过氧自由基氧化对大米蛋白结构的影响

采用不同浓度2,2’-盐酸脒基丙烷(AAPH)在有氧条件下热分解产生的过氧自由基代表脂质氧化过程中的脂质自由基,研究过氧自由基氧化对大米蛋白结构的影响。结果表明,当AAPH浓度从0增加至25 mmol/L,大米蛋白羰基和二硫键含量分别由3.77和13.88 nmol/mg增加至7.26和15.73 nmol/mg,游离巯基由7.32 nmol/mg下降至1.97 nmol/mg,表明过氧自由基诱使大米蛋白发生了氧化;傅里叶红外分析表明蛋白质氧化导致大米蛋白α-螺旋和β-折叠含量下降,β-转角和无规卷曲含量上升。随着大米蛋白氧化程度的增加,大米蛋白粒径从126 nm增加到216 nm,表面疏水性从1053下降到568,内源荧光最大荧光峰位蓝移,内源荧光强度下降,并且在分子量分布图中高分子量聚集体含量逐渐增加。表明过氧自由基氧化修饰导致大米蛋白结构改变和形成氧化聚集体。     

利用酪蛋白酸钠的分子伴侣特性抑制蛋清蛋白的热聚集研究

酪蛋白酸钠的分子伴侣作用尚未在蛋清蛋白体系的热聚集研究中应用。通过测定溶液的浊度、上清液总蛋白含量、粒径分布以及分子质量分布变化,研究酪蛋白酸钠添加量对蛋清蛋白热聚集的影响。当酪蛋白酸钠质量浓度为2mg/mL时可以抑制热处理(90℃,30min)蛋清蛋白溶液(5mg/mL)的热聚集,且随着酪蛋白酸钠添加量的增加(2~3mg/mL),其抑制程度增加;当酪蛋白酸钠质量浓度较高时(4~5mg/mL),由于酪蛋白酸钠的溶解性较低,抑制效果无显著变化(P<0.05)。还原SDS-PAGE电泳结果表明,酪蛋白酸钠对蛋清蛋白中的卵白蛋白、卵转铁蛋白热聚集的抑制效果决定了其对蛋清蛋白热聚集抑制效果。随着酪蛋白酸钠添加量的增加,热处理蛋清蛋白溶液中的不溶性聚集逐渐转变为可溶性聚集,结构紧密且体积较大的热聚集体转变为结构疏松体积较小的聚集体。研究结果表明,酪蛋白酸钠的添加可以抑制蛋清蛋白的热聚集。     

不同离子强度大豆分离蛋白热诱导聚集体的研究

目的:研究离子强度对大豆分离蛋白热诱导聚集的影响.方法:采用可见分光光度计,在波长600 nm处测定不同离子强度下的大豆分离蛋白热处理溶液的吸光值,并将其作为溶液的浊度;采用体积排阻色谱(SECHPLC)、动态激光光散射(DLS)及zeta电位仪研究不同离子强度下大豆分离蛋白热诱导聚集体的分子质量分布、粒径分布及zeta电位.结果:随着离子强度的增加,大豆分离蛋白聚集体溶液的浊度增加,体系的聚集体部分及平均流体动力学半径(Rh)大幅增加,zeta电位逐渐降低.结论:电荷屏蔽作用使分子间斥力降低,促进了高离子强度下聚集的产生.     

热处理对卵白蛋白理化性质及其ACE抑制活性的影响

在不同温度下对卵白蛋白的粒径、Zeta电位、巯基含量、内源荧光、表面疏水性等理化性质以及酶解后卵白蛋白的水解度、血管紧张素转化酶(angiotensin converting enzyme,ACE)抑制活性进行研究。结果表明:随着热处理温度的增加,卵白蛋白粒径分布发生了变化,平均粒径先增大后减小,从(622.73±4.60)nm增大到(1 527.00±59.40)nm,然后减小至(261.64±2.26)nm;Zeta电位绝对值从(6.56±0.36)m V增大到(19.20±0.62)mV;表面巯基含量从(2.20±0.37)μmol/g增加到(23.76±0.71)μmol/g,总巯基含量则先减少后增大,从(9.63±0.26)μmol/g减少到(6.46±0.16)μmol/g,然后增至(29.55±1.88)μmol/g;内源荧光最大吸收峰发生1 nm的蓝移;表面疏水性值先增大后减小,从(249.52±0.49)增大至(274.32±1.19),然后减小至(135.17±14.81);水解度先增大后减小,60℃时可达到(28.62±0.42)%;ACE抑制活性有着不同程度的增大,80℃时的ACE抑制活性达到(79.17±0.54)%,比未处理溶液的ACE抑制活性高了23.01%。     

