Selenium accumulation in protein fractions during germination of Se-enriched brown rice and molecular weights distribution of Se-containing proteins

This study investigated the accumulation of selenium (Se) in protein fractions of albumin, globulin, prolamin and glutelin extracted from Se-enriched brown rice and the molecular weight distribution of Se-containing proteins. Results showed that the amount of total Se (T-Se) and protein-bound Se (PB-Se) in brown rice was significantly (P < 0.05) increased after germination with 10–60 μmol/l sodium selenite. Except prolamin, the amount of all the other three protein fractions decreased significantly (P < 0.05) with the increase of germination time. Low Se concentrations had promoting effects on degradation of albumin and globulin, while no significant effects were observed on prolamin and glutelin. The accumulation of T-Se and PB-Se were in the order of albumin > glutelin > globulin > prolamin. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE) analysis showed that Se was distributed in all the proteins of which molecular weights varied from 13.6 to 121.4 kDa; however, 84.34% of Se was observed in the proteins whose molecular weights less than 36.3 kDa.     

Effect of heat,rutin and disulfide bond reduction on in vitro pepsin digestibility of Chinese tartary buckwheat protein fractions

Protein fractions (albumin, globulin, prolamin and glutelin) were extracted from defatted tartary buckwheat flour. The in vitro pepsin digestibilities of the four protein fractions were different, and albumin was more susceptible to pepsin hydrolysis. The native structure of the four protein fractions may be destroyed by heat treatment, and the digestibilities were all improved significantly (P < 0.05). Adding rutin to the digestion mixture of the four fractions did not cause a decrease in pepsin digestibility, although it did cause a significant increase in certain instances (P < 0.05). Treatment with β-mercaptoethanol (2-ME) only caused a higher initial proteolysis rate and did not increase the final digestibility distinctly except for prolamin. After pepsin digestion, the remaining proteins of unhydrolyzed albumin, globulin, prolamin and glutelin (untreated) shared some similarities. They also exhibited a minor band at 20,000 Da and a broad band at 10,000–14,000 Da.     

Physicochemical and functional properties of the protein isolate and major fractions prepared from Akebia trifoliata var. australis seed

The physicochemical and functional properties of protein isolate (API) and major protein fractions prepared from Akebia trifoliata var. australis seed were investigated. The seed contained 38.83% of oil and 17.23% of protein. Albumin (51.65%) and glutelin (46.40%) were the predominant fractions in the protein component of the seed. The major amino acids were found to be glutamic acid and aspartic acid, while the contents of sulphur-containing amino acids and threonine were very low. One to eight distinct bands with molecular weight (MW) ranging from 12.0 to 50.0 kDa were displayed by SDS–PAGE. The solubilities of API, albumin and glutelin from seeds of the A. trifoliata var. australis were the lowest at pH 4.0–5.0. The high surface hydrophobicity indices of these three proteins were observed at pH 7.0, while the excellent emulsifying properties were displayed at pH 9.0. Circular dichroism measurements indicated that API, albumin and glutelin were rich in β-strand and random coil structures.     

米胚蛋白组分的提取及功能性质研究

采用Osborne的方法,从米胚中提取清蛋白、球蛋白、醇溶蛋白和谷蛋白,4种蛋白中谷蛋白的含量最高,占72.69%,其余3种蛋白的含量相差不大;考察4种蛋白的溶解性得出,在pH5下4种蛋白的溶解性最低,随着pH的增大或减小,溶解性逐渐增加,且清蛋白和球蛋白的溶解性较醇溶蛋白和谷蛋白高;在pH11、0.1mol/L NaCl浓度、1mg/mL蛋白浓度下,加入3mL大豆油,4种蛋白的乳化活性及乳化稳定性均达到最大值。     

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     

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。     

Extractability and Chromatographic Characterization of Wheat (Triticum aestivum L.) Bran Protein

