Japanese barnyard millet (Echinochloa frumentacea): Protein content,quality and SDS–PAGE of protein fractions

Total protein from five varieties of Japanese barnyard millet (Echinochloa frumentacea,) was separated into albumin/globulin, prolamin and glutelin fractions. The protein fractions were examined by sodium dodecylsulphate polyacrylamide gel electrophoresis. Total protein of the varieties ranged from 110·5 to 139·3 mg g^?1 of which 11·3–17·2% was albumin/globulins, 6·8–9·3% prolamins, 7·5–11·6% prolamin–like, 5·9–9·1% glutelin-like and 39·3–54·4% true glutelins. Amino acid analyses of the total protein showed that the varieties had essentially the same ammo acid composition. With the exception of lysine the amino acid levels adequately matched the provisional FAO scoring pattern. The amino acid composition of the protein fractions was also very similar. Electrophoretic analysis showed that the albumin/globulin fraction contained three or four components; the prolamin and glutelin fractions each had five components. The glutelin fraction had higher molecular weight components than the other two fractions.     

Seed proteins of finger millet and their amino acid composition

Determination of the protein content and amino acid composition of finger millet samples from white-grain and brown-grain varieties, revealed wide variations in protein and amino acid composition. Solubility fractionation of proteins showed that prolamin and glutelin constitute the major protein fractions. White-grain varieties have higher prolamin and lower glutelin levels than the brown-grain varieties. The amino acid composition of three protein fractions of one white and one brown variety are reported. These studies suggest that it should be possible to provide high amounts of good quality protein in lines developed from white×brown seed crosses.     

Studies on the proteins of kodo millet (Paspalum scrobiculatum)

Studies on the proteins of five varieties of kodo millet showed that there are hardly any varietal differences. Fractionation of the proteins showed that glutelin is the major storage protein. The amino acid compositions of the protein of the five varities of kodo millet and the individual protein fractions of two varieties were determined. Lysine is the most limiting amino acid followed by methionine and cystine. There is no significant negative correlation between lysine levels and protein content. Sodium dodecylsulphate polyacrylamide gel electrophoresis patterns indicate great similarities in the different varieties.     

Studies on the amino acid and mineral content of buckwheat protein fractions

Protein fractions were extracted by successive extraction and analysis method in four buckwheat varieties including Japanese spring buckwheat, Japanese summer buckwheat, Yuqiao No. 1 and Yuqiao 6-21. The amino acid and the mineral content of each protein fraction were also analyzed in this paper. The basis for the data on the experiment was the relationship between protein fractions and amino acids, and the mineral contents of protein fractions. The distribution and proportion of amino acids and the minerals in each protein fraction in the buckwheat kernel as well as the nutritional value of buckwheat kernel were discussed. The results showed that there is a high amount of soluble protein and very low amount of prolamin in the buckwheat kernel. Leucine is the first limited amino acid in buckwheat, and buckwheat protein is only a seminutritional protein. The buckwheat kernel is rich in K, Zn in the albumin, Ca, Mg and Mn in the globulin, Na in the prolamin and glutelin.     

Relationships between amino acid composition and nitrogen of foxtail (Italian) millet (Setaria italica) grain of different varieties

The amino acid composition of 13 samples of foxtail millet (Setaria italica L) from six Chinese and one French varieties was determined as a function of their N content (N), which ranged from 1·82 to 3·65 g per 100 g of grain DM. The levels of amino acids in grain DM increased linearly with N with correlation coefficients close to 1 for most of them regardless of foxtail millet genotype or phenotype. Thus simply knowing N enables one to predict the amino acid composition of any foxtail millet grain sample. Amino acids in crude protein of grain (g 16 g^?1N)changed as quadratic functions of N, which decreased for glycine, cysteine, tysine, histidine and arginine, remained nearly constant for valine, threonine, tyrosine, methionine and aspartate plus asparagine, and increased for other amino acids. Foxtail millet appeared as the only cereal in which lysine is the only limiting essential amino acid. However, the lysine score was low and intermediate between that of maize and sorghum, falling from 48 to 31 % when N increased from 1·82 to 3·65 g per WO g DM. The N-to-protein conversion factor strongly increased with N and was the highest of all cereals within the N range studied. The results also showed that the composition of storage proteins accumulated in grains remained constant, with a prolamin to glutelin ratio close to three and independent of grain protein content.     