蛋白质氧化对核桃蛋白质结构的影响

为了深入明晰氧化对核桃蛋白质结构性的影响,本文以核桃分离蛋白为研究对象,采用不同浓度的2,2'-盐酸脒基丙烷(AAPH)热降解形成烷过氧自由基(ROO·)代表脂质过氧化反应过程中产生的脂质自由基,对核桃分离蛋白进行氧化修饰得到不同氧化程度的核桃氧化蛋白。对不同氧化程度蛋白的羰基、巯基和总巯基,表面疏水性(H₀)、内源荧光、粒径分布、相对分子质量和二级结构进行研究和分析。结果表明,随着AAPH浓度的增加,核桃氧化蛋白羰基含量显著增加(p<0.05),巯基和H₀显著降低(p<0.05);圆二色谱结果表明,随着AAPH浓度的增加,核桃蛋白α-螺旋结构和β-折叠含量下降,而无规卷曲结构含量增加,说明氧化破坏了蛋白质的二级结构,使有序结构变为无序结构。最大内源荧光值降低,且出现蓝移现象,说明烷过氧自由基氧化导致分子聚集,使得色氨酸残基包埋;粒径分布结果表明,核桃氧化蛋白可溶性聚集体可被共价交联和非共价聚集转变成不可溶性部分;体积排阻色谱结果表明,核桃氧化蛋白相对分子质量随着氧化浓度的增大聚集程度逐渐增大。研究结果表明,核桃蛋白在烷过氧自由基(ROO·)氧化体系中产生了显著氧化作用并导致其结构发生一定的改变,为进一步阐明核桃蛋白氧化机理提供理论依据。     

米糠贮藏过程中清蛋白结构变化的研究

以不同贮藏时间新鲜米糠为原料脱脂制备米糠清蛋白,研究新鲜米糠贮藏时间对米糠清蛋白结构的影响。结果表明:新鲜米糠制备米糠清蛋白的羰基、游离巯基和二硫键含量分别为1.55、46.56、4.01 nmol/mg,随着新鲜米糠贮藏时间的延长,米糠清蛋白的羰基和二硫键含量增加,游离巯基含量下降,新鲜米糠贮藏10 d制备米糠清蛋白的羰基、游离巯基和二硫键含量分别为6.55、29.40、7.87 nmol/mg,表明新鲜米糠贮藏过程中米糠清蛋白发生了氧化;当新鲜米糠贮藏时间从0d增加到10 d,米糠清蛋白粒径从60.18 nm逐渐增加到82.69 nm,溶解性从88.54%逐渐下降到56.00%,表面疏水性指数从789.56逐渐下降到608.97,Zeta电位绝对值从12.26 mV下降到8.83mV,表明蛋白质氧化导致米糠清蛋白形成氧化聚集体;电泳分析表明非二硫共价键参与米糠清蛋白氧化聚集体的形成。     

超声波辅助碱处理增溶米渣蛋白工艺优化

采用超声波辅助碱处理提高米渣蛋白的溶解度。选择反应温度、超声波时间、超声波强度、蛋白浓度、Na OH浓度为优化因素,通过单因素和响应面分析,得到超声波辅助碱处理增溶米渣蛋白最佳工艺条件为:反应温度50℃、超声时间60 min、超声波强度19.2 W/cm~2、米渣蛋白浓度5.1%(w/v)、Na OH浓度0.08 mol/L。在此条件下,改性后得到的米渣蛋白溶解度达(20.09±0.58)mg/m L(w/v)。SDS-PAGE结果表明蛋白的二硫键和亚基遭到破坏,处理过程中伴随着一些不溶性蛋白聚集体的溶解,且蛋白平均粒径由485 nm降低到223 nm,进而导致米渣蛋白溶解度显著增加。这些结果表明超声波辅助碱处理有助于进一步加工利用米渣蛋白,为食品的生产加工提供借鉴。     