About 70% of the protein for human consumption is derived from plants, with cereals as the most important source. Wheat bran protein has a more balanced amino acid profile than that of flour. We here for the first time report the amino acid, size exclusion, and SDS‐PAGE profiles of bran Osborne protein fractions (OPFs). Moreover, we also investigated how OPFs are affected when physical barriers which entrap proteins in bran tissues are removed. Albumin/globulin is the most abundant OPF. It is richer in lysine and asparagine/aspartic acid than other OPF. Most bran albumin/globulin proteins have a molecular weight (MW) lower than 30 k and their chromatographic profiles differ from those of flour. The prolamin has high levels of proline and glutamine/glutamic acid. It is rich in proteins with a MW of 30 to 45 k and about 66 k reflecting contamination with gliadin from endosperm. The glutelin has high levels of glycine, proline, and glutamine/glutamic acid. Its protein is of intermediate and high MW with little protein with MW lower than 30 k. The high (MWs from 80 to 120 k) and low (MW around 45 k) MW glutenin subunits of flour are also present in bran. The glutelin of wheat endosperm is named glutenin. Ball milling releases albumin/globulin and glutelin but not prolamin. Not all glutelin was endosperm glutenin as a substantial part was entrapped in the aleurone cells.     

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.     

Preparation and characterisation of protein concentrates from defatted kenaf seed

Two kenaf varieties QP3 and V36 were used to obtain protein concentrates. Proximate analysis, foaming, water and oil absorption properties were studied. Significant (P < 0.05) differences were observed among the two varieties only in their content in oil and carbohydrates. The protein concentrate yield was 13.04% and 10.56%, respectively. The two varieties showed significantly different (P < 0.05) water and oil absorption capacities. QP3 showed higher foaming capacity than did V38, and it was increased with increasing salt and sugar concentration. Albumin was the main fraction representing 59.6% and 66.1% in QP3 and V36 varieties, respectively, followed by globulin, which represented 22.6% and 19.1%, respectively. The ratios of albumin, globulin, glutelin and prolamin were significantly different. Based on the data obtained from sodium dodecyl sulphate polyacrylamide gel electrophoresis, the main kenaf seed proteins present in the concentrates were five proteins with molecular weights ranging from 10 to 66 kDa. From differential scanning calorimetry data, QP3 and V36 protein concentrates had similar denaturation temperatures (82.6 and 81.8 °C, respectively).     

Amino acid profiles and quality from lotus seed proteins

BACKGROUND: Protein composition, amino acid profile and nutritional value of the lotus seed and its Osborne fractions were investigated. The seed was rich in protein with 19.85%, and showed well balanced amino acid composition compared with FAO/WHO pattern, Its nutritive properties were similar to those observed in the reference soybean protein. Phenylalanine, tyrosine, leucine and lysine were the limiting amino acids in the seed proteins. The albumin and globulin were the main protein fraction, the amino acid profile and nutritional value were close to the seed protein. RESULTS: Changes in transition temperature and thermal stability were observed through different solvent extractions. Albumin possessed the predominant thermal stability (81.4 °C) followed by globulin (74.49 °C), prolamin (69 °C) and glutelin (65.6 °C). So, solvent compositions influence the profile of AAs and their nutritive value, and aqueous solvent with 0.1 mol L^?1 NaCl was an efficient protein solubiliser. CONCLUSION: The results indicated that the extraction processes influenced the lotus seed protein quality and thermal stability. Overall, the study revealed that the lotus seed protein was nutritionally well‐balanced protein and might be of significant importance in the formulation of diets for humans. © 2012 Society of Chemical Industry     

Biologically active peptides obtained by enzymatic hydrolysis of Adzuki bean seeds

This study investigated the antioxidant and antihypertensive activities of peptides obtained from protein fractions of Adzuki bean seeds. Peptides were obtained by the use of hydrolytic enzymes in vitro under gastrointestinal conditions. A determination was made of the activity of the peptide inhibitors of the angiotensin I converting enzyme (ACE), and the antiradical and ion chelating activity of peptides from different protein fractions. The highest peptide levels after the absorption process (<7 kDa) were noted in the albumin fraction (50.69 μg/ml). Furthermore, it was found that peptides from the prolamin fraction were characterised by the highest antiradical activity and ACE inhibitory activity (IC₅₀ = 0.17 mg/ml). Peptides obtained from the globulin fraction showed the highest ability to chelate iron ions, and peptides from the glutelin fraction were characterised as being the most effective in the chelation of copper ions.     

