Nutritional assessment of seed proteins in rice bean [Vigna Umbellata (Thumb.) Ohwi and Ohashi

Seeds of three rice bean accessions had 17.26 to 21.42% protein, 3.46 to 4.03% fat, 61.09 to 64.73% carbohydrates 3.99 to 4.58% ash and 5.22 to 7.43% fiber (dry weight basis).The most limiting amino acids in the seed meal, albumin and globulin fractions, were methionine and cysteine with chemical scores of these fractions being 38% to 59%. The amino acid pattern of globulin and seed meal were similar.Thein vitro protein digestibility (IVPD) ranged from 82 to 86% for the seed meal, 86 to 88.5% for the albumin and 75.9 to 83.3% for the globulin. Relative nutritive values (RNV) of raw mature seed of two accessions were 22.6% and 42.4% and increased to 55.6% to 79.4% after boiling and roasting.Part of MS thesis of the senior author.     

Distribution of seed protein fractions and amino acids in different anatomical parts of chickpea (Cicer arietinum L.) and pigeonpea (Cajanus cajan L.)

Studies on protein fractionation in seed coat, embryo, cotyledons and whole seed were made to observe the differences, if any, between chickpea and pigeonpea. Results indicated that globulin was the major fraction of embryo and cotyledons of these legumes. Seed-coat nitrogen was observed to be mostly comprised of nonprotein nitrogen and glutelin fractions and thus differed from other components in both chickpea and pigeonpea. The albumin fraction of cotyledons of both crops had the highest concentration of sulphur amino acids, methionine and cystine. Glutelin contained a considerably higher concentration of methionine and cystine than did globulin in chickpea and pigeonpea. This suggests that lines with higher glutelin should be identified to improve their protein quality. The amino acid compositions of different seed components did not show large differences between these two pulse crops.Submitted as JA no. 180 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT).     

Chemical analysis and nutritional assessment of the less known pulses,Vigna aconitifolia (Jacq.) Marechal andVigna vexillata (L.) A. Rich.

The seeds ofVigna aconitifolia andVigna vexillata were analysed for proximate composition, minerals, seed protein fractions, amino acids, fatty acids, and antinutritional factors. Both the pulses were found to be rich sources of proteins and minerals like Ca, Mg, Fe, Zn, and Mn.Vigna aconitifolia seeds exhibited fairly high levels of crude lipid. The most limiting essential amino acids in both the pulses were the sulphur-amino acids, cystine, and methionine. Threonine inV. aconitifolia and phenylalanine and tyrosine inV. vexillata occurred in higher quantities when compared with WHO/FAO requirement pattern. Oleic acid and palmitic acid inV. aconitifolia and linoleic acid and palmitic acid inV. vexillata lipids were found to be the predominant fatty acids. The tannin content of both the pulses was negligible compared with the domesticated legumes. The other antinutritional factors like total free phenols, L-DOPA and haemagglutinating activity were also analysed/assayed.     

Fundamental study on protein changes taking place during malting of oats

During the malting process, storage proteins are degraded by proteolytic enzymes into small peptides and amino acids. The activity of these enzymes was measured during malting of oats and was found to be increased. To quantify proteolytic degradation, proteins of unmalted, germinating and malted grains were fractionated. After extracting the oat proteins (Osborne fractionation), protein fractions were analysed using a Lab-on-a-Chip technique, which separates the proteins – based on their molecular weight – by capillary electrophoresis. This new technique for the analysis of proteins was supported by using two-dimensional gel electrophoresis. In addition, amino acid analysis was carried out. In general a degradation of proteins to small peptides and amino acids could be observed in the globulin, prolamin and glutelin fractions. In the albumin fraction a protein increase was observed, which is due to the fact that this fraction contains the majority of the metabolically active proteins. Amino acid analysis supported the observation of increased protein amount in the albumin fraction and decreased protein amounts in the other fractions. Some proteins, which have not been described in the literature, were detected in the albumin and glutelin fraction, since Lab-on-a-Chip technique allows detection of proteins with low molecular weights of 4.5 kDa.     

