Interactions and gel strength of mixed myofibrillar with soy protein, 7S globulin and enzyme-hydrolyzed soy proteins

The mixed protein gels were prepared adding soy protein isolate (SPI), 7S globulin, enzyme-hydrolyzed soy proteins, 10- to 100-kDa ultrafiltration fraction and 0.5- to 10-kDa ultrafiltration fraction to myofibril protein isolate (MPI) gels, and five chemical interactions namely nonspecific associations, ionic bonds, hydrogen bonds, hydrophobic interactions and disulfide bonds in these gels were investigated by means of determining gel solubility within 20–75 °C. Furthermore, correlations between gel strength and different chemical interactions were evaluated statistically by Pearson’s correlation test. The gels with 0.5- to 10-kDa fraction presented the biggest gel strength below 60 °C, and the gels with SPI had better gel strength above 65 °C. At different endpoint temperatures, nonspecific associations decreased in order of MPI mixed with 0.5- to 10-kDa fraction, 10- to 100-kDa fraction, enzyme-hydrolyzed soy proteins, 7S globulin and SPI. Gels with ultrafiltration fractions had higher ionic bonds. Hydrogen bonds fluctuated in small scale below 55 °C and reduced at higher temperature. Hydrophobic interactions increased to maximum before decreasing slowly as the temperature went on. In short, both hydrophobic interactions and ionic bonds had significantly positive correlation with gel strength for mixed gels with enzyme-hydrolyzed soy proteins, whereas for the other four mixed gels, it was hydrophobic interactions and nonspecific associations.     

Gelation Property of Alcohol-Extracted Soy Protein Isolate and Effects of Various Reagents on the Firmness of Heat-Induced Gels

Gelation property, thermal property, protein subunits distribution, and molecular forces involved in gelation of alcohol-extracted soy protein isolate were investigated using texture analyzer, differential scanning calorimeter, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and various reagents. Results showed that salts and pH played important roles in gel firmness, and a power law relationship between gel firmness and protein concentration was observed. The effects of various reagents revealed that disulfide bonds play a major role in soy protein gel formation, while the involvement of electrostatic interactions, hydrogen bonds, and/or hydrophobic interactions also occurred in gel networks. Thermal analysis indicated that both alcohol-extracted soy protein isolate and commercial soy protein isolate (isoelectric precipitation) have undergone serious denaturation, while the gel firmness of alcohol-extracted soy protein isolate was significantly greater than that of commercial soy protein isolate. Sodium dodecyl sulfate polyacrylamide gel electrophoresis image showed that there was almost no difference for protein subunits among alcohol-extracted soy protein isolate, commercial soy protein isolate, and soy powder. Hence, as an alternative environmental friendly extraction method, alcohol-extraction of soy protein isolate has a great prospect to replace presently applied isoelectric precipitation method.     

Papain-induced Gelation of Soy Glycinin (11S)

ABSTRACT:  The gelation of soy peptides produced by the action of papain enzymes on soy glycinin (11S) dispersions (4.7% w/v) was investigated. Cation-exchange chromatography was used to fractionate crude papain. The nonbinding fraction showed no gel-forming activity on the 11S dispersion. Two binding fractions showed gel-forming activity, and the gel strength of both 11S gels was similar. The activity of the crude papain on 11S dispersions produced a slightly stronger gel than one formed with either of the 2 binding fractions. With the crude papain, the rate of gel formation appeared to be strongly influenced by the enzyme concentration, but the maximum gel strength was independent of enzyme concentration. When the temperature was increased, the papain treatment of 11S soy protein produced weaker gels when the measurement was made at the temperature of formation. This dependence of maximum gel strength on temperature was found to be a function of only the measurement temperature and not the gel formation temperature. The degree of protein hydrolysis at maximum gel strength was similar (∼6%) for the gels formed at different temperatures. When the temperature was increased, the elastic modulus G', the viscous modulus G", and the degree of viscoelasticity (G"/G') decreased. This suggested that the gels were formed the by hydrophobic interactions among the peptides. This observation was supported by particle size measurements on samples of gels which were mixed with reagents known for their ability to disrupt hydrophilic/electrostatic, hydrophobic, or disulfide interactions.     

