关。这些结果表明,蛋清蛋白能够改善低脂凝固型酸奶的品质。

蛋清蛋白微粒基低脂沙拉酱流变学特征及品质优化

本实验以蛋清蛋白微粒(egg white protein microparticles,EWPM)为代脂基质,研究EWPM/黄原胶(xanthan gum,XG)共混质量分数、pH值、氯化钠含量、糖含量、均质条件等对超低脂沙拉酱产品流变学特性的影响规律。研究发现,XG对EWPM基超低脂沙拉酱具有显著的增稠效果,然而含量过高会导致低脂沙拉酱恢复性下降。基础调味组分对超低脂沙拉酱品质的影响存在较大差异,pH值和盐对体系黏度影响较大,糖添加量对黏度的影响相对较小。适度的胶体磨均质转速和均质时间可以减小产品的粒径,增加其内部均一性,显著提高产品的整体黏度。本实验以沙拉酱流变学行为调控为重点,为低脂沙拉酱的研究提供一个更加明确的工艺优化方向。

蛋清蛋白絮凝处理脱除玉米秸秆水解液中的木质素衍生物

为了提升玉米秸秆水解液(corn stalks hydrolysate,CSH)的可生化降解性,研究了蛋清蛋白(egg white protein,EWP)絮凝处理对CSH中木质素衍生物去除的影响,并与氯化铁、阳离子聚丙烯酰胺等常规絮凝剂的效果进行对比。结果表明,在初始CODCr浓度为40 g/L的CSH中使用EWP进行热絮凝处理(100℃下加热5 min),CSH中大分子木质素(以浊度计算)去除率达(76.87±3.00)%,显著优于氯化铁、阳离子聚丙烯酰胺的处理结果 (P <0.05)。同时,EWP热絮凝处理未造成糖、呋喃及弱酸等化合物明显损失。这是由于EWP通过疏水反应与木质素衍生物结合,进一步通过巯基-二硫键交联形成大分子化合物,促进了木质素衍生物从CSH中去除。

蛋清蛋白/黄油复合凝胶的特性研究

以蛋清蛋白(egg white protein,EWP)和黄油为原料,按照不同比例(1∶1、1∶2、1∶3、1∶4、1∶5,质量比)进行复配,经热诱导制备EWP/黄油复合凝胶,并分析其质构特性、持水性、乳化性、乳化稳定性、流变特性、扫描电镜及激光共聚焦的变化规律。结果表明,质构特性中的硬度及胶黏性随着黄油添加量的增加而增加,而弹性及内聚性则在EWP/黄油为1∶3(质量比)时达到最高(P<0.05);复合凝胶的持水性随着黄油添加量的增加而增加,在1∶4及1∶5(质量比)时差异不显著(P>0.05);当二者质量比为1∶3时,乳化性及乳化稳定性显著高于其他组(P<0.05);流变结果显示当二者质量比为1∶3时,复合凝胶具有较大的储能模量(G′)及较低的损耗因子tanδ,说明此比例下的黄油提高了复合凝胶的黏弹性;扫描电镜结果表明二者质量比为1∶3时促使复合凝胶形成了更致密紧凑的网络结构,而未添加黄油的蛋清蛋白凝胶则呈现孔洞较大且大小不均匀的结构特征;激光共聚焦结果显示EWP/黄油1∶3(质量比)时网络结构相对均匀,且无析水无孔隙。综上所述,EWP和黄油的质量比为1∶3时可以改善蛋清蛋白的乳化特性及凝胶特性,为后续替代食品中部分乳化盐,以降低食品中钠含量,制备相关低钠食品提供了数据支撑。

蛋清热凝胶透明机制研究进展

蛋白基凝胶由于其光学透明属性在食品和医药领域具有独特优势而倍受关注,透明凝胶有望成为类似于燕窝的高端蛋白基凝胶食品或其加工辅基(料),探明蛋清热凝胶光学性质的差异机制有望成为攻克蛋清凝胶高质化利用的关键。文章对蛋清凝胶透明度的形成机制及其影响因素进行综述。深入分析了蛋清热凝胶的形成机理,探讨了蛋白质的改性方法,并对蛋清透明热凝胶未来的研究发展方向进行了展望。

羧甲基纤维素的添加对蛋清蛋白功能特性的协同效应的影响

为了提高蛋清蛋白的功能性质,分别向蛋清蛋白溶液中加入0.2‰、0.4‰、0.6‰、0.8‰、1.0‰的羧甲基纤维素(CMC),研究蛋白-多糖混合体系的流变、质构、起泡性和差示热量扫描分析的变化。结果表明:羧甲基纤维素添加到蛋清蛋白溶液提高了起泡性,0.8‰CMC-蛋清蛋白泡沫稳定性最优。对于其回复性、咀嚼性、硬度、凝聚力和弹性也呈增强趋势,0.8‰CMC-蛋清蛋白的硬度和咀嚼性最优,其他性能均与1.0‰CMC-蛋清蛋白差别不大。热性能和流变性能表明CMC的加入提高了共混体系的热稳定性,0.8‰CMC-蛋清蛋白的溶胶转变温度达到最大值。

