大豆分离蛋白对肌原纤维蛋白加热过程中结构及流变特性的影响

大豆分离蛋白（soybean protein isolate,SPI）作为优质的植物蛋白常被用于肉制品加工中,以提高产品产量和质地。研究添加SPI对肌原纤维蛋白（myofibrillar protein,MP）凝胶特性及MP加热过程中结构和流变特性的影响。结果表明：添加10%、20% SPI能提升混合凝胶的凝胶强度及保水性（P＜0.05）;加热过程中混合蛋白凝胶二级结构发生改变,但其变化规律尚不明确;添加SPI使混合凝胶储能模量及损耗模量下降;混合凝胶上清液十二烷基硫酸钠-聚丙烯酰胺凝胶电泳图谱显示,肌球蛋白重链、肌动蛋白、SPI部分亚基均是参与凝胶形成的蛋白质。     

碱性蛋白酶控制酶解诱导大豆分离蛋白纳米颗粒的形成机制

以大豆分离蛋白（soy protein isolate,SPI）为原料,利用碱性蛋白酶对其进行酶解处理（0～24 h）,探究SPI的结构变化规律,发现碱性蛋白酶控制酶解可诱导SPI自组装形成系列分布均匀（多相分散系数＜0.3）、粒径可控（90～200 nm）且具有不同表面特性的大豆蛋白纳米颗粒（soy protein nanoparticles,SPNs）,其中水解度（degree of hydrolysis,DH）及亚基解离/降解是影响SPNs形成的关键性因素。酶解初期（10～30 min,DH约3%）,SPI中β-伴大豆球蛋白（7S）组分α与α’亚基的部分降解有利于两亲性结构的释放,提高蛋白表面疏水性,降低临界聚集浓度,形成包含相对完整的7S及大豆球蛋白（11S）亚基的I类纳米颗粒（SPNs-DH 3%）。随着酶解时间的延长（1～2 h）,α与α’亚基的进一步降解促进了疏水性β亚基与B亚基的暴露,增强的疏水相互作用导致体系浊度增加,其中可溶性聚集体向不溶性疏水聚集的转化使得蛋白表面疏水性急剧下降,形成以A亚基及部分β亚基为主导的II类亲水型纳米颗粒（SPNs-DH 5%）。酶解后期（4～24 h）,A亚基的进一步降解则产生更多亲水性多肽,不利于纳米颗粒的形成。进而探究SPNs的形成机制,圆二色光谱结构表明,SPNs的形成与蛋白α-螺旋和无规卷曲结构向β-折叠转化有关。两类SPNs的整体结构均由疏水相互作用维持,而氢键和二硫键分别参与颗粒表面与内部结构的形成。与SPNs-DH 3%相比,SPNs-DH 5%中形成了更多由二硫键与氢键稳定的折叠结构。此外,由于酶解过程中不断释放抗氧化肽段,其所形成SPNs的抗氧化性较原始SPI均有所提升。     

亚基水平上大豆蛋白凝胶性的研究进展

大豆蛋白主要由β-伴大豆球蛋白（7S球蛋白）和大豆球蛋白（11S球蛋白）组成,不同亚基组成直接影响着大豆蛋白的加工特性。凝胶特性是本研究的主题。本文总结了大豆蛋白的组成及其化学结构,介绍7S球蛋白和11S球蛋白的凝胶形成过程,同时对亚基组分对凝胶性的影响和蛋白亚基缺失型大豆的研究进展进行概述,为大豆蛋白产品的开发及应用提供重要的理论指导。     

不同酸浆添加量制备的豆腐凝胶特性比较

以同一品种的大豆为原料制取豆浆后,添加不同比例酸浆（20%、23%、26%）,所得酸浆豆腐通过TPA质构、化学作用力、SDS-PAGE、微观结构、红外光谱等指标的测定,分析酸浆添加量对酸浆豆腐凝胶特性的影响。研究表明：随着酸浆添加量的增加,豆腐的质构性增强,在26%时豆腐的硬度达到最大值3.53 N,豆腐含水量、保水性及得率均显著下降（p<0.05）,蛋白质含量从13.09%增加到15.28%;二硫键和疏水相互作用是酸浆豆腐形成的主要作用力,二者占比超过90%,离子键和氢键的参与度相对较低。疏水作用是7S蛋白参与凝胶的主要作用力,11S蛋白的部分A亚基通过离子键和氢键连接形成凝胶,大分子蛋白聚集体、11S蛋白的B亚基和A3亚基主要通过二硫键参与凝胶的形成;随着酸浆添加量的增加,酸浆豆腐的凝胶网络结构逐渐变得粗糙,α螺旋含量由16.00%减少到15.83%,无规则卷曲由16.04%减少到15.80%,β转角由34.12%减少到33.57%,β折叠由33.84%增加到34.80%。综上可知,酸浆添加量对豆腐的凝胶特性有显著影响。     

