Coagulation of soy protein isolates induced by subtilisin Carlsberg

The coagulation of soy protein isolates (SPI) induced by subtilisin Carlsberg was studied. The proteins were digested to fragments of 16 kDa or less in the early stage of the reaction, followed by coagulation. The time-course of the coagulation measured by turbidity was separated into three phases. The turbidity decreased from the initial level observed at time zero to the minimum level (OD1) at time T1 (15-20 min) in the first phase. Then, it increased drastically to reach the maximum (OD2) at time T2 (60-70 min) in the second phase, which was followed by a slight decrease in the third phase. The coagulation was terminated at T2, where 30-35% of the weight of the SPI proteins was in coagula. Proteins in the coagula were degraded slowly in the prolonged incubation, and the protein content in the coagula was finally 15-20% of the weight. The time-course of the turbidity agreed well with that of the weight of the precipitates formed, indicating that the turbidity reflects the progress of the coagulation. The turbidity change (OD1 to OD2) from the start to the end of the coagulation increased proportionally to the SPI concentration (4.9-11 mg/mL), although the time (T1 to T2) needed for the coagulation was independent of the concentration. The growth of the coagula is promoted by increasing the SPI concentration and is rate-limiting in the coagulation.     

玄武岩纤维改性大豆蛋白胶黏剂性能及胶合机理

为提高大豆蛋白胶黏剂的胶合强度,开发环境友好型木材用生物质胶黏剂,采用玄武岩纤维(basalt fiber,BF)作为增强改性剂,成功制备了胶接性能优良的玄武岩纤维改性大豆蛋白胶黏剂(basalt fiber/soybean protein isolate adhesive,BF/SPIA),并对BF/SPI的胶接强度、流变特性、化学基团及形态结构进行了分析。研究结果表明:随着玄武岩纤维用量的逐步提高,改性大豆蛋白胶黏剂的力学强度呈先增加后下降的趋势。当BF质量分数为5%时,BF/SPIA干、湿状胶接强度分别达2.15和0.92 MPa。流变特性测试结果表明BF/SPI胶黏剂的剪切模量有所增加;傅里叶红外光谱(fourier infrared spectrum,FTIR)显示出改性大豆蛋白胶黏剂中亲水基团降低,Si—O键特征峰明显,BF与大豆蛋白分子间相互作用,形成新的复合结构;扫描电镜(scanning electron microscope,SEM)图谱印证了BF与SPAI两相体系融合性良好,且固化胶接界面较为致密。中试试验结果进一步表明玄武岩纤维改性大豆蛋白胶黏剂具有良好的稳定性,其湿状胶接强度可高达1.05 MPa,木材压缩率仅14.92%,实用性较强。该研究可为改性大豆蛋白胶黏剂的工业化应用及推动环保木材胶黏剂的应用进程提供理论参考。     

高水分大豆蛋白组织化生产工艺和机理分析

以低温豆粕为原料，应用带冷却工艺的双螺杆挤压实验室工作站，开发出高水分组织化大豆蛋白产品的生产工艺；分析了螺杆转速、物料湿度、喂料速度和机筒温度等操作参数对产品质构和色泽的影响规律；利用扫描电镜对高水分组织化大豆蛋白产品的微观结构进行了观察和分析。结果表明：机筒温度在150℃左右时，产品具有较好的组织化度、色泽及口感；随着物料水分的增加，产品的组织化度、粘着性和明度指数逐渐增加，而硬度和咀嚼度逐渐降低；随着螺杆转速的增加，产品的组织化度、硬度、粘着性和咀嚼度均有增加的趋势；喂料速度对产品的表观形态影响较大，喂料速度较大时，产品表观粗糙、组织化度降低。单因素实验优选的操作参数为：机筒温度145℃～155℃，物料湿度45％～50％，螺杆转速90～160 r/min，喂料速度20～40 g/min。结合对生产工艺和产品微观结构分析，提出了高水分组织化大豆蛋白产品形成过程的“膜状气腔”理论假设，以供讨论和指导生产实践。     

Incorporation of soy milk lipid into protein coagulum by addition of calcium chloride.