鸡蛋清中抑菌活性蛋白作用机制的研究进展

鸡蛋清中富含多种具有抑菌功能的生物活性成分。随着鸡蛋贮存时间延长,蛋清中的浓蛋白会发生稀化现象,从而导致抑菌效果降低,微生物繁殖加快,鸡蛋品质下降。本文对鸡蛋贮藏过程中的卵黏蛋白、溶菌酶及其复合物的相互作用形式、抑菌机制等进行探讨,以期为优化鸡蛋加工过程中的消毒杀菌工序、改良鸡蛋贮藏方法、延长鸡蛋保质期提供参考。

不同类型蛋白对白令海狭鳕鱼糜凝胶特性的影响

研究金线鱼(Nemipterus virgatus)、蛋清蛋白、大豆蛋白粉、南美白对虾(Litopenaeus vannamei)和鲢(Hypophthalmichthys molitrix)5种不同类型蛋白的肉类添加对白令海狭鳕(Theragra chalcogramma)鱼糜凝胶性能的影响。在鱼糜中分别添加1%、3%、5%、7%、9%的金线鱼、蛋清蛋白、大豆蛋白粉、南美白对虾和鲢鱼肉以制备鱼糜凝胶制品并测定其白度、持水性、凝胶强度等品质指标。研究结果表明,适量添加金线鱼、乳清蛋白和大豆蛋白粉能有效地改善鱼糜的凝胶强度和持水性,但大豆蛋白粉的添加会降低鱼糜白度。乳清蛋白的添加对鱼糜的凝胶强度和持水性的提高效果最好,添加量为3%时达到最大值,分别为12301.34 g·mm和73.21%,蒸煮损失率仅为12.6%,说明该比例的添加可以最大程度地提高白令海狭鳕鱼糜凝胶的品质。在实际鱼糜制品的生产中,应根据产品的特征选择适合的蛋白种类和合理的添加量,来提高鱼糜制品的品质。该研究为白令海狭鳕鱼糜制品的开发提供理论依据与参考。

Imaging protein crystal growth behaviour in batch cooling crystallisation

The temporal and spatial growth behaviour of protein crystals, subject to different cooling strategies in protein crystallisation was investigated. Although the impact of temperature and cooling rate on crystal growth of small molecules was well documented, much less has been reported on their impact on the crystallisation of proteins. In this paper, an experimental set-up is configured to carry out such a study which involves an automatic temperature controlled hot-stage crystalliser fitted with a real-time imaging system. Linbro parallel crystallisation experiments(24-well plate) were also conducted to find the suitable initial conditions to be used in the hot-stage crystallisation experiments, including the initial concentration of HEW lysozyme solutions, precipitate concentration and pH value. It was observed that fast cooling rates at the early stage led to precipitates while slow cooling rates produced crystal nuclei, and very slow cooling rates, much smaller than for small molecules are critical to the growth of the nuclei and the crystals to a desired shape. The interesting results provide valuable insight as well as experimental proof of the feasibility and effectiveness of cooling as a means for achieving controlled protein crystallisation, compared with the evaporation approach which was widely used to grow single large crystals for X-ray diffraction study. Since cooling rate control can be easily achieved and has good repeatability, it suggests that large-scale production of protein crystals can be effectively achieved by manipulating cooling rates.

蛋清蛋白凝胶改性及其在肉制品加工中的应用

蛋清蛋白具有凝胶性、起泡性、乳化性等功能特性,生产加工中通常采用改性处理的方法提高蛋清蛋白的凝胶特性及凝胶稳定性。文章综述了蛋清蛋白的改性方法以及改性蛋清蛋白在肉制品加工中应用的研究现状,并对蛋清蛋白改性方法的创新及应用进行了展望。

湿法糖基化改性对蛋清蛋白凝胶特性及微观结构的影响

以蛋清蛋白(Egg White Protein,EWP)为主要原料,研究低聚半乳糖(Galactooligosaccharides,GOS)改性对蛋清蛋白凝胶性能的影响,并探讨其凝胶机理。首先研究了GOS添加量及热诱导条件对复合凝胶硬度和弹性的影响,然后通过低场核磁、流变仪、傅里叶红外光谱、扫描电镜等手段对复合凝胶的色差、保水性、流变特性、分子间作用力及微观结构进行分析,探究糖基化改性提高EWP凝胶特性机理。结果表明,GOS添加量及热诱导条件能够显著(P<0.05)影响复合凝胶的质构特性,与EWP凝胶相比,在最佳条件下制得的复合凝胶硬度和弹性分别增加68.30%和15.05%,白度下降23.37%,保水性增加27.54%。EWP经糖基化改性后,疏水相互作用及二硫键贡献增强,GOS碳氧双键与EWP伯氨基发生共价交联,使凝胶的微观结构更加致密、保水性更强。本研究为蛋清蛋白湿法糖基化改性的制备及应用提供理论基础。

低聚糖改性对蛋清蛋白凝胶性影响研究进展

蛋清蛋白是一种营养价值丰富,具有多种功能特性的优质蛋白。低聚糖可通过糖基化反应与蛋清蛋白共价结合,其偶联物的功能特性较天然蛋清蛋白有所改善。文章综述了低聚糖-蛋清蛋白偶联物的凝胶机理,探讨了低聚糖改性蛋清蛋白凝胶的影响因素,梳理了低聚糖对蛋清蛋白功能特性产生的影响,并对低聚糖改性蛋清蛋白的加工应用进行了展望。