Binding of Alcohols by Soy Protein in Aqueous Solutions

Interactions of alcohols with soy protein isolate were investigated using an equilibrium dialysis method. It was found that interactions may involve hydrophobic association and, to some degree, hydrogen bonding. Studies with soy protein with various levels of denaturation indicated that denaturation of the protein by heating may limit its ability for the formation of hydrogen bonds with alcohols. The proteins under investigation exhibited practically unlimited binding capacity for alcohols.     

发芽对大豆蛋白凝胶性质的影响

研究了发芽大豆蛋白质凝胶性质的变化。采用碱提酸沉法制备大豆分离蛋白(SPI),以葡萄糖酸-δ-内酯(GDL)为凝固剂制备大豆蛋白凝胶,系统研究了不同芽长大豆蛋白凝胶强度的变化。通过SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)图谱分析了发芽过程中SPI的变化及其对大豆凝胶强度的影响。结果发现:SPI中7S球蛋白的α'、α亚基和11S球蛋白的酸性亚基A3、A发芽时发生明显降解,但11S球蛋白各亚基在发芽初期变化小,利于大豆蛋白质分子之间形成网络结构使凝胶强度增强。随着发芽时间的延长,11S球蛋白也部分发生降解,凝胶强度下降。     

Textural properties and morphology of soy 7S globulin-corn starch (amylose,amylopectin)

The texture properties (hardness, adhesiveness, cohesion, elasticity) of composite gels with different ratios (0:100, 6:94, 8:92, 10:90, 12:88, and 14:86, respectively) of soy 7S globulin to corn starch (amylose, amylopectin) were studied. Furthermore, the morphology and crystal structure of the mixed gels were investigated. The results showed: the addition of soy 7S globulin could weaken the hardness of corn starch (amylose, amylopectin) and increased the adhesiveness of corn amylose significantly (p < 0.05). Generally, when the additive amount of soy 7S globulin was 10%, the composite gels of soy 7S globulin and corn starch (amylose, amylopectin) appeared to have the best consistency, cohesion, and elasticity.     

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。     

分子间作用力对大豆蛋白凝胶形成的影响

本实验用大豆分离蛋白模拟传统豆腐的凝胶形成过程,研究分子间作用力对大豆蛋白凝胶形成过程及质构特性的影响。结果表明,大豆蛋白凝胶的形成过程受多种分子间作用力的影响,其中静电作用、疏水相互作用和氢键对石膏凝固大豆蛋白凝胶的形成具有重要的影响。     

Differential scanning calorimetry of lupin and soy proteins

Differential scanning calorimetry (DSC) was used to study the 7S and 11S globulin fractions extracted from lupin seed (Lupinus luteus) flour. In agreement with previous work on other lupin species, the isolate showed three denaturation peaks compared to the two observed with soy. By comparison with the isolated globulin fractions, the denaturation peaks at the two higher temperatures in the lupin isolate were assigned to the 11S and 7S globulins. The denaturation temperature of the lupin 7S globulin was about 10 K higher than that for the corresponding soy globulin, whereas the values for the 11S globulin were similar. All globulins displayed increasing thermal stability with decreasing moisture contents. Possible reasons for the differences in behaviour of soy and lupin protein isolates are discussed.     

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。     

Interaction of Myofibrillar and Preheated Soy Proteins

ABSTRACT: Soy protein isolate (SPI) was preheated at 90 °C and 95 °C for 3 min to obtain preheated samples, SPI₉₀ and SPI₉₅, respectively. The preheat treatment increased protein hydrophobicity and decreased the aggregation of 11S acidic and basic subunits. The 7S and 11S soy proteins exhibited a decreased thermal stability when mixed with pork myofibrillar protein isolate (MPI). The presence of preheated SPI accelerated the disappearance of myosin heavy chain in the gelling process. Incorporation of preheated SPI significantly increased the MPI gel elasticity and hardness while native SPI showed negative effects.     

Structural Characterization of Amaranth Protein Gels

ABSTRACT: Gelation capacity of a native amaranth protein isolate was studied. Structural properties of gels prepared at different protein concentration and heating conditions were analyzed. Proteins present in amaranth isolates obtained by water extraction at pH 9.0 and subsequent isoelectric precipitation are able to form gels of yellowish appearance. Gel color intensity increased while luminosity decreased with increasing protein concentrations. High protein concentration allowed the formation of matrices with high water-holding capacity. In addition, increasing the heating temperature resulted in gels of high luminosity and low water-holding capacity. The increase of protein concentration (10% to 20% w/v) as well as the increase of heating temperature (70°C to 95°C) and heating time (10 to 30 min) resulted in the formation of a more ordered matrix with smaller pores, mainly stabilized by disulfide bonds and, at a lower extent, by noncovalent interactions (specially hydrogen bonds and hydrophobic interactions). Both amaranth globulin (11S globulin and P globulin) participated in gel structure via high-molecular-weight aggregates (>100 kD). Gel structure was stabilized via noncovalent bonds by monomer species of 42 kD and those of molecular mass lower than 20 kD localized in the interstitial spaces of gel matrix.     