Walnuts (Juglans regia L): proximate composition,protein solubility,protein amino acid composition and protein in vitro digestibility

Walnuts contained 16.66% protein and 66.90% lipids on a dry weight basis. Non‐protein nitrogen values ranged from 6.24 to 8.45% of the total nitrogen when the trichloroacetic acid concentration was varied within the range 0.25–1.0 M . Albumin, globulin, prolamin and glutelin respectively accounted for 6.81, 17.57, 5.33 and 70.11% of the total walnut proteins. Walnut proteins were minimally soluble at pH 4.0. The majority of total walnut protein polypeptides had estimated molecular weights in the range 12 000–67 000. The Stokes radius of the major protein in walnuts (glutelin fraction) was 66.44 ± 1.39 Å. Lysine was the first limiting essential amino acid in total walnut proteins as well as in the globulin and glutelin fractions. Leucine and methionine plus cysteine were the second limiting essential amino acids respectively for the prolamin and albumin fractions. Hydrophobic and acidic amino acids dominated the amino acid composition in all protein fractions. Native and heat‐denatured walnut glutelins were easily hydrolysed by trypsin, chymotrypsin and pepsin in vitro. © 2000 Society of Chemical Industry     

Characterisation of storage protein from selected varieties of foxtail millet (Setaria italica (L) Beauv)

Protein content of the three selected varieties of Setaria italica (L) Beauv ranged from 91·7 to 112·0 g kg⁻¹. Fractionation of seed protein showed that prolamin is the major storage protein and the differences in protein content among them is mainly due to differences in the total prolamin accumulation. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) pattern of different protein fractions; namely, albumin/globulin, prolamins and glutelins and RP-HPLC (reversed-phase high-performance liquid chromato-graphy) of the total prolamins showed great similarity among the three selected varieties. Accumulation of total protein and protein fractions during seed maturation was studied in all the three selected varieties. Fractionation of total protein in the grain collected at 5, 10, 15, 20, 25, 36 days after anthesis revealed that prolamin is the major storage protein ( > 50% of total protein) during all the stages of seed development. The SDS-PAGE pattern of the total prolamin at different stages of seed development showed no variation among the selected varieties. Prolamin polypeptide synthesis commenced as early as 5 days after flowering (DAF) and completed before 10 DAF. Thereafter, only quantitative increase was observed. Immunoblotting of the total prolamin from S italica probed with 20 kDa prolamin protein antibody of kodo millet (Paspalum scrobiculatum) showed that the two millets are immunologically related to one another. © 1998 SCI.     

Nutritional evaluation of high protein genotypes of pearl millet (Pennisetum americanum (L.) leeke)

Three high protein (HP) inbred lines (700112, WC-190 and B-816) of pearl millet were studied for their nutritional quality and the results compared with those of normal protein varieties. The protein content of HP lines showed an average increase of 60% but the starch and soluble sugars together and the fat content decreased by 40 and 20%, respectively. Total dietary fibre of the HP lines was about 10% higher. A decrease (about 20%) in the albumin fraction was associated with an increase in prolamin in HP lines. The amino acid composition of the HP lines remained normal except for an approximately 16% decrease in lysine. However, the total amount of lysine in the sample increased by an average of 37%, as a result of a substantial increase in protein content. True protein digestibility was very high for each line but, expectedly, the biological value was markedly lower. The values for utilisable protein were highest for high protein genotypes. Digestible energy was high in all the genotypes, although slightly lower values were obtained for the HP lines; this was attributed to the fibre fractions.     

The effect of nitrogen nutrition on the lysine content and protein composition of barley seeds

Increases in the total grain nitrogen of barley (cv. Julia and Bomi), resulting from increased N fertilisation, are accompanied by decreases in the relative lysine content of the grain proteins. Extraction of the grain proteins shows that the high-N grain have increased relative and total amounts of the lysine-poor storage protein (hordein) fraction. There are only small increases in the total amounts of the other more lysine-rich protein fractions (salt-soluble proteins and glutelin+residue proteins). There is also little or no effect on the amino acid compositions of the fractions. Consequently, it is concluded that the decreased relative lysine content of the whole grain is due to the increased synthesis of hordein. Further evidence for this hypothesis comes from similar studies with the high-lysine mutant, Risø 1508, in which the high-lysine character results from a decrease in the relative and total amount of hordein. The hordein fraction does not increase disproportionately with increasing grain N, nor does the relative lysine content of the grain decrease. There is no effect of grain N on the polypeptide compositions of the salt-soluble proteins of either line. However, in Bomi and Julia increased grain N does result in an increase in the relative amount of sulphur-poor ‘C’ hordein polypeptides in the hordein fraction. It is speculated that this results from limitation of S relative to N.     

