Stepwise extraction of proteins and carbohydrates from soybean seed

The stepwise hot water extraction of soybeans, which were extractions in a series of procedures of whole soybean seeds, dehulled and sliced ones, and pressed ones carried out by autoclaving, was investigated to study the localization in the seed and their characteristics. The characteristics of each extraction were studied by HPLC, SDS-PAGE, components analysis, microscopic observation, and effect for some enzymes. Carbohydrates were easier to extract than protein. In the extractions, the ratio of uronic acid per total sugar was constantly about 0.3. A comparison of these extracts, soybean milk, extraction from defatted soybean meal, and soybean milk residues was also carried out, and the characteristics and the localization were investigated. Mid-sized proteins in soybean milk were easy to extract. However, hardly any high molecular weight proteins or high molecular weight carbohydrates were extracted. The proteins and carbohydrates were considered to be localized in the middle lamella and in the protein and/or oil bodies of the cell, and the proteins and carbohydrates were gradually extracted through seed and cell breaking. Gelation was observed only in the boiled extracts from whole seeds. Pepsin and trypsin digests of the high molecular weight protein had inhibitory activity against the angiotensin I converting enzyme.     

Isolation and enzymatic digestion of body complex of soybean seed

The body complex of the soybean seed (BCSS) was isolated from the single cells (27.2%) by a sequential procedure of autoclaving with water, cellulase digestion for the primary cell wall, pectinase digestion for the secondary cell wall, and defatting with hexane washing. Its characteristics were then investigated. The defatted BCSS (DBCSS) consisted of protein (76.5%) and mannose-rich carbohydrates (3.2%). Screening of the food-processing protease for the digestion of DBCSS was carried out, and a kind of alkaline protease was selected. The inner protein of DBCSS was easily extracted with 0.1 M sodium carbonate buffer, pH 10, and the insoluble shell of the body complex (SDBCSS) was left. SDBCSS consisted of hydrophobic amino acid-rich protein. SDBCSS was easily digested by the selected alkaline protease. SDBCSS was dissolved by boiling with sodium dodecyl sulfate-mercaptoethanol, and it was found to consist of a protein of approximately 3 kDa. The high enzymatic digestion including the selected protease for soybean seed and defatted soybean meal was carried out; both were extracted and digested with a yield of >99.5%. The final indigestible residue was found as paired hexagonal and filamentous organs of the soybean cells.     

Compositional analysis of glyphosate-tolerant soybeans treated with glyphosate.

The compositional analyses and safety assessment of glyphosate-tolerant soybeans (GTS) were previously described. These analyses were extensive and included addressing the potential effects on seed composition from the genetic modification. Detailed compositional analyses established that GTS, which had not been treated with glyphosate, were comparable to the parental soybean line and to other conventional soybeans. In this study, two GTS lines, 40-3-2 and 61-67-1, were treated with commercial levels of glyphosate, the active ingredient in Roundup herbicide. The composition of the seed from soybeans sprayed with glyphosate was compared to that of a nonsprayed parental control cultivar, A5403. The nutrients measured in the seed included protein, oil, ash, fiber, carbohydrates, and amino acids. The concentration of isoflavones (also referred to as phytoestrogens) was also measured as these compounds are derived from the same biochemical pathway that was engineered for glyphosate tolerance. The analytical results from these studies demonstrate that the GTS soybeans treated with glyphosate were comparable to the parental soybean cultivar, A5403, and other conventional soybean varieties.     

Proteomic analysis of high protein soybean (Glycine max) accessions demonstrates the contribution of novel glycinin subunits

Limited biochemical information is available on soybean accessions that have seed protein content greater than 45% of the seed dry weight. SDS-PAGE analysis of seed proteins from nine soybean accessions revealed significantly higher amount of seed storage proteins in these accessions when compared with that of soybean cultivar Williams 82. High-resolution two-dimensional gel electrophoretic analysis of seed proteins revealed significant differences among several seed storage protein components in these accessions. A total of 51 protein spots were identified using peptide mass fingerprinting (MALDI-TOF MS). The contribution of these proteins to the overall protein content of the accessions was quantified using Delta2D image analysis software. Results showed that among the majority of the nine accessions, the largest difference in higher protein quantity was within the seed 11S storage globulins. The high protein trait from PI407788A was successfully transferred to an experimental line, LG99-469, demonstrating that this trait was transferable and robust.     