Lipid holding in tofu (soybean curd) has such stability that it is not released even when it is cooked. The lipid in soy milk is incorporated into the tofu with coagulation of proteins on processing by the addition of a calcium salt. In this study, the incorporation of lipid into the coagulum was examined from the association with protein. The lipid in soy milk was obtained as the floating fraction by centrifugation. The floating fraction decreased by the addition of calcium chloride prior to forming a protein coagulum. When half of the protein coagulated, all of the floating fraction became inseparable. The protein in soy milk, then, was separated into particulate and soluble fractions by centrifugation. The decrease of the floating fraction with added calcium chloride was parallel to the coagulation of the particulate protein. The association of the floating fraction and the soluble protein occurred after the new particles formed from the soluble proteins. These results indicated that the lipid incorporation took place by the conjugation of the lipid and protein particles.     

Fast characterization of industrial soy protein isolates by direct analysis with matrix-assisted laser desorption ionization time-of-flight mass spectrometry

Industrial soy protein isolates (SPIs) due to differences in their processing conditions may differ both in composition and in degree of hydrolysis. As a result, they display different performance in food production and final food properties like consistency and taste. To address this issue, a fast, cheap, and simple method for screening and characterization is required. In this article, the successful analysis of soy protein isolates, a complex mixture of proteins with glycinin and beta-conglycinin as major components, by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) is demonstrated. The preparation implements a fast extraction of the proteins from the raw SPI either under neutral or reducing conditions. The extracts are analyzed subsequently by MALDI-TOF-MS without further purification. Results of the two conditions are compared. Finally, different SPIs from different suppliers are analyzed and compared concerning their consistency. The method could be applied to other plant proteins and mixtures thereof. Since the composition and intactness of different subunits play important roles in functional properties of soy proteins, rapid methods for fingerprinting of different industrial soy protein sources will be valuable tools for successful product formulation.     

基于模型化合物的糠醇改性大豆蛋白基胶黏剂机理研究

为了开发真正意义上的环保型大豆蛋白基胶黏剂,该文以糠醇作为交联剂,以组成蛋白质分子的二肽为起点,采用模型化合物思想研究大豆蛋白基胶黏剂的交联改性基础理论,借助核磁共振C谱和电喷雾电离质子仪分析糠醇交联改性大豆蛋白基胶黏剂的机理。同时通过检测大豆蛋白胶胶合板性能,分析大豆蛋白基胶黏剂制备基础理论与实际性能之间的关系。研究结果表明:1)糠醇与丙谷二肽的共缩聚反应和糠醇的自缩聚反应,两者为竞争关系;2)一般碱性条件下,糠醇极其稳定,两者都不明显,只有在强碱性条件下糠醇可能产生活性稍大的酰胺负离子与丙谷二肽发生一定的共缩聚,反应速率较慢,实现共缩聚的时间太长,但自缩聚反应依然不明显;3)糠醇与丙谷二肽的共缩聚反应和糠醇的自缩聚反应都需要较强的酸性,且前者低于后者,酸性太强时糠醇自缩聚占主导,酸性太弱时产生的糠醇碳正离子浓度和反应活性较弱,不利于自缩聚和共缩聚;在pH值为3时,共缩聚表现出较强的竞争性。4)糠醇与丙谷二肽共缩聚反应和糠醇自缩聚反应主要发生在酸性条件下,糠醇与丙谷二肽发生共缩聚反应位点首先是端酰胺基团,其次是伯胺,而肽键氨基由于存在较大的空间位阻导致几乎不参与反应。5)糠醇与大豆蛋白模型化合物反应机理与大豆蛋白基胶黏剂实际性能具有较强的对应性,即在pH值为3时,糠醇与大豆蛋白基胶黏剂反应程度较高,表现为较强的耐水性和热稳定性。该研究工作的开展对实现大豆蛋白基胶黏剂的科学使用,提升大豆蛋白基胶黏剂的市场竞争力具有重要意义。     

Effects of ultrasound pretreatment on the enzymatic hydrolysis of soy protein isolates and on the emulsifying properties of hydrolysates