Electrophoretic Characterization and Functional Properties of Rice Proteins from Indian Rice Cultivars

Rice protein isolates and extracts of protein fractions were prepared from Indian rice cultivars, namely, Jaya, HKR-120, P-44, Sharbati, Bas-370, and HBC-19. The protein extracts were characterized using SDS-PAGE. The total protein contents of rice cultivars ranged from 5.46 to 7.02 g/100 g sample with albumin and glutelin fractions showing the highest variability among rice cultivars. The electrophoretic patterns of protein fractions exhibited many varietal differences with glutelin fraction revealing the most heterogeneous (10–17 polypeptide units) and prolamin fraction revealing the most homogenous polypeptide composition (3 polypeptide units). The alkali extracted rice endosperm protein isolates showed favorable emulsifying and foaming capacities particularly at an alkaline pH of 11. The total protein content was significantly correlated positive with foaming capacity (r = 0.917, p < 0.01) and negative with oil absorption capacity (r =??0.914, p < 0.05). The total protein content was also correlated significantly positive with cooking time (r = 0.956, p < 0.01) and cooked grain hardness (r = 0.966, p < 0.01).     

The protein and the amino acid composition of some rice and maize varieties grown in North Vietnam

The protein and amino acid composition of several rice and maize varieties grown in North Vietnam, and their digestibility, was determined. The protein content (N×5.95) of rice cultivars ranged from 7.0 to 10.8% of which 70–80% was in the glutelin fraction. The true digestibility was relatively good (87.6–91.8%). In general, lysine and threonine were found to be the first and second nutritionally limiting amino acids, except for two varieties, which had a low content of sulphur-containing amino acids. The protein content of maize cultivars ranged from 8.4 to 12.9%. Zein and glutelin were the main components occurring in near-equal quantities (except in the opaque-2 mutant). The overall amino acid distribution was similar to that of maize grown in other countries. Lysine levels were relatively low, and it was the first nutritionally limiting amino acid, except in the opaque-2 cultivar, tryptophan being the second one. The digestibility ranged from 87.5 to 91.1%.     

Fractionation and characterization of tartary buckwheat flour proteins

Protein fractions (albumin, globulin, prolamin and glutelin) were extracted from defatted tartary buckwheat flour. Albumin was the predominant protein fraction (43.8%) followed by glutelin (14.6%), prolamin (10.5%), and globulin (7.82%). Albumin was relatively rich in histidine, threonine, valine, phenylalanine, isoleucine, leucine and lysine. Globulin had high levels of methionine and lysine. Prolamin was high in histidine, threonine, valine, isoleucine, and leucine. Glutelin was rich in histidine, threonine, valine, isoleucine, and leucine. SDS–PAGE analysis, under non-reductive and reductive conditions, showed that disulfide bonds existed in the four fractions. Non-reduced albumin showed major bands at 64, 57, 41, and 38 kDa, and globulin at 57, 28, 23, 19 and 15 kDa. Reduced albumin and globulin shared two common bands at 41 and 38 kDa. Reduced prolamin showed major bands at 29, 26, 17 and 15 kDa. In vitro pepsin digestibility of the four fractions (from high to low) was: albumin > globulin > prolamin and glutelin.     

Val bean (Lablab purpureus L.) proteins: composition and biochemical properties

Val bean (Lablab purpureus L.) proteins were fractionated using the Osborne protein fractionation scheme and biochemically characterized. The seed flour contained 302 g kg^?1 protein (micro‐Kjeldahl N × 6.25) on a dry weight basis. Albumin, globulin, prolamin, and glutelin accounted for 22.8%, 45.1%, 1.8% and 30.3%, respectively, of the total soluble seed proteins. Among the solvents tested, 0.1 mol L^?1 aqueous NaOH was the most effective protein solubilizer. Isoelectric focusing indicated the seed proteins to be predominantly acidic (pI range was ～4–7). Val globulin is a glycoprotein composed of at least three polypeptides in the molecular mass range 51–64 kDa. Albumin fraction had the highest trypsin inhibitory activity, while the globulin fraction registered the highest hemagglutinating activity. Sulfur amino acids were the first limiting amino acids in the total seed proteins. The proportion of essential to total [E/T(%)] amino acids for the bean flour was 36.97%. Among the protein fractions, glutelin fraction had the highest E/T (42.86%) followed by albumin (41.57%), globulin (39.87%), and prolamin (39.15%). Native globulin, although resistant to pepsin, was effectively digested in vitro upon moist heat (100 °C, 30 min) denaturation. Copyright © 2007 Society of Chemical Industry     