超声辅助提取辣椒籽蛋白工艺优化

以辣椒籽为原料,采用超声辅助法提取辣椒籽蛋白并利用响应面优化法提取工艺,在单因素试验的基础上,选取p H值,提取时间,超声功率,料液比4个因素进行响应面试验,根据所得试验结果确定最佳提取条件为:p H值11,提取时间13.31 min,超声功率336.21 W,料液比1∶35.85,在此条件下蛋白的预计提取量为5.90 g/(100 g)。利用Design expert软件对影响辣椒籽蛋白提取量的主要因素及其相互作用进行探讨,结果由大到小依次为:p H值料液比提取时间超声功率。与传统提取方法相比,超声辅助法使得蛋白提取量增加了0.81 g/(100 g)(占传统方法提取量的15.46%),蛋白纯度提高了5.47%。     

Studies on defatted seed removal efficiency for organochlorine compounds

Defatted seeds were evaluated for effective adsorption of organochlorine compounds such as chloroform, dichloromethane, and trichloroethylene. The amounts of these compounds adsorbed were plotted against the equilibrium concentration of substances in solution on a logarithmic scale. A linear relationship was obtained, indicating that the adsorption reactions were of the Freundlich type. The removal of these organochlorine compounds by defatted seed was attributed to the uptake by intracellular particles called spherosomes.     

Soyasaponins resist extrusion cooking and are not degraded during gut passage in Atlantic salmon (Salmo salar L.)

The stability of soyasaponins in fish feed formulations was investigated. The level of soyasaponin Ab, Bb, Bc, Ba-2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one (Ba-DDMP), Bb-DDMP, and Bc-DDMP was quantified in 15 samples of defatted soybean meal, two full fat soybean meals, and two soybean protein concentrates by reverse phase high-performance liquid chromatography. The total level of saponins in the 15 samples of commercial defatted soybean meal ranged from 4.8-6.8 micromol/g (5.1-7.0 g/kg). The two full fat meals contained 4.4 and 4.7 micromol/g whereas no saponins could be detected in the alcohol-extracted soybean protein concentrates. Fifteen batches of fish feed containing 20% defatted soybean meal were produced by twin-screw extrusion from the 15 different samples of defatted soybean meal. Extrusion did not reduce the total level of group B saponins, but the ratio between DDMP-conjugated group B saponins and non-DDMP-conjugated group B saponins was slightly reduced. A soybean-containing diet was fed to seawater adapted Atlantic salmon for 9 weeks. Yttrium oxide was included in the feed as an inert marker in order to estimate the disappearance of saponins during gut passage. High levels of intact non-DDMP-conjugated group B soyasaponins were found in feces whereas only low levels of DDMP-conjugated saponins could be detected. The overall disappearance of saponins was close to zero, and the concentration of intact saponins in dry feces reached levels several fold higher than dietary levels. The present work demonstrates that non-DDMP-conjugated group B soyasaponins resist extrusion cooking and remain intact during gut passage in Atlantic salmon. The latter is contrary to earlier findings in endothermic animals.     

Chemical composition of glyphosate-tolerant soybean 40-3-2 grown in Europe remains equivalent with that of conventional soybean (Glycine max L.)

The composition of glyphosate-tolerant (Roundup Ready) soybean 40-3-2 was compared with that of conventional soybean grown in Romania in 2005 as part of a comparative safety assessment program. Samples were collected from replicated field trials, and compositional analyses were performed to measure proximates (moisture, fat, ash, protein, and carbohydrates by calculation), fiber, amino acids, fatty acids, isoflavones, raffinose, stachyose, phytic acid, trypsin inhibitor, and lectin in grain as well as proximates and fiber in forage. The mean values for all biochemical components assessed for Roundup Ready soybean 40-30-2 were similar to those of the conventional control and were within the published range observed for commercial soybean. The compositional profile of Roundup Ready soybean 40-3-2 was also compared to that of conventional soybean varieties grown in Romania by calculating a 99% tolerance interval to describe compositional variability in the population of traditional soybean varieties already on the marketplace. These comparisons, together with the history of the safe use of soybean as a common component of animal feed and human food, lead to the conclusion that Roundup Ready soybean 40-3-2 is compositionally equivalent to and as safe and nutritious as conventional soybean varieties grown commercially.     