Soy protein isolate (SPI) was modified by ultrasound pretreatment (200 W, 400 W, 600 W) and controlled papain hydrolysis, and the emulsifying properties of SPIH (SPI hydrolysates) and USPIH (ultrasound pretreated SPIH) were investigated. Analysis of mean droplet sizes and creaming indices of emulsions formed by SPIH and USPIH showed that some USPIH had markedly improved emulsifying capability and emulsion stabilization against creaming during quiescent storage. Compared with control SPI and SPIH-0.58% degree of hydrolysis (DH), USPIH-400W-1.25% (USPIH pretreated under 400W sonication and hydrolyzed to 1.25% DH) was capable of forming a stable fine emulsion (d43=1.79 μm) at a lower concentration (3.0% w/v). A variety of physicochemical and interfacial properties of USPIH-400W products have been investigated in relation to DH and emulsifying properties. SDS-PAGE showed that ultrasound pretreatment could significantly improve the accessibility of some subunits (α-7S and A-11S) in soy proteins to papain hydrolysis, resulting in changes in DH, protein solubility (PS), surface hydrophobicity (H0), and secondary structure for USPIH-400W. Compared with control SPI and SPIH-0.58%, USPIH-400W-1.25% had a higher protein adsorption fraction (Fads) and a lower saturation surface load (Γsat), which is mainly due to its higher PS and random coil content, and may explain its markedly improved emulsifying capability. This study demonstrated that combined ultrasound pretreatment and controlled enzymatic hydrolysis could be an effective method for the functionality modification of globular proteins.     

Effect of storage temperature and water activity on the content and profile of isoflavones, antioxidant activity, and in vitro protein digestibility of soy protein isolates and defatted soy flours

The aim of this work was to study the effect of 1 year of storage at different temperatures and 1 month of storage at different water activities on the content and profile of isoflavones, antioxidant activity, and in vitro protein digestibility of defatted soy flours (DSF) and soy protein isolates (SPI). The storage for up to 1 year, at temperatures from -18 to 42 degrees C, had no effect on the total content of isoflavones, but the profile changed drastically at 42 degrees C, with a significant decrease of the percentage of malonylglucosides with a proportional increase of beta-glucosides. A similar effect was observed for SPI stored at a(w) = 0.87 for 1 month. For DSF, however, there was observed a great increase in aglycons (from 10 to 79%), probably due to the action of endogenous beta-glucosidases. The antioxidant activity decreased after storage at 42 degrees C, but the in vitro protein digestibility did not change.     

Proteomic profiling of the coagulation of milk proteins induced by chymosin

Chymosin-induced coagulation of individual milk proteins during incubation at 30 °C was investigated using a proteomic approach. The addition of chymosin (0.006 units/mL) caused the milk proteins to coagulate after a 3 h incubation period. Approximately 88% of the milk proteins were coagulated into the milk pellet fraction, and the protein concentration of the milk supernatant fraction (MSF) decreased from 29.88 ± 0.12 to 3.74 ± 0.13 mg/mL. SDS-PAGE analysis showed that α(S)-, β- and κ-caseins in the MSF were almost depleted and that the total intensity of the protein bands corresponding to α(S)-caseins (α(S1) and α(S2)), β-casein, and κ-casein decreased from 1088.0, 901.5, and 617.0 area units to 6.9, 6.1, and 5.2 area units, respectively. Two-dimensional electrophoresis analysis indicated that α(S1)-, α(S2)-, β-, and κ-casein and a fraction of the β-lactoglobulin and serum albumin were found in the MSF following incubation with chymosin.     

大豆分离蛋白成膜性研究

该文通过单因素试验和正交试验对大豆分离蛋白的成膜性进行研究。结果表明，大豆分离蛋白浓度、增塑剂(甘油)、还原剂(Na₂SO)、交联剂(谷氨酰胺转胺酶)对膜的性能有较大影响，最佳处理条件为：大豆分离蛋白5.0%、甘油1.5%、Na₂SO₃ 0.1%、TG酶0.2%，大豆分离蛋白浓度对膜性能影响最大，其次是甘油含量，Na₂SO₃和TG酶的浓度对膜性能影响较小。     

Protein digestibility-corrected amino acid scores (PDCAAS) for soy protein isolates and concentrate: criteria for evaluation

Protein quality, as determined by the PDCAAS method, is a measure of a protein's ability to provide adequate levels of essential amino acids for human needs. PDCAAS is calculated using an amino acid profile and true digestibility of a food protein. Soy protein is recognized as a high quality plant protein, but published PDCAAS values may vary based on the soy protein ingredient as well as the reproducibility and accuracy of the testing methods. Comparison of PDCAAS values for four differently processed soy ingredients, including three isolated soy proteins (ISP) and one soy protein concentrate (SPC), was made using two different laboratories with evaluation of the impact of the reproducibility and accuracy of amino acid profiles. PDCAAS calculations, using amino acid values from one laboratory, yielded a truncated PDCAAS of 1.00 for all four ingredients, while a second laboratory provided statistically significantly lower scores (0.95-1.00). We conclude that analytical method error can be a significant contributor to PDCAAS differences and can be mitigated by the application of amino acid nitrogen recovery correction factors.     

