花生蛋白富集组分与多糖接枝对其结构特性的影响

本研究以花生脱脂粉为原料,采用低温冷沉法分离制备了花生蛋白富集组分,花生蛋白富集组分与麦芽糊精以1:1混合,在60℃湿度79%的干热条件下,花生球蛋白富集组分反应1~7 d,伴花生球蛋白富集组分反应8~24 h,运用SDS-PAGE电泳表征了花生蛋白富集组分与多糖接枝反应进行的情况,采用热特性与荧光光谱等手段表征了花生分离蛋白与多糖的接枝反应产物结构特性的变化。SDS-PAGE分析表明花生球/伴球蛋白富集组分已经在干热条件下与麦芽糊精发生交联反应生成大分子的接枝产物,伴花生球蛋白相比花生球蛋白富集组分更易与麦芽糊精发生美拉德反应。花生蛋白富集组分与麦芽糊精的混合或者接枝反应显著地改善了蛋白组分的热稳定性。花生蛋白富集组分随着糖接枝反应程度的加深其空间结构先变得松散,而后又再次变得更加紧凑。     

冷沉法制备花生球蛋白及伴花生球蛋白工艺研究

以花生球蛋白提取率为指标,对冷沉法提取花生球蛋白及伴花生球蛋白进行工艺优化。在单因素的基础上,经2次提取,通过L9(43)正交试验设计确定最佳提取工艺,结果表明,影响花生球蛋白提取率的各因素显著程度:冷沉时间料液比磷酸缓冲溶液浓度溶液pH。最佳提取条件为:料液比5∶10、溶液pH7.1、磷酸缓冲溶液浓度0.4 mol/L、冷沉时间4 h。在此条件下花生球蛋白的提取率为53.60%。花生球蛋白提取后的上清,即为伴花生球蛋白。研究发现,不同冷沉次数对花生球蛋白组分纯度影响不显著,直接干燥上清比上清酸沉后制备的伴花生球蛋白蛋白含量要低,但是组分纯度无显著差异。真空冷冻干燥制备的花生球蛋白组分纯度76.40%,伴花生球蛋白组分纯度64.10%;喷雾干燥制备的花生球蛋白组分纯度74.30%,伴花生球蛋白组分纯度62.73%,不同干燥方式对花生蛋白组分纯度影响显著(P0.01)。     

花生球蛋白和伴球蛋白的功能特性及构象研究

本文对花生球蛋白与伴球蛋白的结构和功能特性进行了分析和比较。结果表明,花生球蛋白在等电点附近(pH4.5～6.0)比伴球蛋白具有更高的溶解性,而在偏离等电点时其溶解性低于伴球蛋白,伴球蛋白的乳化活性指数(70～180 m2/g)和起泡能力(36～57%)显著高于花生球蛋白的乳化活性指数(60～130 m2/g)和起泡能力(19～33%)(P<0.05),伴球蛋白所形成热凝胶的弹性模量值(G’)约为花生球蛋白的5倍。花生球蛋白的变性温度Td(104.84℃)及焓变值ΔH(13.78 J/g)显著高于伴球蛋白的变性温度(89.47℃)与焓变值(8.11J/g)(P<0.05);花生球蛋白分子表面的巯基(SH)较少,大部分巯基包裹于球蛋白分子内部,而伴球蛋白中大部分巯基暴露于分子的表面。荧光光谱分析表明,伴球蛋白比花生球蛋白具有更疏松的三级结构和更高的界面活性。     

响应面法优化高溶解性花生蛋白提取工艺

为开发脱脂花生粕中的蛋白质,研究提取温度、提取pH值、液料比对两种花生蛋白组分——花生球蛋白和伴花生球蛋白溶解性的影响,并以响应面法设计优化提取条件,结果表明:提取温度62.93℃、提取pH9.04、液料比15.11:1(mL/g)时,两种花生蛋白溶解性最好。验证实验得到花生球蛋白的NSI为54.82%,伴花生球蛋白的NSI为74.1%,与模型预测值接近。采用响应面法对花生蛋白组分提取条件进行优化合理可行。     

