大豆分离蛋白O/W型乳液膜的制备及其性质研究

摘要：为提高大豆分离蛋白膜的性质,制备以大豆分离蛋白（SPI）为原料的O/W乳液膜,采取单因素实验法比较O/W乳液膜与SPI膜的差异性,探究不同SPI质量浓度（20、30、40、50和60 mg/mL）对O/W乳液膜性能及结构的影响。结果表明：O/W乳液膜的遮光性、机械性能、耐水性及热稳定性要优于SPI膜。随着SPI质量浓度的增加,O/W乳液膜的拉伸强度随之增加,断裂伸长率降低。蛋白质量浓度为60 mg/mL时,O/W乳液膜的抗拉强度达到最大值为7.19 MPa,比蛋白膜的抗拉强度高出33%。利用扫描电子显微镜（SEM）、傅里叶红外光谱（FTIR）和差式扫描量热仪（DSC）对O/W乳液膜蛋白分子间的相互作用进行分析,乳液膜的Tg为100 ℃,且红外光谱中酰胺Ⅲ带和1630 cm-1峰值发生了变化,表明共混组分之间的作用力增强,蛋白质形成了致密且稳定的网络结构。     

Oil-in-Water Emulsions Stabilized by Sodium Caseinate or Whey Protein Isolate as influenced by Glycerol Monostearate

Competitive adsorption between glycerol monostearate (GMS) and whey protein isolate (WPI) or sodium caseinate was studied in oil-in-water emulsions (20 wt % soya oil, deionized water, pH 7). Addition of GMS resulted in partial displacement of WPI or sodium caseinate from the emulsion interface. SDS-PAGE showed that GMS altered the adsorbed layer composition in sodium caseinate stabilized emulsions containing < 1.0 wt % protein. Predominance of β-casein at the interface in the absence of surfactant was reduced in the presence of GMS. The distribution of α-lactalbumin and β-lactoglobulin between the aqueous bulk phase and the fat surface in emulsions stabilized with WPI was independent of the concentration of added protein or surfactant.     

Flocculation of Whey Protein Stabilized Emulsions as Influenced by Dextran Sulfate and Electrolyte

The creaming stability and viscosity of oil-in-water emulsions stabilized by whey protein isolate were monitored as functions of dextran sulfate (DS) and electrolyte (NaCl) concentration. At a specific DS concentration (the critical flocculation concentration, CFC), the droplets became flocculated, which promoted creaming. Addition of electrolyte caused an increase in CFC. At NaCl concentrations <0.5 wt%, addition of electrolyte decreased emulsion viscosity, but at concentrations >0.5 wt% it caused an increase in viscosity due to increased flocculation. The results were due to the influence of electrostatic screening on the effective volume of DS molecules and colloidal interactions between droplets.     

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.     

米糠蛋白O/W及W/O/W乳液制备及界面稳定性

为对比不同米糠蛋白质量浓度下O/W及W/O/W乳液的稳定性,以米糠蛋白作为基料,采用双乳化法制备O/W及W/O/W乳液,考察不同米糠蛋白质量浓度下乳液的微观形态和稳定性并探究其界面稳定机理。结果表明：W/O/W乳液的贮存稳定性显著优于O/W乳液;与相同蛋白含量的O/W乳液相比,W/O/W乳液的黏度显著提高;当米糠蛋白质量浓度为0.4 g/100 mL时,W/O/W乳液的稳定性较O/W乳液提高了1 倍以上;乳液内部包裹更多的W/O液滴,W/O/W乳液的粒径较大;而此时静电斥力也较大,起到稳定乳液的目的。同时,米糠蛋白质量浓度不小于0.4 g/100 mL时,O/W及W/O/W乳液中蛋白质的吸附率较高,达到78%以上。本研究为天然米糠蛋白质在食品级乳液中的开发提供参考,为粮食副产物的综合利用提供了新思路。     

Droplet Size and Coalescence Stability of Whey Protein Stabilized Milkfat Peanut Oil Emulsions

Droplet diameter and the polydispersity of droplet size tended to decrease with increased proportion of peanut oil. Macromolecular additives affected droplet size. Xanthan gum or sodium carboxymethyl-cellulose (Na-CMC) produced smaller droplets than the controls, but the average diameter was independent of the composition of the dispersed phase. At 50°C the coalescence stability of these emulsions qualitatively correlated with the initial droplet diameter. Xanthan gum and Na-CMC, despite increasing continuous phase viscosity, gave lower stability than controls, primarily the result of flocculation of droplets due to depletion of the highly hydrophilic macromolecule from the intervening region between approaching droplets. At 25°C, stability increased as solid butter oil content of the dispersed phase increased.     