大豆多糖对乳清分离蛋白—乳状液稳定性与流变特性的影响

本文研究了大豆多糖(SSPS)与乳清分离蛋白(WPI)乳状液静电组装,形成乳状液聚集体,考察了不同浓度的SSPS对WPI-乳状液稳定性与流变特性的影响,以期提高体系的粘弹性,形成高流变特性的食品体系。将不同浓度的大豆多糖与2%乳清分离蛋白乳状液(油相为20%)静电组装,分析乳状液的粒径,Zeta-电位,稳定性指数,流变性质和微观结构。结合剪切流变与微流变技术,深入研究了SSPS对乳清分离蛋白(WPI)乳状液流体特性与结构的影响。结果表明:随着SSPS浓度的增加,WPI乳状液的粒径在添加0.25%SSPS时达到峰值(3350±0.35)nm,而后随着SSPS浓度的增加而降低;Zeta-电位绝对值呈递减的趋势,表明SSPS与WPI间产生静电吸附作用;SSPS静电吸附提高WPI乳状液的稳定性;剪切流变结果表明,SSPS浓度为0.5%时,其粘度最大,并在剪切速率为95.8 s-1处其粘度是WPI乳状液粘度的10倍以上;微流变结果表明,0.5%SSPS-WPI乳状液的MSD曲线出现平台区,表明其弹性指数(EI)与宏观粘度指数(MVI)均显著提高达到最大值。微观结构结果表明,0.5%SSPS-WPI乳状液形成均一的乳状液聚集体。本研究将有助于理解大豆多糖与蛋白质乳状液的相互作用,同时为低脂高流变特性的食品(如蛋黄酱、调味汁、巧克力和植脂奶油等)生产提供理论指导。     

pH偏移促进大豆蛋白酶解过程中聚集体解聚的研究

酶法水解大豆蛋白可以生产多种生物活性肽,但在酶解过程中肽段的二次聚集会产生不溶沉淀,降低蛋白质利用率。研究了pH偏移对大豆蛋白酶解聚集体解聚的影响。结果表明：Alcalase碱性蛋白酶酶解大豆蛋白获得的不溶沉淀,是以疏水相互作用为主要分子间作用力的大豆肽聚集体;pH偏移赋予的静电斥力能促使大豆肽聚集体解聚和重排,大豆肽蛋白质利用率从74%（pH 7）增加到85%（pH 12）,并将大豆肽的平均粒径由243 nm减小至103 nm;同时,pH偏移使疏水性氨基酸在不溶肽聚集体中积累,增加二级结构中的无规则卷曲含量,但不影响不溶肽聚集体的相对分子质量分布。因此,pH偏移技术可安全有效地促使大豆肽不溶聚集体解聚。     

紫苏油乳液的制备及其稳定性影响因素研究

本研究针对紫苏油不饱和脂肪酸含量高,在储存过程中易氧化等特点,采用高压均质法制备紫苏油乳液,通过激光粒度仪分析乳液粒径大小与分布,通过TURBISCAN浓缩体系稳定性分析仪监测乳液稳定性的变化趋势,探究乳化剂用量、油水比例、高压均质的压力和循环次数及HLB值对紫苏油乳液稳定性的影响,以提供一种紫苏油缓释方法,拓宽紫苏油在食品中的应用范围。试验结果表明,紫苏油乳液粒径主要分布在300~670nm;乳化剂浓度由0.2%增加至1.2%,乳液粒径下降,稳定性提高;浓度为1.2%时,乳液平均粒径(d=513nm)最小。随着油水比增加,紫苏油乳液稳定性下降;高压均质过程对乳液的稳定性有显著影响,压力越大,循环次数越高,乳液越稳定。与单一乳化剂(HLB=15)相比,复配乳化剂(HLB=8~14)可制得更为稳定的乳液,且当HLB值为11时,紫苏油乳液的平均粒径(d=374nm)最小,乳液稳定性最佳。     

Effect of concentration,ionic strength and freeze-drying on the heat-induced aggregation of soy proteins