Comparison of nutritional and antinutritional factors in soybean and fababean seeds with or without cortex

The composition and contents of nutritional factors such as proteins, lipids, carbohydrates, fibers, amino acids, and antinutritional factors such as trypsin inhibitors, phytic acid, and tannins were compared in soybean and fababean seeds with emphasis placed on the nutritional improvement of the seeds by cortex removal. Protein hydrolysis analysis for both whole seeds and seed with cortex removed revealed the presence of a large amount of lysine, arginine, aspartic acid, glutamic acid, glycine, and leucine while these seeds contained a low level of tryptophan, cystine, and methionine. Some antinutritional factors such as trypsin inhibitors, phytic acid, and tannins were detected in soybean and fababean seeds: phytic acid content and trypsin inhibitor activity were higher in soybean seeds than in fababean seeds while the difference in the tannin content was less pronounced. It was found that most of the tannins occurred in the cortex of the soybean and fababean seeds. Tannins are polyphenolic compounds that readily form indigestible complexes with proteins and other macromolecules under specific environmental conditions. By removal of the cortex, tannins were almost completely eliminated without changes in the protein composition and amino acids. From these results, it is assumed that since soybean and fababean seeds contained a high concentration of antinutritional factors in the cortex such as tannins, the utilization of the legume seeds after removal of all of the cortex is suitable for human diet or industrial products.     

Study on furundu, a traditional Sudanese fermented roselle (Hibiscus sabdariffa L.) seed: effect on in vitro protein digestibility, chemical composition, and functional properties of the total proteins

Furundu, a meat substitute, is traditionally prepared by cooking the karkade (Hibiscus sabdariffa L.) seed and then fermenting it for 9 days. Physicochemical and functional properties of raw and cooked seed and of furundu ferments were analyzed. Furundu preparation resulted in significant changes in karkade seed major nutrients. Total polyphenols and phytic acid were also reduced. The increase in total acidity and fat acidity coupled with a decrease in pH indicates microbial hydrolysis of the major nutrients; proteins, carbohydrates, and fats. In vitro digestibility of the seed proteins reached the maximum value (82.7%) at the sixth day of fermentation, but thereafter it significantly decreased. The effect of furundu preparation on N solubility profiles and functional properties, such as emulsification and foaming properties and other related parameters, is investigated in water and in 1 M NaCl extracts from defatted flour samples. The results show that cooking followed by fermentation affects proteins solubility in water and 1 M NaCl. The foaming capacity (FC) from the flour of raw seed decreased as a result of cooking. Fermentation for 9 days significantly increased the FC of the cooked seed, restoring the inherent value. Foam from fermented samples collapsed more rapidly during a period of 120 min as compared to the foam from raw and cooked karkade seeds; stability in 1 M NaCl was lower as compared to those in water. In water, the emulsion stability (ES) from the fermented samples was significantly higher than that of the raw seed flour. Addition of 1 M NaCl significantly decreased the ES of the fermented samples.     

Antioxidant capacity of seed coat, dehulled bean, and whole black soybeans in relation to their distributions of total phenolics, phenolic acids, anthocyanins, and isoflavones

