Sicklepod (Senna obtusifolia) seed processing and potential utilization

Sicklepod (Senna obtusifolia) is a leguminous plant that infests soybean fields in the southeastern United States. Its seeds contain a variety of toxic, highly colored compounds, mainly anthraquinones together with a small amount of fat. These compounds contaminate and lower the quality of soybean oil when inadequately cleaned soybean seed from this area is processed. The sorting of sicklepod seed from a soybean harvest is an additional economic burden on the farmer beyond the cost of proper disposal of the weed seed to avoid worsening field infestation. Fortunately, sicklepod seed also contains substantial amounts of carbohydrates and proteins. These edible components when freed from anthraquinones have a market in pet food as well as potential in human foods because of the high galactomannan ratio of the polysaccharides. Sicklepod seed was dehulled, and the ground endosperm was defatted, followed by sequential solvent extraction of the defatted seed meal to isolate the anthraquinones, carbohydrates, and protein components into their respective classes. Each class of isolate was spectroscopically identified.     

Effect of far-infrared irradiation on the antioxidant activity of defatted sesame meal extracts

To determine the effect of far-infrared (FIR) irradiation on the antioxidant activities of sesame meal, half of sesame seeds were FIR-irradiated and then oil was extracted from the seeds. The resulting defatted sesame meal (DSM) was extracted with methanol, and the antioxidant activities of methanolic extract were determined. FIR irradiation of sesame seeds for 30 min increased the total phenol content from 34.0 to 59.0 muM and radical scavenging activity of DSM extracts from 26.40 to 68.76%. The induction time of lipid oxidation of oil added to extracts was also retarded from 0.82 to 0.96 h. According to the gas chromatography-mass spectrometry analysis, several low molecular weight phenolic compounds, such as p-hydroxy benzoic acid, vanillic acid, p-coumaric acid, isoferulic acid, and o-coumaric acid, were frequently detected in FIR-irradiated DSM extracts as compared to unirradiated ones. These results indicated that FIR irradiation of sesame seeds increased the antioxidant activity of methanolic extracts of DSM.     

Enzymatic extractability of soybean meal proteins and carbohydrates: heat and humidity effects

To study the incomplete enzymatic extractability of proteins and carbohydrates of thermally treated soybean meals, one unheated and three heat-treated soybean meals were produced. To obtain truly enzyme-resistant material, the meals were extracted by a repeated hydrolysis procedure using excessive concentrations of different combinations of commercial protease and carbohydrase preparations. The water extractability of protein from the different meals varied considerably (13-67%). For all soybean meals, enzymatic treatment extracted most of the original protein (89-94%). Carbohydrase preparations did not improve protein extraction. High-humidity heat treatment led to a more effective enzymatic extraction, which seemed to correlate with the extent of protein denaturation. Results with purified proteins indicated that the soybean meal matrix affects the enzymatic extraction of protein from the meals. Interactions between protein and other components (e.g., cellulose) may explain the incomplete enzymatic extractability of protein from the meals.     

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.     

Aggregation of peptides during hydrolysis as a cause of reduced enzymatic extractability of soybean meal proteins

With the purpose of analyzing the size and composition of enzyme-unextractable proteins in differently heat-treated soybean meals, a selection of extractants was screened for their ability to extract these proteins from enzyme-unextractable residues. The largest effects were obtained with urea, urea plus beta-mercaptoethanol, and dilute alkali; the latter extracted up to 87% of the enzyme-unextractable protein. Gel permeation chromatography indicated that a large proportion of the extracted material was of high molecular weight. However, the combined results from gel electrophoresis, LC-MS, and MALDI-ToF MS showed that the extracted protein material was composed of aggregated peptides. The largest aggregates were observed in the enzymatic residues originating from meals heat-treated at high humidity. Extracted aggregates were fully degraded upon subsequent proteolytic treatment.     