水热预处理提高花生分离蛋白酶解效率及其机理分析

为了提高花生分离蛋白的酶解效率,该文采用水热法对花生分离蛋白进行预处理,利用响应面试验设计优化预处理工艺,并研究比较了预处理前后花生分离蛋白酶解敏感性和空间构象的变化。结果表明:优化后的最佳预处理条件为水热压力90 MPa、水热温度85℃、水热时间20 min,此条件下酶解产物水解度达到16.3%,相对未经预处理的酶解产物提高了8.1个百分点。水热预处理提高了花生分离蛋白的主要组分花生球蛋白和伴球蛋白的酶解敏感性,使酶解产物蛋白质回收率提高了31.9个百分点。进行荧光光谱和红外光谱分析发现水热预处理使花生分离蛋白三级结构展开、二级结构紧密程度下降,可能是其酶解敏感性提高的主要原因。因此水热预处理是一种辅助提高花生分离蛋白酶解效率行之有效的方法。     

两级泡沫分离废水中大豆蛋白的工艺

为了有效处理高浓度大豆蛋白废水,在研究初始pH值、表观气速和温度特别是在较高温度（60℃）对泡沫分离大豆蛋白废水效果影响的基础上,建立了可同时提高富集比和回收率的两级泡沫分离工艺。在60℃时,对大豆蛋白质量浓度为4.0 g/L的废水泡沫分离,富集比比常温时提高4倍多。两级泡沫分离工艺中,第一级分离在大豆蛋白质量浓度为4.0 g/L,初始pH值为7.0,表观气速0.133 cm/s,60℃下操作,富集比为7.71,残液作为第二级泡沫分离的进料;第二级分离先在20℃,表观气速0.133 cm/s下进行,待塔顶     

Properties of cast films from hemp (Cannabis sativa L.) and soy protein isolates. A comparative study

The properties of cast films from hemp protein isolate (HPI) including moisture content (MC) and total soluble mass (TSM), tensile strength (TS) and elongation at the break (EAB), and surface hydrophobicity were investigated and compared to those from soy protein isolate (SPI). The plasticizer (glycerol) level effect on these properties and the interactive force pattern for the film network formation were also evaluated. At some specific glycerol levels, HPI films had similar MC, much less TSM and EAB, and higher TS and surface hydrophobicity (support matrix side), as compared to SPI films. The TS of HPI and SPI films as a function of plasticizer level (in the range of 0.3-0.6 g/g of protein) were well fitted with the exponential equation with coefficient factors of 0.991 and 0.969, respectively. Unexpectedly, the surface hydrophobicity of HPI films (including air and support matrix sides) increased with increasing the glycerol level (from 0.3 to 0.6 g/g of protein). The analyses of protein solubility of film in various solvents and free sulfydryl group content showed that the disulfide bonds are the prominent interactive force in the HPI film network formation, while in the SPI case, besides the disulfide bonds, hydrogen bonds and hydrophobic interactions are also to a similar extent involved. The results suggest that hemp protein isolates have good potential to be applied to prepare protein film with some superior characteristics, e.g., low solubility and high surface hydrophobicity.     

Effect of flavor compound chemical structure and environmental relative humidity on the binding of volatile flavor compounds to dehydrated soy protein isolates

Influence of flavor compound chemical structure, including functional group and stereochemistry, and environmental relative humidity (RH) on the binding of volatile flavor compounds to dehydrated soy protein isolates (SPIs) was evaluated by inverse gas chromatography. Binding of selected volatile flavor compounds differed slightly between SPIs of different origins. Results showed that the flavor compound chemical structure greatly determined its binding potential to SPIs. Binding of nonpolar flavor compounds (hydrocarbon) to soy proteins was attributed mainly to nonspecific van der Waals dispersion forces and was not affected by adsorbed water. The more polar flavor compounds (ester, ketone, aldehyde, and alcohol) exhibited both specific (hydrogen bonding, dipole forces) and nonspecific interactions, and their binding with soy proteins was greatly impaired by adsorbed water in the extremely low humidity region (approaching 0% RH). Further water uptake in the 30 to 50% RH region did not significantly affect the binding of polar compounds, although sorption of alcohol compounds (when present at high levels) further increased.     