Black soybeans have been used as an excellent dietary source for disease prevention and health promotion in China for hundreds of years. However, information about the distribution of health-promoting phenolic compositions in different physical parts of black soybean and the contribution of phenolic compositions to overall antioxidant capacity is limited. To elucidate the distribution of phenolic composition and their contribution to antioxidant activities in black soybean, the total and individual phenolic profiles, and antioxidant capacities of seed coat, dehulled and whole black soybean were systematically investigated. The seed coat exhibited much higher total phenolic indexes and antioxidant activities than whole and dehulled black soybean. Dehulled black soybean possessed similar levels of total phenolic content, total flavonoid content, 2-diphenyl-1-picryhydrazyl (DPPH) radical scavenging activity, ferric reducing antioxidant power (FRAP), and oxygen radical absorbance capacity (ORAC) activities as compared to whole yellow soybean. Cyanidin-3-glucoside, petunidin-3-glucoside, and peonidin-3-glucoside were detected in the seed coat but not in dehulled black soybean and yellow soybean. Among benzoic acid detected, caffeic and chlorogenic acid were the predominant phenolic acids. Whole black soybean and dehulled black soybean exhibited similar isoflavone contents in 7- O-beta-glucosides and malonylglucosides of daidzein and genistein. The seed coat possessed significantly ( p < 0.05) lower 7- O-beta-glucosides and malonylglucosides of daidzein and genistein, acetylglycitin, and total isoflavones than whole and dehulled black soybean. The contribution of phenolics in the seed coat to the antioxidant activity of black soybean parts depends on the assay methods. When measured with the DPPH and FRAP methods, the seed coat contributed 90% of the total antioxidant capacity of black soybean. However, when measured with the ORAC method, the seed coat and dehulled portion contributed approximately equally the total antioxidant capacity of black soybeans. The information generated from this study on the distribution and content of their active components is useful for the effective use of black soybeans as an ingredient for promoting health.     

Analysis of isoflavones and phenolic compounds in Korean soybean [Glycine max (L.) Merrill] seeds of different seed weights

The seeds of 322 Korean soybean varieties were collected from six different cultivated sites in Korea and classified into three groups based on the 100-seed weight as small, medium, and large. Seeds were analyzed for their concentrations of isoflavones and phenolic compounds. The total average isoflavones in soybean cultivated at Iksan (2.840 micromol g(-1)) and phenolic compounds in soybean grown at Yeoncheon (9.216 micromol g(-1)) and Iksan (9.154 micromol g(-1)) were significantly different (p<0.05). In small and medium seeds of soybeans cultivated at Yeoncheon, Yesan, and Milyang high levels of isoflavones were obtained, whereas soybeans grown in Chuncheon showed the lowest isoflavone concentrations. However, isoflavone concentrations in the large seeds of soybean cultivated at Chuncheon showed the highest level. The soybean cultivated at Yeoncheon had high levels of phenolic compounds in small, medium, and large seeds, whereas the soybean grown at Chuncheon had the lowest. On the other hand, the phenolic concentrations of large soybean cultivated at Milyang were the least. At Yeoncheon, Yesan, and Milyang, the total isoflavone and phenolic compounds levels related to their seed size was significantly different (p<0.05), whereas in the soybean of different sizes cultivated at Chuncheon, the relationship to their seed size was not significantly different. The relationships of total isoflavones and phenolic compounds of small and medium soybean seeds were significantly higher than that of large soybean seeds. The hydroxybenzoic acid group in all sizes of seeds cultivated at six sites in Korea was the major phenolic compound, followed by flavonoid and hydroxycinnamic acid. The total isoflavone concentration was positively correlated with acetylglycoside and negatively correlated with malonylglycoside in the small soybean seeds cultivated at Yeoncheon. In medium soybean seeds cultivated at Yeoncheon, a significantly positive correlation was found between acetylglycoside and glycoside, between aglycone and glycoside, and between aglycone and acetylglycoside, whereas a significantly negative correlation was shown between malonylglycoside and glycoside, between acetylglycoside and malonylglycoside, and between aglycone and malonylglycoside. In large soybean seeds cultivated at Chuncheon, significantly positive and negative correlations were similar to those of medium seeds. The results presented here can improve the understanding of the relationships among the concentrations of individual chemical compounds and each chemical compound group and total chemical compounds in soybeans of different seed sizes from different cultivated sites.     