外源蛋白对白鲢鱼糜肌原纤维蛋白结构及其结合特征腥味物质能力的影响

研究不同添加量（2%、4%、6%）大豆分离蛋白（soy protein isolate,SPI）、蛋清蛋白（egg white protein,EWP）、乳清分离蛋白（wheyproteinisolate,WPI）对白鲢鱼糜肌原纤维蛋白（myofibrillarprotein,MP）结构及其结合特征腥味物质（己醛、壬醛、1-辛烯-3-醇）能力的影响。结果表明,适量外源蛋白可促进MP展开,MP的表面疏水性、浊度、粒径、Zeta电位也随之增加,此时暴露出的疏水基团可增强MP结合己醛、壬醛的能力,暴露出的羟基则可增强MP对1-辛烯-3-醇的结合。外源蛋白添加量进一步增加,添加EWP、WPI的MP结合1-辛烯-3-醇的能力由于蛋白表面增加的亲水基团进一步增强。此外,添加WPI能更好地增强MP结合3种腥味物质的能力。     

超高压处理对乳清分离蛋白凝胶特性的影响

研究了超高压处理压力、时间、蛋白质量浓度、pH值、CaCl_2对乳清分离蛋白(whey protein isolated,WPI)凝胶特性的影响。当处理压力≥300 MPa、处理时间≥10 min、蛋白质质量浓度≥12 g/L时,WPI溶液经超高压处理后可以形成凝胶,且随着处理压力增大、处理时间延长和蛋白质浓度的提高,凝胶中二硫键含量明显升高,凝胶网络结构趋于致密,质地逐渐细腻,凝胶强度、得率和保水性呈现增大的趋势;WPI溶液pH在等电点以上且接近中性时,形成凝胶的二硫键含量较高,凝胶网络致密,凝胶品质较好;添加CaCl_2对形成凝胶的二硫键含量不产生影响,但其可以通过键桥作用提高凝胶强度;凝胶得率与CaCl_2浓度呈负相关。当CaCl_2浓度为0.06mol/L时,凝胶具有较大保水能力,之后保水性随Ca~(2+)浓度的升高而下降。     

超声处理对大豆分离蛋白-乳清分离蛋白混合蛋白功能特性的影响

为了研究超声处理对大豆分离蛋白（soy protein isolate,SPI）-乳清分离蛋白（whey protein isolate,WPI）混合体系乳化性、凝胶性以及结构状态的影响,采用不同超声功率处理混合蛋白体系,分析混合蛋白乳化活性、乳化稳定性、凝胶强度、持水性等功能特性变化,并采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、紫外光谱、扫描电子显微镜研究其结构特征变化。结果表明：SPI-WPI混合体系在超声功率为300 W时乳化活性与乳化稳定性分别达到最大值（76.46 m2/g和22.83 min）;紫外光谱发生轻微红移,说明内部基团暴露,蛋白结构发生改变;SPI-WPI混合体系凝胶强度与持水性在超声功率300 W时均达到最大值,分别为1 000.93 g和87.11%,与扫描电子显微镜观察结果一致,混合蛋白凝胶具有致密、规则的三维网状结构。说明超声处理能有效提高SPI-WPI混合蛋白的功能特性。     

富硒发芽对大豆蛋白凝胶性质的影响

以10 mg/L Na2SeO3溶液浸泡的大豆为材料,采用碱提酸沉法制备大豆分离蛋白(soybean protein isolate,SPI),以葡萄糖酸-δ-内酯(GDL)为凝固剂制备大豆蛋白凝胶,系统研究了富硒处理及发芽时长对大豆蛋白凝胶性质的影响。结果发现:发芽48 h内大豆GDL凝胶与SPI凝胶的硬度快速下降,分别由25.25 g和27.73 g降至10.77 g和13.37 g,持水性从61.42%和62.64%分别降至51.55%和55.54%。SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)图谱显示,富硒大豆与对照SPI谱带变化基本一致,其中7S球蛋白的β亚基与11S球蛋白的碱性亚基B较稳定,而7S球蛋白的α'亚基和α亚基与11S球蛋白的酸性亚基A3和A则被内源蛋白酶逐渐降解为相对分子质量较小的组成,从而影响发芽大豆凝胶性质。而富硒处理对发芽大豆蛋白凝胶性质影响较小。     