Changes in nitrogenous and other chemical constituents,protein fractions and in vitro protein digestibility of germinating fluted pumpkin ( Telfairia occidentalis Hook) seed

The effect of 7 days of germination on levels of nitrogenous and other nutrition related parameters, protein fractions and in vitro protein digestibility of fluted pumpkin (Telfairia occidentalis) seed was studied. The non-protein nitrogen gradually increased and the protein nitrogen content decreased during germination. Albumin and globulin fractions were found to be the major seed proteins of fluted pumpkin seeds, constituting about 58.6% of the total protein of the ungerminated (raw) seeds. The protein fractions, albumin and glutelin, were observed to increase by 61.5% and 57.0%, respectively, while a 54.6% decrease was noted in the prolamine fraction. The globulin fraction increased at the beginning of germination but decreased at the end. Germination significantly (p 0.05) increased the crude protein, nitrogen solubility and in vitro protein digestibility but decreased the fat, phytic acid and polyphenol contents of the seeds.     

Characterisation and functional properties of Australian rice protein isolates

Albumin, globulin, glutelin and prolamin fractions were isolated from an Australian rice variety (cv. Langi) and characterised by yield, protein content and molecular weight profile using both capillary electrophoresis (SDS-CE) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The influence of pre-extraction enzymatic hydrolysis of starch and heating to 70 °C was also investigated, as was the extraction of the glutelin fraction without prior removal of the albumin and globulin fractions. Pre-extraction treatment affected mainly the albumin fraction, increasing dry matter yield but reducing protein content. SDS-CE was able to separate the protein fractions over a wider molecular weight range than SDS-PAGE, and the peaks from SDS-CE showed slightly higher molecular weight compared to equivalent bands from SDS-PAGE. The glutelin fraction extracted without prior removal of albumin and globulin fractions had different characteristics compared to those obtained by conventional extraction methods. Pre-extraction hydrolysis of starch did not significantly affect the emulsifying, foaming and gelling properties of extracted protein. Although rice glutelin had poor solubility, emulsifying and foaming properties in aqueous systems, it had good gelling properties which could be important for food applications.     

Extractability and chromatographic separation of rice endosperm proteins

The molecular weight (MW) distribution of proteins extracted with different solvents from defatted rice endosperm was examined by size exclusion-high performance liquid chromatography (SE-HPLC) with 2.0% sodium dodecyl sulfate (SDS) (w/v) as mobile phase. The resulting protein peaks were further characterized by SDS-PAGE. Under the experimental conditions, 2.0% SDS extracted 64% of the proteins. Adding 6.0 M urea resulted in a 15% increase in extractability (up to 79%). With using 20–100 mM NaOH, 70–81% of the proteins were extractable. Maximum extractability was reached with 2.0% SDS, 6.0 M urea and 0.5–1.5% dithiothreitol (DTT). Apparent MW profiles of rice endosperm proteins allowed classification into six fractions of decreasing apparent MW. Fraction VI contained the low MW albumin, globulin, and prolamin protein material. Fractions IV and V originated from α and β glutelin subunits, respectively. The polypeptides of fraction III consisted of an α and a β subunit linked by an intermolecular disulfide bond. The polypeptides of fractions I and II were dimers, trimers or more highly polymerized forms of the (α–β) glutelin subunit dimer in fraction III. While the work confirmed that rice glutelin is composed of polymers of α and β subunits, remarkably, higher MW glutelin aggregates (fractions I–III) only partly dissociated on reduction. Low MW protein material (fraction VI) was entrapped in the aggregated protein network and was released on reduction. The rapid and reproducible SE-HPLC method developed for rice protein separation allows a more quantitative approach than SDS-PAGE.     

Characterization and In Vitro digestibility of rice protein prepared by enzyme-assisted microfluidization: Comparison to alkaline extraction

Microfluidization followed by density-based separation was employed to extract protein from broken rice by disrupting protein-starch agglomerates. Follow-up enzyme treatments (amylase and glucoamylase) were performed to further improve the purity of the protein-rich fraction. High protein recovery (81.87%) and purity (87.89%) were obtained. The protein composition, solubility, structural properties, and in vitro digestibility of rice proteins prepared by enzyme-assisted microfluidization (EM-RP) and alkaline extraction (AE-RP) were compared. EM-RP was mainly composed of glutelin, which had low solubility and native structure. By contrast, large quantities of prolamin and globulin appeared in the AE-RP except glutelin, leading to the richness of glutamic acid/glutamine, leucine, aromatic and charged amino acids in the AE-RP. Compared to AE-RP, EM-RP showed higher digestibility due to the richness of glutelin (an easy-to-digest protein), as evidenced by higher nitrogen release during pepsin-trypsin digestion. The presence of prolamin (an indigestible protein) in AE-RP decreased protein digestibility although alkaline extraction improved its hydrolysis. These results suggest that enzyme-assisted microfluidization could be an effective technique to non-destructively and selectively extract rice glutelin.     