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。     

Study of Three-Dimensional Structure of Wheat Starch Granules Stained with Remazolbrilliant Blue Dye and Extracted with Aqueous Sodium Dodecyl Sulfate and Mercaptoethanol Solution

Wheat starch granules were obtained from soft wheat flour by acetic acid fractionation (pH 3.5), and the starch was stained by reaction with Remazolbrilliant blue (RBB) dye. RBB-stained starch was extracted with 1% sodium dodecyl sulfate (SDS) and 1% 2-mercaptoethanol (ME) for 14.5 hr at room temperature. This extraction step was repeated five times (extracts 1–5). SDS-ME extracts were subjected to size-exclusion column chromatography, and comparisons of their profiles for specific absorbance at 650 nm (A₆₅₀) and carbohydrates were made. After high molecular weight (HMW) carbohydrates on the starch granule surface were extracted, HMW carbohydrates inside the granule appeared to be extracted. Finally, low molecular weight (LMW) carbohydrates near the granule surface were extracted. Phase-contrast light microscopy of the treated starch granules showed that all granules became transparent. Two different interior structures were observed. Scanning electron microscopy indicated that the granule was split into two parts at the equatorial groove. The interior of the granule showed two different areas: a central hole area and the surrounding stratified area. Extraction beyond five times with the same solvent dissolved the weak part of the granule structure and left two types of skeletal structures. The appearance of the skeletal structure of the granule surface was different from the appearance of interior structures.     

Selective one-step extraction of Arabidopsis thaliana seed oleosins using organic solvents

Oleosins are hydrophobic proteins from oleaginous seeds, surrounding and stabilizing oil bodies. They are known to display interesting interfacial properties. Specific sera were raised against four different A. thaliana oleosins and used in dot-blot assays for oleosin quantification. These assays were used to set up extraction of oleosins from A. thaliana seeds. One mixture of chloroform/methanol gave optimal oleosin extraction. Extracted proteins represented 9% of seed proteins and were identified by immunoblot and proteomic analyses. Oleosins accounted for 79% of the extracted proteins. This simple one-step procedure allows selective extraction and concentration of oleosins from seeds without tedious oil body purification. Oleosin extract was indeed used to demonstrate the presence of the rare oleosin S5 in mature seeds. Moreover, this method will be useful to investigate the potential use of oleosins as emulsifier and to question their possible allergenicity.     

Lunasin and Bowman-Birk protease inhibitor concentrations of protein extracts from enzyme-assisted aqueous extraction of soybeans

Lunasin and Bowman-Birk protease inhibitor (BBI) are two soybean peptides to which health-promoting properties have been attributed. Concentrations of these peptides were determined in skim fractions produced by enzyme-assisted aqueous extraction processing (EAEP) of extruded full-fat soybean flakes (an alternative to extracting oil from soybeans with hexane) and compared with similar extracts from hexane-defatted soybean meal. Oil and protein were extracted by using countercurrent two-stage EAEP of soybeans at 1:6 solids-to-liquid ratio, 50 °C, pH 9.0, and 120 rpm for 1 h. Protein-rich skim fractions were produced from extruded full-fat soybean flakes using different enzyme strategies in EAEP: 0.5% protease (wt/g extruded flakes) used in both extraction stages; 0.5% protease used only in the second extraction stage; no enzyme used in either extraction stage. Countercurrent two-stage protein extraction of air-desolventized, hexane-defatted soybean flakes was used as a control. Protein extraction yields increased from 66% to 89-96% when using countercurrent two-stage EAEP with extruded full-fat flakes compared to 85% when using countercurrent two-stage protein extraction of air-desolventized, hexane-defatted soybean flakes. Extruding full-fat soybean flakes reduced BBI activity. Enzymatic hydrolysis reduced BBI contents of EAEP skims. Lunasin, however, was more resistant to both enzymatic hydrolysis and heat denaturation. Although using enzymes in both EAEP extraction stages yielded the highest protein and oil extractions, reducing enzyme use to only the second stage preserved much of the BBI and Lunasin.     

Overexpression, purification, and in vitro refolding of the 11S globulin from amaranth seed in Escherichia coli

An amarantin 11S globulin cDNA encoding one of the most important storage proteins of amaranth seeds, with a high content of essential amino acids, was expressed in Escherichia coli. A good level of expression of recombinant amarantin with a molecular weight of 59 kDa was obtained. The recombinant protein was extracted by ammonium sulfate precipitation and purified to homogeneity using ion-exchange chromatography and reversed phase high-performance liquid chromatography. The expressed protein exhibited electrophoretic, immunochemical, and surface hydrophobicity properties similar to those of native amarantin from amaranth seed. Also, the recombinant protein was refolded in vitro using two different methods.     