Assessment of protein fractions of three cultivars of <Emphasis Type="Italic">Pisum sativum</Emphasis> L.: effect of germination

Protein composition, nitrogen and amino acid content of Osborne fractions in different cultivars of Pisum sativum L. (cv. ucero, cv. ramrod and cv. agra) seeds were investigated and the effect of germination on these parameters was also analysed. Albumins comprised the main protein fraction in raw seeds, globulins were constituted mainly by vicilin, with a smaller proportion of the glutelin and prolamine fractions. Regarding the amino acid profile of pea protein fractions, although differences among pea cultivars were found, in general albumin, glutelin and prolamine fractions presented Asp, Glu and Gly as the major non-essential amino acids (NEAA) and Lys as the main essential amino acid (EAA). The globulin fraction, however, presented Asp, Glu, Gly and Arg as the major NEAA and Leu, Phe, Lys, and Thr as the main EAA. In general, the albumin fraction accounted for more sulphur amino acids and Lys, followed by the glutelin + prolamine fraction. Germination caused an increase in the total protein content of P. sativum cv. ucero and P. sativum cv. ramrod. In the albumin fraction a wide number of proteins underwent degradation and convicilin disappeared from the globulin fraction of pea sprouts whilst vicilin and legumin decreased slightly. In general, all the Osborne fractions of pea sprouts presented higher EAA contents than raw seeds. The estimated essential amino acid indexes of protein fractions for P. sativum cv. ucero (EAAI_adult and EAAI_egg) improved with the germination process whilst for P. sativum cv. ramrod and P. sativum cv. agra depended on the Osborne fraction.     

苦荞蛋白质的低消化性研究Ⅰ——热处理、还原二硫键及添加芦丁对其体外消化率的影响

采用Osborne分级分离的方法从苦荞粉中制备得到清蛋白、球蛋白、醇溶蛋白和谷蛋白。体外消化率测定结果表明,四种蛋白组分的消化率均低于对照-小麦胚分离蛋白和大豆分离蛋白,并且四种组分的体外消化率也存在不同程度的差别:清蛋白最高,谷蛋白最低。热处理可以明显提高苦荞粉四种蛋白组分的体外消化率。添加芦丁不但没有降低四种蛋白组分的体外消化率,反而均有一定程度的提高。二硫键的破坏,除醇溶蛋白得以提高之外,对于其它三种组分只是提高了初始水解速度,最终体外消化率没有明显提高。体外消化实验后,四种蛋白组分所剩的残渣蛋白SDS-PAGE分析结果表明:这些残渣蛋白的谱带存在相似之处:在20kDa处有一条很窄的谱带,在14～10kDa处的谱带较宽。     

Effects of gamma irradiation on edible seed protein,amino acids and genomic DNA during sterilization

This work describes radiation-induced effects on edible seed protein profiles, carbohydrates, amino acids and genomic DNA during gamma sterilization. The total protein and carbohydrate was decreased with increasing dose compared to control samples. Oryzasativa L. Cv-2233 exhibited a minimum effect in terms of its loss in total soluble protein content, compared to other seeds at 6 kGy, and the soluble protein fraction, containing 14–16 kDa albumins and 22 kDa globulin, was unchanged up to 6 kGy. In Cicer arietinum, the effect of gamma rays was more pronounced on albumin and prolamin with respect to glutelin and globulin. The easy-to-digest and difficult-to-digest proteins were not significantly affected up to 4 kGy. However, the soluble free amino acids of all the seeds increased with increasing dose. The total DNA content and band intensities both decreased with increasing absorbed dose; however, the band positions were unchanged for all seed types.     

Protein classification of beach pea (Lathyrus maritimus L.)