大豆球蛋白-花青素Pickering乳液性质

制备大豆分离蛋白(soybean protein isolate,SPI)与花青素(anthocyanin,ACN)共价复合Pickering乳液。研究不同ACN体积分数下,共价复合颗粒的表面疏水性,Pickering乳液的乳化性与乳化稳定性、流变性质和微观结构。结果显示,当ACN体积分数由0%增加到0.15%时,共价复合颗粒的表面疏水性由18 174降低到8 945;Pickering乳液的乳化性增加了127 m2/g,乳化稳定性增加了近1倍;同时乳液脂滴状态得到了明显的改善。实验结果证明,乳液呈现类固体特性,表现出典型非牛顿假塑性行为。本研究还发现,随着ACN的添加,SPI-ACN共价复合Pickering乳液呈现出桥接乳液形态,这将为食品行业中开发新型Pickering乳液提供理论参考。     

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。     

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。     

Dispersion Stability of O/W Emulsions with Different Oil Contents Under Various Freezing and Thawing Conditions

Freezing and thawing of oil‐in‐water (O/W) emulsion‐type foods bring about oil–water separation and deterioration; hence, the effects of freezing and thawing conditions on the destabilization of O/W emulsions were examined. The freezing rate and thawing temperature hardly affected the stability of the O/W emulsion. O/W emulsions having different oil fractions were stored at temperatures ranging from –30 to –20 °C and then thawed. The stability after thawing depended on the storage temperature, irrespective of the oil fraction of the emulsion. A good correlation was found between the time at which the stability began to decrease and the time taken for the oil to crystalize. These results indicated that the dominant cause for the destabilization of the O/W emulsion during freezing and thawing is the crystallization of the oil phase and that the effects of the freezing and thawing rates on the stability are insignificant.     

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。     

Heat Gelation of Oil-in-Water Emulsions Stabilized by Whey Protein

The conditions under which a high volume fraction of oil can be trapped in whey protein gels were studied. Oil-in-water emulsions of whey protein and vegetable oil were subjected to heat treatment. Such emulsions, depending on their protein and oil content, on their pH and on the emulsification technique used, gelled or remained liquid. Homogenization was the major factor to achieve gelation and the firmness of heat-induced gels increased with increasing emulsion fineness and homogeneity. Emulsions with a high gelation capacity were characterized by a single droplet family of relatively narrow size distribution and a mean droplet diameter ranging from roughly 300–700 nanometers. The pH range suitable for gelation extended from 3.5–8.0.     

大豆分离蛋白-甜菊糖苷复合稳定剂制备纳米乳液

为适应食品工业对食品配料天然绿色、营养健康的追求,采用大豆分离蛋白（soy protein isolate,SPI）-甜菊糖苷（steviol glycosides,STE）复合体系作为稳定剂制备纳米乳液,研究稳定剂组成、微射流参数、油相质量分数等对纳米乳液形成的影响,并对乳液稳定性及微结构进行表征。结果表明：油相质量分数为10%时,单独SPI（1%）制备的乳液粒度较大（d43为0.548 μm）,稳定性差。添加0.25%～1% STE时,乳液粒度分布更均匀,粒度变小;当STE质量分数为0.5%和1%时,乳液粒度小于200 nm,且具备较好的贮存稳定性（30 d）。添加2% STE会导致乳滴表面蛋白被完全取代,从而弱化乳液的长期稳定性。微射流压力、均质次数及STE质量分数的增加均可降低乳液粒度,但油相质量分数的增加可增加乳液粒度。进一步将纳米乳液进行冷冻干燥处理,可制得结构化良好且高油含量的油粉;相对于单独SPI稳定的结构化乳液,SPI-STE纳米乳液制得的油粉结构更为完整,表面黏性小。     