高静压处理对花生蛋白的改性研究

研究高静压(300 MPa～600 MPa)处理对花生分离蛋白、花生球蛋白的溶解性、乳化性(EAI)、乳化稳定性(ESI)、表面疏水性、巯基含量等性质的影响,并对各功能性质的相互关联性进行了分析。结果表明:高静压处理可有效的改善花生分离蛋白和花生球蛋白的溶解性和乳化性,但是会降低其乳化稳定性。在400 MPa,花生分离蛋白具有最大的乳化性和表面疏水性,花生球蛋白具有最大的溶解性。在500 MPa,花生球蛋白具有最大的乳化性和表面疏水性,花生分离蛋白具有最大的溶解性;随着压力的升高,花生球蛋白和花生分离蛋白的游离巯基含量呈下降趋势。表面疏水性与乳化性存在一定的正相关。结果表明,高静压处理可以有效改善花生蛋白的功能性质。     

花生球蛋白鼠源多克隆抗体的制备及免疫学特性鉴定

为建立花生球蛋白致敏原的免疫学快速检测方法,采用碱溶酸沉法提取花生球蛋白,经SDS-PAGE检测蛋白纯度后,免疫5只6～8周龄的Balb/c雌性小鼠制备花生球蛋白鼠源多克隆抗体(多抗)血清,并进行免疫学特性鉴定。结果表明：免疫结束后得到的5个多抗血清效价均在1∶51 200以上,均具有一定的敏感性,其中2号小鼠的多抗血清敏感性较好,半数抑制浓度为320 ng/mL;制备的多抗血清特异性较强,2号小鼠的多抗血清与伴花生球蛋白、大豆球蛋白、β-伴大豆球蛋白、牛血清白蛋白、麦朊蛋白、乳清蛋白的交叉反应率均较低,在0.5%以下;Western blot鉴定结果表明成功制备出了花生球蛋白鼠源多克隆抗体。综上,获得了效价高、敏感性强、特异性优良的花生球蛋白鼠源多克隆抗体,为其单克隆抗体的制备及免疫学快速检测方法研究奠定了基础。     

花生蛋白组分及其功能性质研究进展

介绍了花生蛋白的分类、氨基酸及亚基组成;同时对花生球蛋白、伴花生球蛋白Ⅱ和伴花生球蛋白Ⅰ等主要组分的提取方法--冷沉淀法和硫酸铵沉淀法进行总结;此外对花生蛋白及其主要组分的溶解性、乳化性、起泡性、凝胶性等功能性质进行综述。并归纳花生蛋白研究存在的问题,提出今后的发展方向。     

多糖接枝对花生球蛋白功能特性的影响

本文研究了花生球蛋白与葡聚糖能够在一定控温控湿的条件下进行干热反应,生成美拉德反应的多糖接枝产物。通过荧光光谱、圆二色谱、差示扫描量热法和紫外分光光度法等先进的检测分析手段对美拉德反应对接枝产物结构特性的影响进行表征,同时也阐述了接枝产物的结构变化与其功能特性改变之间的构效机理。研究结果表明花生球蛋白酸性亚基比碱性亚基更易与葡聚糖发生接枝反应。对花生球蛋白的预加热处理不能提高其与葡聚糖的反应速度。花生球蛋白与热处理花生球蛋白在与多糖接枝反应过程中,其三级结构皆变得更加紧凑,限制了蛋白与多糖的接枝反应速度与反应程度。未经热预处理的花生球蛋白与多糖的接枝产物具有很高的溶解性和乳化活性,在干热反应第14 d其溶解性和乳化活性指数达到实验条件下的最高值,分别为95%和149 m2/g。     

大豆β-伴球蛋白的功能特性分析

从6种优质大豆中提取了大豆β-伴球蛋白,SDS-PAGE电泳后均显示出6条谱带,并对它们的功能特性进行研究。研究表明：大豆β-伴球蛋白持水性为市售大豆分离蛋白的1.15～1.48倍,乳化性和乳化稳定性分别为市售大豆分离蛋白粉的1.04～2.35倍和1.21～4.27倍,溶解性为市售大豆分离蛋白粉的1.33～2.57倍,大豆β-伴球蛋白具有较好的功能特性。     

不同品种花生蛋白主要组分及其亚基相对含量分析

对我国主要花生种植区的170个花生品种的蛋白质组成进行SDS-PAGE电泳分析。结果表明:不同花生品种的蛋白组分和亚基相对含量在种质材料间存在差异。各品种均含有花生球蛋白和伴花生球蛋白,花生球蛋白与伴花生球蛋白比值变化范围介于0.80～1.68之间,变异系数为15.23%,大于10%,说明不同花生品种蛋白组成比例存在较大的变异性。各品种亚基变异显著,其中,花生球蛋白的35.5kD亚基差异最显著,变异系数达55.07%,双纪2号等35个品种缺失35.5kD亚基条带,占所测试品种的20.59%。相关性分析表明,花生球蛋白与伴花生球蛋白之间呈极显著负相关(r=-0.998)。     