Protein fractionation of whole seed, cotyledons and hulls of beach pea (Lathyrus maritimus L.) was carried out. Surface topography of pea protein isolates and protein fractions, as well as their polyacrylamide gel electrophoresis (PAGE), was studied. The nitrogen solubility of beach pea seed meal was minimum at pH 4.5. Globulin was the major protein fraction present and its content in whole seeds (57%), cotyledons (62%) and hulls (24%) of beach pea was lower than those of other common pea cultivars; the same was true for its albumin content. However, glutelin content in beach pea seed and its parts (cotyledons and hulls) was higher than those of other pea cultivars. The albumin fraction contained the highest amount of total sulphur-containing amino acids followed by glutelin, globulin and prolamine. The amount of sulphur-containing amino acids in beach pea protein fractions was higher than in other peas. Predicted biological value of albumin and glutelin fractions of beach pea was also higher than green pea and grass pea. Beach pea seed protein fractions showed different UV spectra from other pea cultivars. Protein isolate and protein fractions of beach pea seeds showed similar topographical characteristics to those of other peas. Major polypeptide bands in the range of 35–47 kDa in the protein isolate, as well as protein fractions for beach pea, were detected by PAGE.     

Isolation,fractionation, and some chemical studies on Kleinhovia hospita Linn. seed protein

Protein was extracted from the seeds of Kleinhovia hospita Linn., which is being a nonconventional source. Extraction of K. hospita seed protein at various pH values in aqueous solution and at pH 7, different salt concentrations were done. Fractionation of protein from seeds was performed to separate albumin, globulin, prolamin, and glutelin. The amino acid compositions of total protein isolate (TPI) and the fractions were determined. A total of 15 amino acids were identified including 9 essential amino acids. Gel filtration by Sephadex G-100 revealed the presence of three components in the TPI. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis of TPI and fractions showed different polypeptide bands having molecular weights ranging from 12 to 42 kDa approximately. Scanning electron microscopic study of TPI and fractions revealed the surface topology of the protein.     

发芽对糙米蛋白质及氨基酸组成特性的影响

研究了发芽过程中糙米蛋白质和氨基酸含量和组成特性的变化,并与精米和糙米比较,评价了发芽糙米蛋白质的营养价值。结果表明,发芽可以增加糙米蛋白质、总氨基酸和必需氨基酸的含量,必需氨基酸指数提高。发芽过程中清蛋白和谷蛋白含量均呈上升趋势,球蛋白含量呈下降的趋势。除发芽24h外,醇溶蛋白的含量在整个发芽期间也呈现增加趋势,增加的蛋白质主要是谷蛋白。各发芽阶段糙米的必需氨基酸和总氨基酸的比值均大于55%。18种氨基酸中有12种氨基酸的含量在发芽24h最高。发芽后赖氨酸和苏氨酸的含量增加较明显,但仍是限制性氨基酸;丝氨酸、谷氨酸、精氨酸和组氨酸含量均有所增加。发芽24h时EAAI最大,与FAO/WHO模式相比,EAA组成模式更加合理。发芽可以提高糙米蛋白质的营养价值,改善籼稻米的品质,其中以发芽24h为宜。     

Preparation and characterization of high‐quality rice bran proteins

Rice bran contains 120–200 g kg^?1 protein in addition to a large amount of fat, carbohydrate, and phytic acid. Rice bran protein (RBP) fractions were refined by a two‐step preparation to eliminate residual carbohydrate. The first step involved the sequential extraction of defatted rice bran into RBP fractions using their distinct solubility to give 37 g kg^?1 of albumin, 31 g kg^?1 of globulin, 27 g kg^?1 of glutelin, and 2 g kg^?1 of prolamin. In the second step, carried out by dissolving in respective solvent and isoelectric precipitation, the protein content of each fraction increased from 69% to 97% for albumin, from 71% to 90% for globulin, from 74% to 83% for glutelin, and from 18% to 20% for prolamin. The low protein content in the prolamin fraction might be due to its low solubility in the protein assay. Emulsifying stability index and surface hydrophobicity increased in the second‐step preparation of albumin and globulin, but not of glutelin. Emulsifying properties of RBPs were lower than that of a soybean protein isolate. Denaturation temperatures and enthalpy values of denaturation for albumin, globulin, glutelin, and prolamin were 50.1 °C/1.2 J g^?1, 79.0 °C/1.8 J g^?1, 74.5 °C/3.0 J g^?1, and 78.5 °C/8.1 J g^?1, respectively. No significant differences in the denaturation temperatures and enthalpy values of denaturation of RBP fractions were obtained with these two‐step preparations (P < 0.05). Copyright © 2007 Society of Chemical Industry