The effects of protein concentration, ionic strength, and lyophilization on heat-induced aggregation of soy proteins were analyzed by SDS-PAGE, SEC-HPLC and laser light scattering (LLS). SDS-PAGE profile suggested that the aggregates were formed via non-covalent forces and/or disulfide bonds. At ionic strength of zero, SEC-HPLC revealed that the samples were composed of three major fractions: aggregates, intermediate fractions and non-aggregated molecules. Furthermore, the relative proportion of the aggregate fraction increased as protein concentration increased. Similarly, LLS indicated that the average hydrodynamic radius (R_h) increased at higher protein concentration. In sample with an ionic strength of zero, the intermediate fraction decreased after freeze-drying with a concomitant increase of the aggregate fraction. When the sample was heated at elevated ionic strength, the SEC-HPLC and LLS profiles changed substantially, the intermediate fractions decreased, and lyophilization had effect on the fraction of aggregates. These experiments suggest novel strategies for producing soy protein aggregates with controlled properties.     

大豆蛋白纤维聚集体的体外消化特性研究

本研究以大豆分离蛋白为材料,通过在pH 2.0条件下调控大豆蛋白形成大豆蛋白纤维聚集体。采用标准的静态体外消化模型INFOGEST 2.0对大豆蛋白纤维聚集体进行体外模拟消化实验,并对不同消化时间的胃肠消化产物通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）、动态激光散射仪（DLS）及液相色谱-质谱联用（LC-MS）对其分子质量、粒径及肽段组成进行鉴定。结果表明,大豆蛋白纤维聚集体通过消化后被水解为小分子的肽,并LC-MS证实消化过程中肽段数量随着消化时间的增加呈阶梯式水解。本实验探究了大豆蛋白在体外模拟消化过程中的消化特性,这将有助于更好地了解植物蛋白在人体中的消化方式及其对健康的影响。     

Effects of oxidative modification on thermal aggregation and gel properties of soy protein by peroxyl radicals

Effects of protein oxidation on thermal aggregation and gel properties of soy protein by 2,2′‐azobis (2‐amidinopropane) dihydrochloride (AAPH)‐derived peroxyl radicals were investigated in this article. Incubation of soy protein to increase concentration of AAPH resulted in a decrease in particle size and content of thermal aggregates during thermal‐induced denaturation. Protein oxidation resulted in a decrease in water‐holding capacity (WHC), gel hardness and gel strength of soy protein gel. An increase in coarseness and interstice of the gel network was accompanied by uneven distribution of interstice as extent of oxidation of soy protein increased. A decrease in disulphide content and formation of oxidation aggregates in the process of oxidative modification were contributed to the decline of particle size and content of thermal aggregates during thermal‐induced denaturation, leading to a decrease in WHC, gel hardness and gel strength of soy protein gel.     

大豆分离蛋白对大豆肽纳米颗粒Pickering乳液性能的影响

为提高大豆肽纳米颗粒(SPN)Pickering乳液稳定性,以大豆肽聚集体为原料,采用超声法制备SPN,对超声时间进行了优化;在SPN体系中引入大豆分离蛋白(SPI)构建复合乳化剂,研究不同乳化剂质量浓度下SPI对SPN界面活性和乳化稳定性的影响。结果表明：选取超声时间10 min制备SPN;随着乳化剂质量浓度的增大,乳液粒径逐渐减小,当乳化剂质量浓度较低(5 mg/mL)时,乳液出现桥联,乳化剂质量浓度过高(30 mg/mL)时则出现絮凝;界面蛋白吸附率随着乳化剂质量浓度的增加呈现先升高后降低的趋势。在相同乳化剂质量浓度下,添加SPI的SPN乳液(SPI-SPN乳液)的粒径分布峰左移,其粒径、界面蛋白吸附率显著小于SPN乳液的;在储存过程中,SPN乳液粒径逐渐增大,SPI-SPN乳液粒径没有显著变化;SPI-SPN乳液的乳析指数小于相同乳化剂质量浓度的SPN乳液,当乳化剂质量浓度为30 mg/mL时,储存15 d SPI-SPN乳液未出现分层现象。综上,SPI可以提高SPN的界面活性和SPN乳液储存过程中的絮凝稳定性和分层稳定性。     

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。     

Soybean protein aggregation induced by lipoxygenase catalyzed linoleic acid oxidation