Effects of transglutaminase catalyzed crosslinking on physicochemical characteristics of arachin and conarachin-rich peanut protein fractions

Arachin and conarachin-rich fractions of peanut protein were extracted by using cryoprecipitation followed by centrifugation. These two fractions were individually crosslinked using transglutaminase (TG). The physicochemical characteristics including aggregation due to crosslinking, solubility, thermal denaturation temperature (T_d) and denaturation enthalpy (△ H), morphology of microstructure and surface hydrophobicity (H₀) of TG-treated and untreated arachin and conarachin-rich fractions were determined. The relative contents of arachin and conarachin in arachin and conarachin-rich fractions were 75% and 65%, respectively. Conarachin-rich fractions were found to more conveniently aggregate than arachin-rich from the results of SDS-PAGE. The solubility of treated arachin and conarachin-rich fractions was decreased by 66.13% and 36.91%, respectively. Only marginal increase in T_d was observed in the case of crosslinked arachin-rich fraction while significant increase in T_d (of the order of 10 °C) was observed in crosslinked conarachin-rich fraction. The H₀ values of both crosslinked arachin and conarachin-rich fractions decreased significantly. The treated arachin and conarachin-rich fractions had more compact microstructure compared to their untreated samples. Hence, the comparison of arachin and conarachin-rich in crosslinking and the mechanism of properties enhancement by TG is the aim of the research.     

Enzymatic hydrolysis and their effects on conformational and functional properties of peanut protein isolate

The effects of limited enzymatic hydrolysis by Alcalase on the conformational and functional properties of peanut (Arachis hypogaea L.) protein isolate (PPI) were investigated. Acid subunits of arachin were most susceptible to Alcalase hydrolysis, followed by conarachin and the basic subunits of arachin. Enzymatic hydrolysis increased the thermal stability of arachin and led to a sharp increase in the number of disulphide bonds with a decrease of the sulphydryl group in PPI hydrolysates in comparison with PPI. The analysis of intrinsic fluorescence spectra indicated a more moveable tertiary conformation of PPI hydrolysates than PPI. The limited emzymatic hydrolysis improved the functional properties of PPI, such as protein solubility and gel-forming ability, but impaired the emulsifying activity index.     

Improvement of functional properties of peanut protein isolate by conjugation with dextran through Maillard reaction

Reaction mixtures containing peanut protein isolate (PPI) and dextran (1:1 weight ratio) were dry-heated at 60 °C and 79% relative humidity for 7 days. SDS-PAGE analysis indicated that PPI had become complexed with dextran to form conjugates of higher molecular weight. Arachin, accounting for 50% of peanut proteins, was hard to glycosylate with dextran, which might limit the extent of glycosylation of PPI. The thermal stability of PPI was remarkably improved by mixture/conjugation with dextran. Proteins in mixtures/conjugates might have a more compacted tertiary conformation than PPI. The protein solubility of conjugates at pH 4.5-6.0 was remarkably increased compared with the PPI/mixture. Mixture with dextran could significantly improve the emulsifying and foaming properties of PPI (p < 0.05). Conjugation with dextran could further enhance emulsifying and foaming properties of PPI.     

花生湿热处理对其分离蛋白的结构和功能特性的影响

本文通过还原/非还原电泳、热差示扫描、表面疏水性及等电点测定等手段,表征了花生湿热处理对其分离蛋白结构和功能特性的影响.结果表明:在105℃随着湿热处理时间延长其分离蛋白中伴球蛋白所占比例逐渐减少,球蛋白在加热75min后产生热聚集,此外通过美拉德反应生成的糖蛋白含量逐渐增加,分离蛋白等电点向酸性偏移,表面疏水性先逐渐...     

Functional properties of protein fractions obtained from commercial yellow field pea (Pisum sativum L.) seed protein isolate

Commercial pea protein isolate was separated into water-soluble (WS), salt-soluble (SS), alkaline-soluble (AS) and ethanol-soluble (ES) fractions. AS fraction was the most abundant, constituting about 87% of the proteins in PPI followed by WS, SS and ES fractions in decreasing order. ES fraction consistently formed emulsions with a narrow range of smaller oil droplet sizes (0.6–19 μm) at pH 4.0, 7.0 or 9.0 compared to a wider range of sizes for emulsions stabilised by WS, SS and AS fractions. Emulsions formed with ES fraction were also the most stable (p < 0.05) over the 3 h test period at all the pH values used in this work. The WS fraction had significantly highest (p < 0.05) protein solubility and foaming capacity at all the pH values when compared to solubility of PPI, SS, and ES. Except for AS and ES fractions, foaming capacities of the protein fractions were higher at pH 9.0 than at pH 4.0 or 7.0.     