醋制对黑豆蛋白含量及多肽ACE抑制活性的影响

为考察醋制对黑豆蛋白含量及多肽ACE抑制活性的影响,为对黑豆醋浸过程中蛋白组分及含量变化进行分析,并对相应ACE抑制肽活性进行测定。黑豆经醋浸不同时间后,采用顺序抽提法提取清蛋白、球蛋白、醇溶蛋白和谷蛋白,并采用凯氏定氮法及SDS-PAGE电泳进行定量和定性分析;各蛋白组分经胃蛋白酶水解,经超滤(截留分子量3 k D)后收集滤液,获得多肽组分,RP-HPLC法进行ACE抑制活性评价。结果表明:经醋浸14 d后,黑豆总蛋白含量变化幅度小于±0.5%;清蛋白含量由55.13%(0 d)降低至9.61%(14 d);球蛋白由7.04%(0 d)增加到20.34%(14 d);醇溶蛋白由0.81%(0 d)增加到1.72(14 d);谷蛋白由21.11%(0 d)增加到59.45%(14 d),含量最高。醋浸处理降低了清蛋白源多肽的ACE抑制活性,但提高了球蛋白、醇溶蛋白及谷蛋白多肽的ACE抑制活性,其中,谷蛋白多肽抑制率由18.18%(0 d)增加至37.58%(14 d),抑制活性升高。醋浸可改变黑豆各蛋白组分的含量和对应多肽的ACE抑制活性,其中,谷蛋白含量及其多肽ACE抑制活性均有增加。研究结果表明可进一步采用分离纯化技术从谷蛋白多肽中获得高活性降压肽。     

我国江浙地区粳稻品种间的植酸与蛋白质组分差异及其相关性

选用我国江浙地区近期育成的晚粳稻品种（系）29个,对它们在浙江杭州相同栽培条件下籽粒中的植酸、蛋白质含量与其组分差异及其相关性进行了分析。供试29个品种(系)植酸含量的平均值为0.868%,变化幅度为0.699%~1034%,其中秀水系列品种的植酸含量一般较低,而武育粳系列和淮字系列品种（系）的植酸含量则相对较高;供试29个品种材料总蛋白质含量的平均值为8.722%,其含量大致呈正态分布。在4种蛋白质组分中,谷蛋白、球蛋白和清蛋白在品种间的变异系数较大,而醇溶蛋白则相对较小,且在分布频率上也表现出品种类型间的差异特征;除谷蛋白与总蛋白质之间存在着极显著的正相关外,供试29个品种(系)的植酸含量与蛋白质含量及4种蛋白质组分含量间并无密切的关系。     

Antioxidative properties of partially purified barley hordein, rice bran protein fractions and their hydrolysates

Antioxidative properties of proteins from barley and rice bran and their hydrolysates were examined. Three major hordein fractions of barley, B, C and D hordeins, were partially purified by gel filtration. Albumin, globulin, prolamin and glutelin fractions of rice bran were fractioned by the Osborne method. Hydrolysates of these protein fractions were prepared by digesting with pepsin followed by trypsin. Antioxidant properties in terms of antioxidative activity against linoleic acid peroxidation and reducing activity without the lipid adjuvant were investigated. The globulin fraction from rice bran protein revealed the strongest antioxidative activity throughout the incubation time of 7 days (p ≤ 0.05). The albumin fraction of rice bran protein showed the highest reducing activity (6964 μmol of Fe²⁺) followed by globulin, prolamin, glutelin and hordein fractions with activities of 2904, 2017, 1809 and 1333 μmol of Fe²⁺, respectively (p ≤ 0.05). Partially purified C hordein exhibited the highest reducing activity compared with B and D hordeins. Protein hydrolysates obtained after digestion with pepsin and trypsin exhibited much greater antioxidative, as well as reducing, activities than those from before digestion.     