大豆籽粒的化学-力学特性灰色关联度及本构模拟

为研究大豆籽粒在不同受载情况下的力学特性,探索其化学-力学特性之间的关系,该文选择11种大豆籽粒在含水率为8.65%下对其进行化学组分、针尖压入、剪切、压缩试验,并借助Abaqus软件建立本构模型,对压缩试验过程进行模拟仿真。力学试验和模拟结果表明:所测大豆籽粒硬度为18.39~52.58 N/mm,大豆籽粒破损强度为3.65~15.32 MPa,大豆籽粒极限剪切力为12.70~52.33 N,纵轴的抗剪能力明显高于横轴;不同压缩形式和剪切方向分别对大豆籽粒破损强度和极限剪切力影响极显著;试验与仿真的载荷-变形曲线拟合良好,说明所建立的大豆本构模型能够分析研究其抗挤压特性。灰色关联度分析结果表明:与化学组分含量最为密切的力学指标是硬度与接触刚度,其中硬度与粗蛋白质、粗脂肪、粗淀粉、粗纤维含量的关联度分别为0.309 4、0.327 8、0.171 9、0.191 8,接触刚度与其关联度分别为0.220 6、0.283 7、0.186 9、0.133 4,粗蛋白质和粗脂肪含量对硬度与接触刚度的影响最大,其次是粗纤维素,粗淀粉含量。研究结果可为品质预测和品种鉴别提供新的方法和依据。     

基于高光谱图像处理的大豆品种识别(英文)

大豆组分(油,蛋白质,脂肪等)在不同的大豆品种间差异很大。对于提高大豆品质来说,大豆品种识别是一个关键因素。该文利用高光谱图像技术对不同的大豆品种进行识别。利用高光谱成像系统获取大豆样本1 000～2 500 nm范围的光谱反射数据;应用主成分分析法(PCA,principal component analysis)对获取到的光谱数据进行数据降维并去除冗余数据;在分类算法中将得分高的主成分值作为输入特征,通过PCA方法从每个特征图像中提取4个特征变量(能量、熵、惯性矩和相关性);对于具体特征提取,从16个特征变量中提取8个重要特征参数;根据选择的特征,应用神经网络方法构建分类器;训练精度精度达到97.50%,平均测试精度达到93.88%以上。结果表明,应用高光谱图像技术结合神将网络建模方法可以对大豆品种进行分类。     

电喷雾萃取及电离质谱法分析大豆水提液快速鉴别种子活力

为快速评价大豆种子活力,该文以人工加速老化后的大豆种子为原料,采用电喷雾萃取电离质谱（extractive electrospray ionization-mass spectrometry,EESI-MS）直接检测不同活力大豆种子水提液,获得其正离子模式下指纹图谱,结合主成分分析（principal component analysis,PCA）、聚类分析（cluster analysis,CA）和判别分析（discriminant analysis, DA）,建立鉴别大豆种子活力的方法。结果表明,在正离子模式下,电喷雾萃取电离质谱结合多变量分析方法可以有效鉴别不同活力大豆种子。主成分分析提取了前3个主成分因子,累计贡献率达到94.80%,聚类分析和判别分析的正确率都为100%,对外部验证样本进行判别分析,正确率为95%。电喷雾萃取电离质谱能快速鉴别不同活力大豆种子,为种子活力检测提供了一种快速、准确、高效的新方法。     

Enzymatic Treatment of Soils Contaminated with Phenol and Chlorophenols Using Soybean Seed Hulls

This study presents the application of soybean seed hulls for the decontamination and/or detoxification of phenol and chlorophenol polluted soils. The effect of soil was examined on both catalytic activity of soybean seed hull peroxidase (SBP) and enzymatic transformation of phenol, 2-chlorophenol, and 2,4-dichlorophenol through the polymerization reaction. The sorption of the enzyme to soil organic matter was found to account for the loss of partial catalytic activity of soybean seed hulls in a soil slurry environment. The organic matter present in loamy soils, rather than other soluble soil constituents and soil micro-organisms, is a factor in SBP inhibition by soil and the corresponding decline in treatment efficiency of phenol and chlorophenols. Under improved conditions, however, soybean seed hulls demonstrated a satisfactory ability to catalyze the polymerization reaction. Over 96% of total phenols were removed using soybean seed hulls in a soil slurry bioreactor, which demonstrates a great potential in use of soybean seed hulls, a readily available and inexpensive source of SBP, for bioremediation of soils contaminated with phenolic compounds.     