Optimization of the enzymatic deamidation of soy protein by protein-glutaminase and its effect on the functional properties of the protein

The effects of enzymatic deamidation by protein-glutaminase (PG) on the functional properties of soy protein isolate (SPI) were studied. Conditions for the deamidation were evaluated by means of response surface methodology (RSM). Optimal conditions based on achieving a high degree of deamidation (DD) with a concurrently low degree of hydrolysis (DH) were 44 °C, enzyme:substrate ratio (E/S) of 40 U/g protein and pH 7.0. Under optimal conditions, both DD and DH increased over time. SDS-PAGE results indicated that lower molecular mass subunits were produced with increasing DD. Far-UV circular dichroism spectra revealed that the α-helix structure decreased with higher DD, while the β-sheet structure increased until 15 min of deamidation (32.9% DD), but then decreased at higher DD. The solubility of deamidated SPI was enhanced under both acidic and neutral conditions. SPI with higher DD showed better emulsifying properties and greater foaming capacity than SPI, while foaming stability was decreased. It is possible to modify and potentially improve the functional properties of SPI by enzymatic deamidation using PG.     

适宜含水率保持油茶籽贮藏品质

为了确定油茶籽贮藏适宜的含水率,研究了在4℃,不同含水率(7%、10%、13%、16%、20%)油茶籽贮藏期间的品质变化。结果表明,较低的含水率能较好保持油茶籽的贮藏特性及营养品质。其中,含水率为7%的油茶籽贮藏效果较好,但与10%处理效果差异不明显(P>0.05)。在整个贮藏期,含水率为7%时油茶籽可溶性蛋白下降了13.05 mg/g,油酸含量下降了2.38%,酸值、过氧化值等品质指标上升速率较慢,同时能较好保持β-谷甾醇和角鲨烯等生物活性成分;其次是10%的含水率处理。而含水率为20%的油茶籽贮藏期间可溶性蛋白下降较快,贮藏结束时为25.47 mg/g,油茶籽劣变严重,所提取的油样品质变差,营养物质含量较少,因此含水率20%的油茶籽不适宜长期贮藏。综合考虑油茶籽品质因素和处理成本,认为控制含水率在10%以下能较好保持油茶籽的贮藏品质。该研究可为科学合理地贮藏油茶籽提供参考。     

糖接枝处理改善大豆蛋白纤维聚集体泡沫稳定性

为了探究糖接枝对大豆蛋白纤维聚集行为和泡沫性质的影响,明确蛋白质结构与功能的关系,该研究以大豆蛋白（soy protein isolation,SPI）和乳糖（lactose）为原料,通过干热法制备糖接枝大豆蛋白（SPI-lactose conjugate,SPI-Lac）,以及在酸性条件下加热诱导其形成纤维聚集体（pH值2.0）,制备了一种糖接枝大豆蛋白纤维聚集体（SPI-lactose conjugate fibillar aggregates）,并考察了糖接枝对大豆蛋白的纤维聚集行为及泡沫性质的影响。研究结果表明：大豆蛋白在酸性条件下（pH值2.0）经加热后会发生水解,同时水解产物不断聚集形成大分子的纤维聚集体。糖接枝导致大豆蛋白的水解速度下降,但荧光光强和粒径的结果表明糖接枝能增强纤维聚集能力。SPI-Lac在中性条件下的溶解度（pH值5.0-7.0）显著高于SPI（P<0.05）,且不同时间处理的SPI-Lac纤维聚集体均能改善SPI在酸性条件下的溶解度（pH值2.0-5.0）。此外,不同时间处理的SPI-Lac纤维聚集体在酸性条件下的起泡能力均高于SPI纤维聚集体。SPI和SPI-Lac纤维聚集体的形成会导致SPI起泡能力的下降,但是短时间酸热处理形成的纤维聚集体泡沫稳定性得到显著改善。因此,糖接枝结合短时间酸热处理制备的糖接枝大豆蛋白纤维聚集体在中性条件下的泡沫稳定性显著提高（P<0.05）,是合理有效的蛋白质改性方法。