大豆分离蛋白亚基及7S/11S比例对肉肠品质的影响

通过采用不连续的十二烷基磺酸钠?聚丙烯酰胺凝胶电泳技术,对所选不同大豆品种中蛋白质的亚基组成及其比值(7S/11S)进行了测定,得出品种间差异对蛋白质亚基组成的变化影响较明显。然后将所选大豆品种添加到肉肠中,通过测定肉肠品质(质构、凝胶强度、得率),将其与亚基组成及7S/11S分别进行相关分析,得出各亚基对肉肠品质参数的影响显著程度各不相同。其中α-亚基和A3酸性亚基与肉肠的硬度、弹性、咀嚼性、凝胶强度均呈现显著负相关性;β-亚基与肉肠的硬度、咀嚼性、凝胶强度均呈现显著正相关性,与弹性呈现极显著的正相关性;7S/11S与肉肠的弹性呈现极显著的正相关性,与咀嚼性和凝胶强度均呈现显著的正相关性,但与硬度的相关性不显著,与得率呈现显著的负相关性。     

Investigations of Soybean 11S Protein and Myosin Interaction by Solubility, Turbidity and Titration Studies

Solubility tests, turbidity tests, and titration experiments were employed to study the possible protein-protein interactions between purified soybean 11S protein and skeletal muscle myosin and the involvement of protein subunits in the interactions. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used as an analytical tool for identification of the protein species. These tests indicate that these proteins interacted at temperatures between 85°C and 100°C. Solubility and titration experiments showed that acidic subunits of soybean US protein had little or no interaction with myosin heavy chain subunits. In contrast, soybean 11S basic subunits interacted with myosin heavy chains. The SDS-PAGE method indicated that eight commercial soy protein isolates had a similar protein species composition, but certain proteins in some isolates had lost their availability for water extraction. This may account for different functional properties exhibited by differen soy protein isolates.     

Changes in the protein complex of wheat dough affected by soybean 11 S globulin: Part 2—The interactions of soybean 11 S globulin with gluten proteins

The interactions between soy 11 S globulin and protein fractions of wheaten doughs were examined using a multistep extraction method followed by molecular sieving and characterisation of the fractions obtained. The soy 11 S globulin preparation was coupled with fluoresceine isothiocyanate to permit tracing of this protein in the complexes formed. In model experiments the average molecular weights of globulin/prolamine complexes were determined, the dispersion of light on the protein molecules being measured. The formation of high molecular weight complexes of soy 11 S globulin and prolamine was found. A shift between albumin/globulin and gluten fractions was also observed, which resulted in an increase in the contents of low molecular weight fractions dispersible in pyrophosphate buffer and acetic acid.Most of the investigations concerning the enrichment of bread with soybean proteins deal with technological problems. The documentation of the chemically induced changes in the dough-protein complex is rather limited and a number of questions are still awaiting answers. The data reported by Jakubczyk et al. (1973), Matthews (1972), Pollock & Geeds (1960a, b) do not explain the character of the interactions between soy proteins and gluten systems and give only inadequate information on the changes of molecular structure or aggregation/disaggregation phenomena occurring in the proteins of enriched dough. The results presented in our previous study (Lampart-Szczapa & Jankiewicz, 1982, encouraged us to continue the experiments to explain the rôle played by soybean 11 S globulin treated as a model soy protein in modifying the gluten matrix of wheat dough. In this paper the effects of soybean 11 S globulin on the fractional distribution of the dough-protein complex and average molecular weights of the fractions are presented.     