Porphyran as a functional modifier of a soybean protein isolate through conjugation by the Maillard reaction

Porphyran (Por) prepared from dried nori was applied as a functional modifier of a soybean protein isolate (SPI) to conjugate with SPI from defatted soybean by the Maillard reaction (79% relative humidity and 60 degrees C for 7 days). Two kinds of partially denatured conjugate (Conj 45 and Conj 63) were obtained from the reaction product by sequential extraction at pH 4.5 and pH 6.3, and the respective yield and weight ratios of the SPI and Por moieties were 8.4% and 1:1 for Conj 45 and 11.7% and 1:0.16 for Conj 63. Conj 63 demonstrated improved solubility between pH 5.0 and pH 8.0, while Conj 45 exhibited substantially complete solubility over the pH range of 2.0-8.0. Conj 63 showed more tolerance against digestion with pancreatin than SPI, whereas this was lost after denaturation. Conj 63 and Conj 45 both showed a markedly higher emulsion activity index and emulsion stability than SPI, even at pH 3.0; in particular, Conj 45 exhibited outstanding emulsifying ability. Conj 63 had about a two-fold higher calcium-binding ability than SPI, and Conj 63 and Conj 45 did not aggregate with added Ca2+ and Mg2+. It is believed that Por could be a valuable functional modifier of SPI for providing soybean protein-based liquid foods such as beverages by conjugation through the Maillard reaction.     

Positive correlation between the number of root nodule primordia and seed sugar secretion in soybean (Glycine max L.) seedlings inoculated with a low density of Bradyrhizobium japonicum

Sufficient rhizobium population in the rhizosphere of legume seedlings is required for early and enough setting of root nodules. Potential of seed and seedling root exudates for proliferation of Bradyrhizobium japonicum PNT119 was evaluated in the soybean cvs. Enrei and Tachinagaha. In both cultivars, seed exudates showed a higher potential than root exudates. In the seed exudates, a low-molecular high-polarity fraction including sugars and amino acids showed a higher potential than both high-molecular fraction and low-molecular low-polarity fraction. The correlations between the sugar or amino acid contents of seed exudates and the number of root nodule primordia at 7 d after sowing were investigated among 12 soybean cultivars. When seeds were inoculated with a low rhizobium density, a high correlation coefficient was detected between them. However, there was no positive correlation in the plants inoculated with a high rhizobium density. These results suggest that the amount of low molecular substances in the seed exudates determines the number of root nodule primordia through rhizobium proliferation around seed and young roots when the rhizobium density is the limiting factor. The genetic trait relating to seed coat secretion should be considered as a possible key factor contributing to adequate root nodulation in soybean seedlings cultivated in fields with a low rhizobium density.     

Fermentation as a bio-process to obtain functional soybean flours

The effect of fermentation on the antioxidant compounds [vitamins C and E, total phenolic compounds (TPC), and reduced glutathione (GSH)], and antioxidant capacity [superoxide anion scavenging activity (SOD-like activity), peroxyl radical-trapping capacity (PRTC), inhibition of phosphatidylcholine (PC) peroxidation, and Trolox equivalent antioxidant capacity (TEAC)] of soybean (Glycine max cv. Merit) was studied. Fermentation was carried out in solid state in cracked seeds inoculated with Aspergillus oryzae, Rhizopus oryzae, Bacillus subtilis, and Lactobacillus plantarum and in liquid state either in cracked seeds or milled soybean flours fermented naturally by only the microorganisms present in the seeds or by inoculation with L. plantarum. Vitamin C was not detected in the studied samples. Fermentation caused a decrease in vitamin E activity, except when cracked seed was fermented with A. oryzae, R. oryzae, or B. subtilis that increased 31, 30, and 89%, respectively. Fermentation produced an increase in TPC content and did not affect or reduce the GSH content. Fermentation decreased SOD-like activity drastically, while PRTC increased except when it was carried out naturally in cracked seed. TEAC values rose sharply when soybeans were fermented with B. subtilis. Processed soybean extracts inhibited PC peroxidation in comparison with the control assay. On the basis of the results obtained, the relative contributions of vitamin E, TPC, and GSH to antioxidant capacity were calculated and results showed a very high TPC contribution and a low contribution of GSH and vitamin E activity. Optimum results for functional soybean flours were achieved when fermentation was carried out with B. subtilis inoculum.