乳清分离蛋白美拉德反应产物的体外抗氧化特性

以乳清分离蛋白和木糖、葡萄糖、果糖、蔗糖、麦芽糖、乳糖为美拉德反应原料,研究蛋白与糖的种类及混合比例对褐变程度的影响,并对反应产物进行抗氧化活性检测。结果表明:木糖与乳清分离蛋白按质量比2:1混合,湿热糖基化反应后,褐变程度最强,且在乳清分离蛋白与木糖按质量比2:1条件下反应后,乳清分离蛋白-木糖体系的美拉德反应产物(MRPs)具有较高还原能力和抗脂质过氧化能力及对DPPH自由基和O-2.具有较高的清除能力。     

酒精对酪蛋白酸钠溶液及O/W乳状液的影响

介绍了酒精对酪蛋白酸钠溶液及酪蛋白稳定的O/W乳状液性质的影响 .试验表明酒精在一定程度上可以降低酪蛋白酸钠的溶解度 .界面张力的测定则表明酒精的存在在很大程度上可以降低油—水界面和油—酪蛋白溶液界面的界面张力 .含酒精的乳状液体系的粘度会由于酒精的存在而提高 ,在酒精体积分数达 3 0 %时 ,乳状液体系的粘度会突然大幅度升高 .通过O/W乳状液的分层稳定性测定可发现 ,低浓度的酒精可以提高酪蛋白稳定的乳状液的分层稳定性 ,但酒精质体积分数超过 3 2 %时 ,乳状液的分层稳定性会受到破坏 .含酒精的O/W乳状液体系中油相含量的提高在一定范围内可以提高乳状液的稳定性 ,但高分散相浓度的含酒精的乳状液体系中由于连续相中酒精浓度的提高使乳状液体系稳定性下降 .     

一步法构建大豆蛋白双重乳液及其性质研究

本研究发现在中性条件下可以利用酸热处理的大豆分离蛋白（SPI）并通过简单的一步均质法制备得到W/O/W双重乳液,进一步研究了不同酸热处理时间对SPI在中性条件下的溶解度及其乳液性质（如乳析率、粒度）的变化。结果表明,不同时间的酸热处理会导致SPI在中性条件下的溶解度出现不同程度的下降（未加热SPI高达78.60%,酸热处理SPI为36.02%~53.29%）,可能是由于形成了不溶性的蛋白纤维聚集体,维系这些不溶聚集体的主要作用力是氢键和二硫键。酸热处理可以显著改善SPI稳定的乳液的乳析稳定性,且长时间的酸热处理效果更佳（如未加热SPI,38.70%;酸热处理2~6 h,34.90%~36.50%;酸热处理12~20 h,17.50%~19.70%）,但是酸热处理会导致乳滴粒度增大（如未加热SPI,24.63 μm;酸热处理SPI,28.12~33.97 μm）。增大乳液体系中的油含量也可以明显降低乳液的乳析率和粒度,随着油含量从20%增加到60%,乳析率从76.20%下降到3.30%,粒度从28.23 μm减小到19.06 μm。本研究结果为制备性质可控的双重乳液提供了新方法。     

Water/Oil Emulsions Prepared by the Membrane Emulsification Method and Their Stability

We developed and tested a simple method to measure dispersed droplet size of W/O emulsions. Then, using a microporous glass membrane treated with oil phase, we produced a W/O emulsion with high water content (40% w/w) at a high emulsification rate by the membrane emulsification method, and assessed its stability. In comparison with emulsions by the stirring methods, variations in dispersed droplet size and viscosity of emulsions by membrane method were small and the emulsions were more stable. Droplet size was not related to the stability of the W/O emulsion prepared by membrane emulsification.     

亲脂性乳化剂的性质和用量对W/O/W型复合乳状液稳定性的影响

研究了亲脂性表面活性剂的性质和用量对W/O/W型复合乳状液稳定性的影响,采用两种类型的亲脂性表面活性剂：新型的聚合型高分子表面活性剂聚氧乙烯（30）二聚羟基硬脂酸脂（Arlacel P135）和传统的小分子表面活性剂Span80和Tween80的混合物（质量比9∶1,HLB=5.4）制备复乳。采用离心保留率、光学显微镜和黏度等方法评价初乳和复乳的稳定性。结果表明,与小分子表面活性剂Span80和Tween80的混合物相比,大分子聚合表面活性剂Arlacel P135可以显著地提高复合乳状液的稳定性,在优化的条件下复乳的稳定性可以达到2000r/min离心15min不分层。