不同食用蛋白的添加对鲤鱼鱼糜流变和凝胶特性的影响

目的:研究大豆分离蛋白(soybean protein isolated,SPI)、乳清分离蛋白(whey protein isolate,WPI)、花生分离蛋白(peanut protein isolate,PPI)的添加对鲤鱼鱼糜流变和凝胶性质的影响。方法:利用流变仪、质构仪、色差计等对添加不同蛋白鱼糜的弹性模量、黏性模量、凝胶强度、破断强度、凹陷深度、持水性以及白度进行测定,并采用相关性分析法研究各指标之间的相互关系。结果:不同添加量的SPI、WPI和PPI均能有效地改善鱼糜的弹性模量、黏性模量、破断强度、凝胶强度和持水性,但会降低破断深度和白度,但各测定指标间存在显著相关(p<0.05)。SPI和PPI的添加对鱼糜的流变性、破断强度、凝胶强度的提高效果更好,添加量为8%时,鱼糜的凝胶强度均达到最大值,其中SPI组可达3806.70 g·mm,比对照组增加了34.63%;WPI对鱼糜的保水性效果最好,添加量为8%时,失水率仅为12.6%;白度随着蛋白添加量的增加而降低,其中PPI组与WPI组引起的白度降低较少,且差异不显著(p>0.05)。结论: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。     

乳清分离蛋白与变性木薯淀粉混合凝胶性质研究

研究了淀粉比例及木薯淀粉改性方式对乳清分离蛋白(WPI)与淀粉混合凝胶的影响。结果显示:当淀粉比例低于50%时,淀粉与WPI产生了协同效性,改善了混合凝胶储能模量;淀粉的羟丙基改性与交联改性均提高了混合凝胶G’;微观结构观察结果显示:淀粉膨胀颗粒被包裹在蛋白凝胶网络中,强化了凝胶结构;羟丙基淀粉在蛋白凝胶网络中较原淀粉更加破碎;而交联淀粉保留了较多完整的颗粒结构;WPI-交联淀粉混合凝胶呈现出完整淀粉颗粒周围附着蛋白胶粒的微观结构,强化了网络结构。     

Acid gelation properties of fibrillated model milk protein concentrate dispersions

Whey proteins in milk are globular proteins that can be converted into fibrils to enhance functional properties such gelation, emulsification, and foaming. A model fibrillated milk protein concentrate (MPC) was developed by mixing micellar casein concentrate (MCC) with fibrillated milk whey proteins. Similarly, a control model MPC was obtained by mixing MCC with milk whey proteins. The resulting fibrillated model MPC and control model MPC contained 5% protein and a ratio of casein to whey proteins similar to milk. The objective of the current study was to understand the rheological characteristics of fibrillated and control model MPC during acid gelation, using Förster resonance energy transfer (FRET) to assess small amplitude oscillation and casein–whey protein interaction. The results from the FRET index images showed greater interactions between caseins and whey proteins in fibrillated model MPC compared with the moderate and uniform interactions in control model MPC gels. Rheological study showed that the maximum storage modulus of acid gel of fibrillated model MPC was 546.9 ± 15.5 Pa, which was significantly higher than acid gel made from control model MPC (336.9 ± 11.3 Pa), indicating that fibrillated model MPC produced a firmer gel. Therefore, it can be concluded that acid gel produced from fibrillated model MPC was stronger than control model MPC. Selective fibrillation of the whey protein fraction in MPC can be used to improve gelation characteristics of acid gel type products.     

Modification of rheological properties of whey protein isolates by limited proteolysis

Whey protein isolate (WPI) was subjected to limited tryptic hydrolysis and the effect of the limited hydrolysis on the rheological properties of WPI was examined and compared with those of untreated WPI. At 10% concentration (w/v in 50 mM TES buffer, pH 7.0, containing 50 mM NaCl), both WPI and the enzyme-treated WPI (EWPI) formed heat-induced viscoelastic gels. However, EWPI formed weaker gels (lower storage modulus) than WPI gels. Moreover, a lower gelation point (77 °C) was obtained for EWPI as compared with that of WPI which gelled at 80 °C only after holding 1.4 min. Thermal analysis and aggregation studies indicated that limited proteolysis resulted in changes in the denaturation and aggregation properties. As a consequenece, EWPI formed particulated gels, while WPI formed fine-stranded gels. In keeping with the formation of a particulate gel, Texture Profile Analysis (TPA) of the heat-induced gels (at 80 °C for 30 min) revealed that EWPI gels possessed significantly higher (p < 0.05) cohesiveness, hardness, gumminess, and chewiness but did not fracture at 75% deformation. The results suggest that the domain peptides, especially β-lactoglobulin domains released by the limited proteolysis, were responsible for the altered gelation properties.     