Aggregation of lipid-reduced soybean proteins (LRSP) was investigated by chemical analysis, spectroscopy, electrophoresis, SEC-HPLC and light scattering. Soybean proteins obtained from the model systems consisting of LRSP and different levels of linoleic acid and lipoxygenase (RSP 4 and 5) showed increased turbidity, protein oxidation, surface hydrophobicity but decreased sulfhydryl and disulfide contents. SDS-PAGE of RSP 4 and 5 revealed remarkable difference of electrophoretic bands for 7S subunits, comparing with those samples without linoleic acid and lipoxygenase. Fluorescence spectroscopy suggested other covalent linkages than disulfide bonds formed during the formation of aggregates. SEC-HPLC and laser light scattering indicated that aggregates with high molecular weight and large particle size existed in samples of RSP 4 and 5. The experimental evidences suggest that the aggregates were formed via non-covalent interactions, but covalent bonds were also involved.     

百里香酚纳米复合诱导大豆蛋白自组装凝胶及机制

为了揭示百里香酚纳米复合诱导大豆蛋白自组装凝胶的途径及机制,该文研究了蛋白浓度、百里香酚添加量、反应初始pH值和反应终止pH值对大豆蛋白与百里香酚相互作用及其凝胶形成的影响。结果表明：四者协同调控大豆蛋白-百里香酚纳米复合物凝胶的形成,百里香酚荷载量的提高可降低大豆蛋白凝胶的临界蛋白浓度（75 mg/mL）,反应结束后调节pH值至中性是凝胶形成的必要条件。在pH值为7.0条件下,百里香酚复合诱导大豆蛋白产生了明显的纤维化聚集,其颗粒平均粒径增加27.60%、光散射强度提高154.67%、Th T荧光强度提高93.78%,因此推测百里香酚复合主要通过诱导大豆蛋白纤维化聚集的产生继而诱导凝胶的形成。大豆蛋白-百里香酚纳米复合物凝胶在尿素和DTT溶液中的溶解度提高,因而推测维持凝胶网络的分子间相互作用力主要为氢键、疏水相互作用力和二硫键。研究结果为大豆蛋白冷致凝胶提供了新的技术解决方案,并为百里香酚的开发利用提供理论支撑。     

Characterization of Oil-in-Water Emulsions Prepared with Commercial Soy Protein Concentrate

ABSTRACT: Oil-in-water emulsions containing a commercial fraction of soy protein concentrate (SPC) were characterized for stability and microstructural differences. Emulsions were prepared with SPC (concentrations between 1% and 10% (w/v)) and soybean oil (10% w/w) and homogenized at 80 MPa. When SPC was added at a concentration high enough to be present in the dispersed phase, the average particle size, as determined by integrated light scattering, reached a plateau value. In addition, emulsions prepared with > 4% SPC showed increased viscosity with increasing SPC concentration. The protein formed a continuous network and emulsions were stable to creaming. Microstructural observations showed that phase separation occurred in emulsions prepared with high SPC concentrations.     

Surface Properties of Heat‐Induced Soluble Soy Protein Aggregates of Different Molecular Masses

Suspensions (2% and 5%, w/v) of soy protein isolate (SPI) were heated at 80, 90, or 100 °C for different time periods to produce soluble aggregates of different molecular sizes to investigate the relationship between particle size and surface properties (emulsions and foams). Soluble aggregates generated in these model systems were characterized by gel permeation chromatography and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. Heat treatment increased surface hydrophobicity, induced SPI aggregation via hydrophobic interaction and disulfide bonds, and formed soluble aggregates of different sizes. Heating of 5% SPI always promoted large‐size aggregate (LA; >1000 kDa) formation irrespective of temperature, whereas the aggregate size distribution in 2% SPI was temperature dependent: the LA fraction progressively rose with temperature (80→90→100 °C), corresponding to the attenuation of medium‐size aggregates (MA; 670 to 1000 kDa) initially abundant at 80 °C. Heated SPI with abundant LA (>50%) promoted foam stability. LA also exhibited excellent emulsifying activity and stabilized emulsions by promoting the formation of small oil droplets covered with a thick interfacial protein layer. However, despite a similar influence on emulsion stability, MA enhanced foaming capacity but were less capable of stabilizing emulsions than LA. The functionality variation between heated SPI samples is clearly related to the distribution of aggregates that differ in molecular size and surface activity. The findings may encourage further research to develop functional SPI aggregates for various commercial applications.