Investigation and optimization of the factors influencing sorghum protein extraction

To optimize the extraction of sorghum proteins, several variables were examined: sample-to-solvent ratio, detergent type and concentration, reducing agent type and concentration, extraction time, and buffer pH and concentration. Samples were quantified and characterized by RP-HPLC, FZCE, and nitrogen analysis. These studies revealed that pH, detergent type, reducing agent type, and sample-to-solvent ratio all had significant effects on the levels of protein extracted. Increasing SDS concentration (2%) and solvent-to-flour ratio (20:1) with multiple 5 min extracts reduced extraction time by 35-80% while still extracting the same levels of total protein relative to the control methodology. Reproducibility using the multiple extractions was found to be excellent with relative standard deviations of <2% for consecutive extractions.     

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.     

Real-time polymerase chain reaction quantification of the transgenes for roundup ready corn and roundup ready soybean in soil samples

A method for quantification of recombinant DNA for Roundup Ready (RR) corn and RR soybean in soil samples is described. Soil DNA from experimental field samples was extracted using a soil DNA extraction kit with a modified protocol. For the detection and quantification of recombinant DNA of RR corn and RR soybean, a molecular beacon and two pairs of specific primers were designed to differentially target recombinant DNA in these two genetically modified crops. Soil DNA extracts were spiked with RR corn or RR soybean DNA, and recombinant DNA was quantified using real-time PCR with a molecular beacon. As few as one copy of RR corn genome or one copy of RR soybean genome was detected in the soil DNA extract.     

Hydroponic cultivation improves the nutritional quality of soybean and its products

Hydroponic cultivation allows the control of environmental conditions, saves irrigation water, increases productivity, and prevents plant infections. The use of this technique for large commodities such as soybean is not a relevant issue on fertile soils, but hydroponic soybean cultivation could provide proteins and oil in adverse environmental conditions. In this paper, the compositions of four cultivars of soybean seeds and their derivates, soy milk and okara, grown hydroponically were compared to that of the same cultivar obtained from soil cultivation in an open field. Besides proximal composition, the concentrations of phytic acid and isoflavones were monitored in the seeds, soy milk, and okara. Results demonstrated that, independent from the cultivar, hydroponic compared to soil cultivation promoted the accumulation of fats (from 17.37 to 21.94 g/100 g dry matter) and total dietary fiber (from 21.67 to 28.46 g/100 g dry matter) and reduced isoflavones concentration (from 17.04 to 7.66 mg/kg dry matter), whereas protein concentration was unaffected. The differences found in seed composition were confirmed in the respective okara products, but the effect of cultivation system was not significant looking at the soy milk composition. Data showed that hydroponic cultivation improved the nutritional quality of soybean seeds with regard to fats and dietary fiber. They also suggest that specific cultivars should be selected to obtain the desired nutritional features of the soybean raw material depending on its final destination.     

基于OpenCV的大豆籽粒多表型参数获取算法

大豆籽粒的表型参数获取对大豆育种具有重要的作用。现有的深度学习算法获取的大豆籽粒表型性状较少，且识别表型的神经网络模型训练成本高。本研究基于OpenCV图像处理库，提出了一种提取大豆籽粒多表型参数的算法，从大豆图像中一次性获取籽粒的多种表型性状参数，同时能识别大豆的优劣品质。将每个待测大豆单株的所有籽粒拍成一张图像，首先对大豆籽粒图像进行二值化、去噪等预处理，然后采用分水岭算法和改进的目标分割算法提取图像中的大豆籽粒轮廓。根据大豆籽粒的轮廓信息，调用OpenCV图像处理函数计算大豆籽粒的个数、长轴长度、短轴长度、面积、周长等多个表型性状参数。引入圆形度识别残缺大豆籽粒，使用RGB阈值判断识别病变大豆籽粒。测试结果表明，采用本文算法计算的颗粒总数识别率为98.4%，大豆籽粒正确识别率为95.2%，破损大豆和病变大豆的识别率分别为91.25%和88.94%，籽粒的长轴长度与短轴长度的测量精度分别为96.8%、95.8%；引入多进程并行计算，该算法的 处理215张图片时间为248.9 s，相对于单进程计算缩短了约2/3，实现了低成本高通量的高精度大豆籽粒多表型性状参数的自动获取，为大豆籽粒自动化考种提供有效的处理方法。     

Role of sulfur nutrition in plant and seed metabolism of Glycine max L.