Addition of sodium caseinate to skim milk increases nonsedimentable casein and causes significant changes in rennet-induced gelation,heat stability,and ethanol stability

The protein content of skim milk was increased from 3.3 to 4.1% (wt/wt) by the addition of a blend of skim milk powder and sodium caseinate (NaCas), in which the weight ratio of skim milk powder to NaCas was varied from 0.8:0.0 to 0.0:0.8. Addition of NaCas increased the levels of nonsedimentable casein (from ～6 to 18% of total casein) and calcium (from ～36 to 43% of total calcium) and reduced the turbidity of the fortified milk, to a degree depending on level of NaCas added. Rennet gelation was adversely affected by the addition of NaCas at 0.2% (wt/wt) and completely inhibited at NaCas ≥0.4% (wt/wt). Rennet-induced hydrolysis was not affected by added NaCas. The proportion of total casein that was nonsedimentable on centrifugation (3,000 × g, 1 h, 25°C) of the rennet-treated milk after incubation for 1 h at 31°C increased significantly on addition of NaCas at ≥0.4% (wt/wt). Heat stability in the pH range 6.7 to 7.2 and ethanol stability at pH 6.4 were enhanced by the addition of NaCas. It is suggested that the negative effect of NaCas on rennet gelation is due to the increase in nonsedimentable casein, which upon hydrolysis by chymosin forms into small nonsedimentable particles that physically come between, and impede the aggregation of, rennet-altered para-casein micelles, and thereby inhibit the development of a gel network.     

Effect of CMC Molecular Weight on Acid‐Induced Gelation of Heated WPI‐CMC Soluble Complex

Acid‐induced gelation properties of heated whey protein isolate (WPI) and carboxymethylcellulose (CMC) soluble complex were investigated as a function of CMC molecular weight (270, 680, and 750 kDa) and concentrations (0% to 0.125%). Heated WPI‐CMC soluble complex with 6% protein was made by heating biopolymers together at pH 7.0 and 85 °C for 30 min and diluted to 5% protein before acid‐induced gelation. Acid‐induced gel formed from heated WPI‐CMC complexes exhibited increased hardness and decreased water holding capacity with increasing CMC concentrations but gel strength decreased at higher CMC content. The highest gel strength was observed with CMC 750 k at 0.05%. Gels with low CMC concentration showed homogenous microstructure which was independent of CMC molecular weight, while increasing CMC concentration led to microphase separation with higher CMC molecular weight showing more extensive phase separation. When heated WPI‐CMC complexes were prepared at 9% protein the acid gels showed improved gel hardness and water holding capacity, which was supported by the more interconnected protein network with less porosity when compared to complexes heated at 6% protein. It is concluded that protein concentration and biopolymer ratio during complex formation are the major factors affecting gel properties while the effect of CMC molecular weight was less significant.     

Acid-induced gelation of milk protein concentrates with added pectin: Effect of casein micelle dissociation

The dynamics of the formation of the acid gel network for mixtures of milk protein concentrate (MPC) and low methoxyl amidated (LMA) pectin were studied using rheological measurements. The results as a function of pectin content and casein micelle integrity, from neutral pH to approximately pH 4.2, together with the microstructural changes observed in some of these systems, are presented.The gelation profiles of a mixture of 4% w/v MPC and LMA pectin (0–0.075% w/v) after the addition of 1.2% w/v glucono-δ-lactone showed a gradual decrease in the shear modulus with the incorporation of pectin. The effects of casein micelle integrity on casein–pectin interactions were studied, by preparing MPC dispersions containing various levels of micellar casein. A gradual change in the shear modulus, from a disrupting effect of pectin added to MPC, in which the casein micelles are intact, to a clear synergistic effect of pectin added to dissociated casein systems, was found in the acid-induced milk gels.     