Effect of denaturation during extraction on the conformational and functional properties of peanut protein isolate

Peanut protein isolate (PPI) was extracted by alkali dissolution and acid precipitation from defatted peanut flour. The effects of extraction conditions on the denaturation and functional properties of PPI were investigated. In comparison with native peanut protein (NPP) which was extracted by ammonium sulfate, the PPI extracted by alkali dissolution and acid precipitation had a higher extent of denaturation. Arachin was affected more easily by the extraction process than conarachin and led to a noticeable decrease of thermal stability of PPI. PPI contained much lower sulfhydryl and disulfide bond contents than NPP. The analyses of intrinsic fluorescence spectra indicated a more compacted tertiary conformation of NPP than PPI. Extraction process influenced the functional properties of PPI, such as protein solubility, emulsifying activity index and foaming capacity. The relatively poor functional properties of PPI might be associated with protein denaturation/unfolding and subsequent protein aggregation.

Industrial relevance

Peanut is an important oilseed crop and a well-accepted food. After oil production through thermal treatment, the defatted peanut flour is the main byproduct, which possesses a large amount of proteins. However, due to the low extraction yield and poor functional properties of these proteins, they are not well utilised in industry till now. In this work, peanut proteins were extracted by two techniques. The results indicated that extraction technique could significantly modify the functional properties of peanut proteins. Therefore, this work is helpful for industrial utilisation of peanut proteins.     

Surface functional properties of blood plasma protein fractions

The solubility, foaming and emulsifying properties of porcine blood plasma and its main protein fractions (serum, globulins and albumin) were investigated at pH 4.5, 6.0 and 7.5 in order to clarify the contribution of each fraction and encourage the optimisation of plasma-derived products. Soluble protein contents above 85% were obtained in all samples. Plasma, serum and albumin showed good foaming capacities, reasonably similar at different pH conditions, although the highest foam stability corresponded to both albumin and plasma at pH 4.5 and 6.0. All protein fractions showed good emulsifying activities, but the stability of the formed emulsions decreased with acidification, being emulsions of albumin and globulins at pH 7.5 the most stable ones. In addition, the interaction indexes calculated to investigate protein–protein interactions revealed synergistic interactions between albumin and globulins when in co-occurrence in their foaming capacity at pH 6.0 and 7.5, and in the stability of emulsions at pH 4.5 and 6.0, but slightly negative effects in the solubility of the mixture, and a great decrease in the stability of emulsions at pH 7.5. On the other hand, the elimination of fibrinogen improved the stability of emulsions and foams at acidic conditions.     

红衣组分对花生分离蛋白及其酶解产物理化和抗氧化性质的影响

以花生为原料,研究了花生红衣组分对花生分离蛋白及其酶解产物理化和抗氧化性质的影响。研究结果表明:含红衣的分离蛋白酶解速度低于不含红衣的;相同水解度条件下,含红衣的表面疏水性小于脱红衣的;GPC分布中,含红衣的峰值大于脱红衣的;红衣组分能够提高花生分离蛋白及其酶解产物的溶解性、乳化稳定性、乳化活性;在相同水解度条件下,含红衣的水解产物多酚含量显著高于脱红衣的,多酚含量的差异与花生分离蛋白及其水解产物的抗氧化性具有正相关性。     

响应面优化转谷氨酰胺酶改性花生分离蛋白工艺的研究

为了改善花生分离蛋白的凝胶特性,研究了利用转谷氨酰胺酶交联改性花生分离蛋白的工艺。在进行了酶添加量、花生分离蛋白浓度和酶作用时间单因素试验基础上,利用响应面试验设计优化了酶交联改性的最佳条件。并分别测定了酶改性前后花生分离蛋白的功能性,包括:溶解性、吸油性、持水性、乳化性和乳化稳定性、起泡性和起泡稳定性。通过响应面分析得到酶改性的最佳条件:酶添加量、花生分离蛋白质量浓度和酶作用时间分别为17.75 U/g、29.60 g/mL和376 min,在此条件下,凝胶的硬度可达到333.49 g。经转谷氨酰胺酶改性后,花生分离蛋白的吸油性和持水性均有不同程度的提高,分别提高了27.41%和61.24%。