The aim of present investigation has been to explore the effect of sulfur application on plant metabolism, seed yield and seed quality in soybean. The sulfur was supplied in different doses ranging as 1, 2, 4, 6 and 8 meq S L^?1. Plant supplied with 4 meq S L^?1 showed optimal growth. Plant growth and dry matter was reduced under sulfur deficiency (1 and 2 meq S L^?1) and toxicity (6 and 8 meq S L^?1). Application of sulfur increases the tissue sulfur and cysteine concentration in both leaves and seeds. The critical concentration for deficiency (CCD) and toxicity (CCT) of sulfur was observed 0.194 to 0.277% dry weight respectively. Pod yield and seed yield was also suppressed in sulfur deficiency and toxicity. In leaves sugar (reducing, non-reducing and total sugar) and starch was found to be accumulated while in seeds both were depleted under sulfur deficiency and toxicty. Seed storage proteins (albumins, globulins, glutelins and prolamins) were also reduced under sulfur stress. Thus, we conclude that sulfur deficiency and toxicity both affects the plant metabolism, yield and seed quality in terms of carbohydrates and storage proteins of soybean.     

Bioactive peptides in amaranth (Amaranthus hypochondriacus) seed

Amaranth seeds are rich in protein with a high nutritional value, but little is known about their bioactive compounds that could benefit health. The objectives of this research were to investigate the presence, characterization, and the anticarcinogenic properties of the peptide lunasin in amaranth seeds. Furthermore, to predict and identify other peptides in amaranth seed with potential biological activities. ELISA showed an average concentration of 11.1 microg lunasin equivalent/g total extracted protein in four genotypes of mature amaranth seeds. Glutelin fraction had the highest lunasin concentration (3.0 microg/g). Lunasin was also identified in albumin, prolamin and globulin amaranth protein fractions and even in popped amaranth seeds. Western blot analysis revealed a band at 18.5 kDa, and MALDI-TOF analysis showed that this peptide matched more than 60% of the soybean lunasin peptide sequence. Glutelin extracts digested with trypsin, showed the induction of apoptosis against HeLa cells. Prediction of other bioactive peptides in amaranth globulins and glutelins were mainly antihypertensive. This is the first study that reports the presence of a lunasin-like peptide and other potentially bioactive peptides in amaranth protein fractions.     

Imbibition of soybean seeds in warm water results in the release of copious amounts of Bowman-Birk protease inhibitor, a putative anticarcinogenic agent

Protease inhibitors play a protective role against pathogenic microorganisms and herbivorous insects. The two predominant protease inhibitors of soybean seeds are the Kunitz trypsin inhibitor (KTI) and Bowman-Birk protease inhibitor (BBI). In this study, we report that soybean seeds incubated in warm water release large amounts of proteins into the surrounding media. Two-dimensional gel electrophoresis analysis of the seed exudates resulted in the separation of 93 distinct protein spots out of which 90 spots were identified by LC-MS/MS. The basic 7S globulin and the BBI are the two predominant proteins found in the soybean seed exudates. In addition to 7S and 11S seed storage proteins, others known to protect the seeds against pathogens and pests including KTI, peroxidase, α-galactosidase, and endo-1.3-β-glucanase were also identified in the seed exudates. Soybean seed exudate obtained by incubating the seeds in warm water was also able to inhibit the growth of human breast cancer cell line MCF-7. Since soybean seeds release large amounts of enzymatically active BBI when immersed in warm water, our procedure could be exploited as a simplified alternative method for the preparation of BBI concentrate which is being used as a cancer chemoprotective agent.     

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.     

葡萄籽中水溶性钙结合蛋白的分离和鉴定

为了开发植物源葡萄籽补钙制剂,该研究通过亚细胞定位试验表明,葡萄籽的胚乳中含有大量的钙元素。通过电泳分析发现,葡萄籽的水溶性蛋白包括2种主要成分,其中一种是11 S球蛋白(蛋白质B),也是最主要的钙结合蛋白,另一种是表观分子量为670 k Da的蛋白质A。在蛋白质组成相同的情况下,用传统的碱溶酸沉法来分离葡萄籽蛋白会导致大量的钙流失。但用30%~50%硫酸铵沉淀法得到的蛋白质得率是(22.5±0.02)g/kg,蛋白质中钙质量分数(3.47%)是碱溶酸沉法(1.11%)的3倍。该研究结果为食品工业中矿物结合蛋白质的分离纯化提供参考。