Amino acid compositions of different protein fractions in developing grains of NP 113 barley and its high lysine Notch-2 mutant

The percent distributions of protein fractions namely albumin + globulin, prolamine and glutelin were studied in developing grains of NP 113 barley and its high lysine mutant Notch-2. During development the percentage of albumin + globulin fraction decreased in NP 113, while those of prolamine and glutelin remained unchanged. The increase in prolamine was substantial from 24 to 31DAA. In Notch-2 the trend followed by albumin + globulin and prolamine was like that in NP 113, while the glutelin fraction showed an increase as compared to 10 DAA. The percent of albumin + globulin was slightly higher in Notch-2 as compared to NP 113. The absolute amount (mg/grain) of all the protein fractions increased during development in both NP 113 and its mutant Notch-2. During the grain development the prolamine content was substantially lower in the mutant than in the parent NP 113. The albumin + globulin content per endosperm was in general also higher in NP 113 than Notch-2. Amino acid analysis of the protein fractions did not reveal significant changes in lysine between NP 113 and Notch-2. Thus, the improvement in lysine in the mutant is primarily due to reduced synthesis of the prolamine fraction and not due to an increase in lysine in the mutant hordein fraction. Part of the improvemenht in lysine may also be due to increase in the percentage of albumin + globulin fractions which is lysine rich.Part of the Ph.D thesis submitted by S. Joshi to P.G. School, IARI, New Delhi.     

Chemical composition and nutritional potential of the tribal pulse (Bauhinia malabarica Roxb)

Proximate and mineral composition, seed protein fractions, amino acid composition, fatty acid profiles and antinutritional factors were analysed for the seeds of the tribal pulse,Bauhinia malabarica. The seeds contained higher amount of crude lipid when compared with most of the domesticated pulses. The seeds were rich in minerals like Ca, Mg and Fe. Glutelins (45%) constituted the major seed protein fraction followed by globulins which accounted for about 34%. Cystine and methionine were found to be the limiting essential amino acids; whereas tyrosine and phenylalanine content were fairly high when compared with WHO requirement pattern. The unsaturated fatty acids, oleic acid and linoleic acid, were predominant in seed lipids. Presence of antinutritional substances like total free phenols, tannins, L-DOPA and haemagglutinating activity also were analysed/assayed.     

Lack of 26 kDa globulin accompanies increased free amino acid content in rice (Oryza sativa L.) grains

Seed nitrogen content, protein composition and free amino acid content were determined in rice mutant lines with altered storage protein composition and their parental cultivars. The Lgc1 gene, which causes low glutelin content and high content of 13 kDa prolamin and 26 kDa globulin, did not affect grain weight, nitrogen content, or free amino acid content. The glb1 gene, which lacks the 26 kDa globulin, did not affect grain weight or nitrogen content, but in mutant lines with glb1 gene the content of the major free amino acids was significantly (1.3–1.5 times) higher than those of their parental cultivars. These results suggest that absence of 26 kDa globulin is accompanied by an accumulation of a high level of free amino acids in rice grains.     

Genetic control of phaseolin protein expression in seeds of common bean,Phaseolus vulgaris L.

Phaseolin, the major globulin seed storage protein of common bean,Phaseolus vulgaris L., accounts for up to 50% of the total seed protein. The rapid accumulation of phaseolin in the maturing seeds begins about 14 days after flowering and continues for some 12–14 days longer. However, the amount and rate of phaseolin accumulation, related to variation in onset, length, termination, and rate of synthesis, have been shown to vary between genotypes.Only three phaseolin electrophoretic types, designated T, S, and C after the cultivars Tendergreen, Sanilac, and Contender, respectively, have been identified among over 100 cultivated accessions. The narrow ranges of molecular weights and isoelectric points of the 14 protein polypeptides of phaseolin, as well as the homology observed from peptide mapping, suggest that the phaseolin polypeptides are similar proteins. Based on the results of crosses among cultivars having the three electrophoretic patterns, the genes controlling the polypeptides of each of the phaseolin types appear to be tightly linked, inherited in a block and the alleles are codominant.Substantial variation in phaseolin content, based on estimations using rocket immunoelectrophoresis, has been found among bean lines. Although most segregating populations show continuous distributions and quantitative inheritance, some inbred backcross lines having enhanced phaseolin accumulation appear to carry a few genes with major effects. A single gene that reduces the amount of phaseolin to less than one-half of the normal levels has been identified recently in an accession of wildP. vulgaris.     