大豆11S球蛋白在空气-水界面上的吸附动力学

采用轴对称滴形分析法研究了大豆11S球蛋白在空气-水界面上的吸附动力学，主要检测了不同浓度和pH值条件下表面压力随吸附时间的变化。实验表明，大豆11S球蛋白在空气-水界面上的吸附随初始体相蛋白质质量分数的增加而加快，受pH值的影响尤其明显。当pH=3．0时，1S1球蛋白快速吸附到空气-水界面上；而当pH=5．0时，吸附明显下降。在吸附的初始阶段，扩散控制吸附动力学；而当表面压力较高时，蛋白质分子在界面上的展开和重排控制吸附动力学。     

Viscoelastic properties and microstructures of 11S globulin and soybean protein isolate gels: Magnesium chloride-induced gels

Heat-induced gels of 11S globulin (11S) or soybean protein isolate (SPI) were prepared using magnesium chloride (MgCl₂) as a coagulant. Viscoelastic properties and microstructures of 11S and SPI gels were quantified using dynamic viscoelastic measurement (DVM) and confocal laser scanning microscopy (CLSM). The addition of sodium chloride was necessary for 11S and SPI to form MgCl₂-induced gels. DVM indicated that 11S formed stiffer and more solid gels than SPI under all experimental conditions. CLSM showed that the microstructures of 11S gels were coarser and more heterogeneous than SPI gels in comparable conditions. The microstructures of 11S gels were highly affected by MgCl₂ concentration whereas those of SPI gels were relatively insensitive to MgCl₂ concentration. The microstructures of 11S and SPI gels were analyzed by two parameters: the fractal dimension and the average density of gel networks. Compared to SPI, 11S forms MgCl₂-induced gels with a lower fractal dimension and a higher density of network structures.     

Viscoelastic Behavior of Commercially Processed Soy Isolate Pastes During Heating and Cooling

The viscoelastic properties of soy isolate dispersions of Prolina and Brim cultivars, commercially processed, were compared during heating and cooling utilizing small deformation rheology. All isolates formed gels upon hydration. Heating to 90 °C yielded less rigid gels, as evidenced by a decreasing G'. Holding at 90 °C induced an increase in G' for Brim isolate, possibly as a result of increased hydrophobic and/or covalent bonding. Cooling to 25 °C generated a G' increase above initial levels for both cultivars, likely due to enhanced intermolecular hydrogen bonding. Mechanical spectra at 25 °C before and after heating, and at 90 °C before and after holding, confirmed these observations. A lower G' was consistently exhibited by Prolina gels throughout testing.     

Biopolymers Produced by Cross-linking Soybean 11S Globulin with Whey Proteins using Transglutaminase

Heterogeneity of biopolymers was determined by cross-linking acetylated-11S acidic subunits (Ac-11S) of soy protein with α-lactalbumin and β-lactoglobulin. The extent of polymerization was determined by electrophoresis and HPLC. Differential scanning calorimetry (DSC) was used to determine thermal properties of starting proteins and biopolymers. HPLC data demonstrated the absence of biopolymers from Ac-11S, acetylated α-lactalbumin and acetylated β-lactoglobulin when each was incubated separately with transglutaminase (TG). However, Ac-11S formed biopolymers with α-lactalbumin and β-lactoglobulin when TG was added. TG catalyzed the formation of heterologous and homologous biopolymers from whey protein isolate (WPI) and soybean 11S globulin (11S). Cross-linking WPI and 11S provided biopolymers with improved heat stability which may be useful to provide functionality to food products.     

大豆球蛋白的纤维聚集体行为及其稳定性研究

为了明确蛋白质的纤维聚集行为,本研究以大豆球蛋白(soy globulin,11S)为原料,从亚基层面对酸性条件下热诱导的11S纤维聚集过程进行跟踪,监测蛋白及其亚基的水解过程、结构变化及其稳定性。结果表明,11S的纤维化是一个多步骤的过程,包括多肽链的水解、自组装成淀粉样纤维聚集结构及逐渐生长成宏观可见的具有扭曲螺旋结构的纤维聚集体。与11S纤维化过程的单指数增长相比,酸性亚基的纤维化过程存在迟滞期。酸性亚基在纤维化聚集的初期主要贡献于纤维聚集的成核过程,碱性亚基的加入改变其纤维聚集进程。蛋白质的纤维化过程会增加11S在等电点处的溶解度,降低中性和酸性pH下的溶解度。此外,碱性环境(pH值10.0)会导致11S纤维聚集体全部溶解、宏观纤维长度变小、结构发生改变。以上研究结果旨在为合理利用蛋白纤维化聚集体作为新的功能性食品配料提供理论依据。