Assessment of the effects of soy protein isolates with different protein compositions on gluten thermosetting gelation

Although soy proteins are known to have a deleterious effect on gluten thermosetting gelation, the causes are still poorly understood. Different sources of soy protein isolates (SPI) were used to investigate the interactions between gluten and soy proteins during hydro-thermal treatments. Commercial SPI and isolates prepared from soybean lines with different subunit composition were used to study the influence of protein denaturation and subunit composition on thermoset gel formation. Rapid Visco Analyser analysis showed that replacement of gluten with more than 1% SPI decreased the peak viscosity and interfered with formation of thermoset gels. However, peak viscosity was higher for 11% gluten + 2% SPI than for 11% gluten alone, suggesting a cooperative effect. After heating and cooling, 11% gluten + 2% SPI rich in A1 and A2 subunits formed a coherent thermoset gel suggesting that the cysteine residue content of soy proteins can affect gel formation.     

Combined acid‐ and rennet‐induced gelation of a mixed soya milk–cow's milk system

The objective of this study was to better understand the gelation behaviour of a mixed soya milk–cow's milk system, by forming different reactive protein particles using rennet and glucono‐δ‐lactone. The formation of the structure of these mixed gels was followed, for the first time, using diffusing wave spectroscopy and rheology. When only one protein source was induced to gel, protein aggregation was hindered, as shown by the slower increase in apparent radius after the gel point. Confocal microscopy analysis of the gel networks suggested that while milk gels exhibited large pores with interconnecting strands of protein, soya gels appeared as densely packed protein aggregates, and mixed soya milk gels appeared as a network of aggregated proteins. This study demonstrated that by modulating the reactivity of the building blocks, it is possible to fine‐tune structure formation of these mixed protein gels.     

Prevention of low-temperature gelation in milk protein concentrates by calcium-binding salts

The objective of this study was to determine the effect of adding low concentrations of calcium-binding salts on the prevention of low-temperature gelation in milk protein concentrates (MPC). The MPC were created by a combination of ultrafiltration and diafiltration, standardized from 14 to 17% (wt/vol) protein content and mixed with one of 5 calcium-binding salts (sodium citrate, sodium hexametaphosphate, sodium polyphosphate, sodium pyrophosphate, and sodium monophosphate) adjusted to a pH of 6.75. The flow properties, apparent viscosity, and gel strength were determined for MPC containing a wide range of calcium-binding salt concentrations. Low-temperature gelation occurred in MPC with 16.0% and higher protein content. Low-temperature gelation at 16.0% protein content was prevented by the addition of any of the 5 salts tested at low concentrations (0.30 mM or less; sodium citrate, sodium hexametaphosphate, sodium polyphosphate, sodium pyrophosphate or sodium monophosphate), with sodium polyphosphate and sodium monophosphate being the most consistent in preventing low-temperature gels. All MPC samples exhibited shear-thinning behavior (n = 0.52–0.72), which increased (lower n values) as the protein content increased and decreased by addition of salt. At concentrations of salt above 1.00 mM, thermally irreversible gels were observed with relative strength dependent on the salt and protein content.     

Effects of disulphide bonds between added whey protein aggregates and other milk components on the rheological properties of acidified milk model systems

Micro- and nanoparticulated whey protein (MWP, NWP) were added to non-fat milk model systems and processed into chemically (glucono-delta-lactone) acidified milk gels. Model systems contained 5% protein in total and were made at two levels of casein (2.5% and 3.5%, w/w) with and without the thiol-blocking agent N-ethylmaleimide. The systems were characterised in terms of thiol groups, gel electrophoresis, particle size, and rheology during processing (homogenisation, heat treatment and acidification). The results showed that the formation of disulphide-linked structures in milk model systems was closely related to the increased particle size and rheological behaviour of the gels. MWP enriched systems produced, upon acidification, weak protein networks and required the addition of whey protein isolate (WPI) to increase gel strength. However, systems containing NWP exhibited pronounced increase in particle size and higher firmness of acidified gels through both covalent and non-covalent interactions.     

Heat Stability of Oil-in-Water Emulsions Containing Milk Proteins: Effect of Ionic Strength and pH

Emulsions (20 wt% soybean oil; 2 wt% protein) made with caseinate at pH 7 and with whey protein isolate (WPI) at pH 7 and 3 were stable to heating at 90 and 121°C. WPI emulsions destabilized at pH values between 3.5 and 4.0. In the presence of KCI (12.5–200 mM), large particles were formed in WPI emulsions at pH 3 and the emulsions were viscous. At pH 7, moderate concentrations of KCI decreased the heat stability and gels were formed. KCI had less effect on WPI emulsions made at pH 3. Combining the emulsions with caseinate allowed some control of the heat-induced gelation.