The nutritional value of high-lysine barley genotypes

The protein quality of some improved high-lysine barley genotypes was measured chemically — including amino acid analyses and sequential protein extraction — and biologically in N-balance experiments with growing rats. The increased content of lysine, mainly due to an enhancement of the glutelin protein fraction, and other essential amino acids improved the nutritional quality as the biological value of the protein was significantly higher in the high-lysine material compared to normal. This improvement is undoubtedly of importance to meet the essential amino acid requirement of man as well as monogastric animals.     

大豆种子贮存蛋白组成及其相关分析

1988年选用14个,1989年选用15个蛋白质含量差异较大的主要为江淮地区的大豆品种,进行随机区组试验。大豆种子中球蛋白、清蛋白、醇溶谷蛋白以及谷蛋白的品种间平均分别为28.56％、7.50％、1.81％和5.71％,品种间变幅分别为6.08％、10.25％、1.03％和6.51％。清蛋白含量与全蛋白含量。谷蛋白含量与全蛋白含量之间显著正相关,清蛋白含量对种子全蛋白含量品种间差异的相对贡献最大,而球蛋白含量对品种间全蛋白含量差异的相对贡献最小。球蛋白含量与株高、分枝数和单株粒重存在显著的正相关;但株高和清蛋白含量、单株荚数及单株粒数和谷蛋白含量有显著负相关。     

Effects of glutelin and globulin on the physicochemical properties of rice starch and flour

The effect of two rice endosperm proteins, glutelin and globulin, on the physicochemical properties of rice starch and flour was investigated. Albumin, globulin, prolamin and glutelin were sequentially extracted from defatted rice flour with de-ionised water, 1.5 M NaCl, propan-2-ol and 0.1 M NaOH, respectively, followed by dialysis and lyophilisation. Globulin and glutelin were then added to pure rice starch at various concentrations, separately and together, and the pasting and textural properties of mixtures were analysed by the Rapid Visco Analyser (RVA) and TA-XT2 textural analyser, respectively. The presence of glutelin in rice starch caused an increase in pasting temperature but a decrease in the viscosity parameters of the starch paste. The concentration of glutelin was also positively correlated with the hardness and adhesive properties of the starch gel. The presence of globulin, on the other hand, resulted in a decrease in all the pasting and textural parameters except gel hardness and the changes were linearly correlated with the concentration of the protein for most of the physical parameters. When the two proteins were added to rice starch together, the outcomes in pasting and textural properties were generally dependent upon the relative concentrations of the two proteins, but were also influenced by the presence of the other two protein fractions, albumin and prolamin. The presence of globulin initially accelerated the rate of water absorption by starch during cooking while the presence of glutelin slowed it down, but in both cases, the ultimate amount of water absorbed was significantly lower than that by pure starch. The contrasting effects of the different protein fractions mean that it might be possible to manipulate the textural properties of rice starch and flour to achieve desirable sensory outcomes by varying the proportions of the protein fractions in product formulations.     

Chemical composition of the underutilized legume Cassia hirsuta L.

Seven accessions of the underutilized legume, Cassia hirsuta L., seeds collected from seven different agroclimatic regions of Tamil Nadu, India, were analyzed for proximate composition, total proteins, protein fractions, mineral profiles and selected antinutritional factors. Crude protein ranged from 15.52 to 20.74%, crude lipid 3.77–7.04%, crude fiber4.68–6.92%, ash 3.98–6.42% and carbohydrates 62.45–70.16%. Energyvalues of the seeds were 1549–1634 kJ/100 g (DM), which are comparable to those of other legumes. Data on seed protein fractions revealed thatglobulins constituted the bulk of the seed protein as in most legumes. Mineralcontents of the seeds showed greater variation. Potassium was the mostabundant mineral (1029–1786 mg/100 g), whereas manganese was low (2.1–2.2 mg/100 g). Antinutritional factors such as total free phenolics,tannins, L-DOPA and lectins were analyzed. The results of the studydemonstrated that the accessions of C. hirsuta seeds collectedfrom Tamil Nadu, India, could be good sources of some important nutrientsfor humans.