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     

Characterization of protein fractions from chickpea (Cicer arietinum L.) and oat (Avena sativa L.) seeds using proteomic techniques

Albumin, globulin and glutelin fractions were prepared from chickpea and oat seeds using sequential extractions. Molecular characteristics of individual protein fractions were investigated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in combination with proteomic techniques. SDS-PAGE results revealed that chickpea albumin and globulin fractions (C-Ab and C-Gb) showed protein bands with molecular weights (MWs) related to subunits of legumin (11S globulin) and pea vicilin (7S globulin); oat protein fractions (O-Ab, O-Gb and O-Gt) showed most protein bands with MWs related to subunits of oat 12S globulin (avenalin). With proteomic analysis, eighteen tryptic peptides from chickpea globulin fraction showed sequence homology that corresponded to chickpea legumin α- and β-subunit (NCBI accession number: gi|6273402; theoretical mass 56,216 Da) while sixteen tryptic peptides from chickpea albumin fraction (C-Ab) were identified as chickpea provicilin precursor (NCBI accession number: gi|82173888; theoretical mass 51,390 Da); fifteen tryptic peptides from oat protein fractions were identified with origin from oat 12S seed storage globulin 1 (NCBI accession number: gi|134918; theoretical mass 58,508 Da). The identified tryptic peptide, ALIVPQNFAIAAK, was commonly found in chickpea glutelin fraction (C-Gt), rice glutelin fraction (R-Gt), and oat albumin, globulin and glutelin fractions (O-Ab, O-Gb and O-Gt).     

Angiotensin I-converting enzyme-inhibitory peptide fractions from albumin 1 and globulin as obtained of amaranth grain

A number of biopeptides promoting health benefits have been isolated from food-protein hydrolysates and can be released during enzymatic digestion. Antihypertensive peptides can be part of protein fractions from amaranth grain. The objective of this work was to obtain ACE-inhibitory peptide fractions from albumin 1 and the globulin of amaranth (Amaranthus hypochondriacus) grain. Albumin 1 and globulin were hydrolysed with alcalase; hydrolysis was monitored by proteolytic degradation and by ACE-inhibitory activity. The highest ACE-inhibitory activity was 40% and 35% as obtained after 18 and 15 h hydrolysis for albumin 1 and globulin, respectively. Further separation and purification of the ACE-inhibitory peptide fractions were carried out by gel filtration and C₁₈ RP–HPLC. The IC₅₀ was 0.35 ± 0.02 mg/ml for albumin 1 peptide fraction and 0.15 ± 0.03 mg/ml for globulin peptide fraction. Albumin 1 peptide fraction showed an competitive mode of ACE inhibition, whereas the globulin peptide fraction was competitive. The globulin peptide fraction may have one of the most active naturally-occurring ACE-inhibitory peptides.     

4种花生粕蛋白的理化性质及功能特性研究

以花生粕为原料,采用分级提取工艺提取花生清蛋白、球蛋白、醇溶蛋白和谷蛋白,研究4种花生粕蛋白的理化性质和功能特性。扫描电镜观察,4种花生粕蛋白的形态结构各不相同。SDS-PAGE法测定分子量表明,清蛋白含有4种亚基,相对分子量分别为70kDa、40kDa、30kDa、25kDa和15kDa;醇溶蛋白含有2种亚基,分子量分别为25kDa和15kDa;球蛋白含有5种亚基,分子量分别为40kDa、38kDa、30kDa、25kDa和15kDa;谷蛋白含有4种亚基,分子量分别为40kDa、30kDa、25kDa和15kDa。花生清蛋白、醇溶蛋白、球蛋白、谷蛋白的等电点分别为pH3.6、pH5.2、pH4.6、pH5.0。功能性质研究表明,球蛋白的持水性最好,为1.52mL/g,其次为谷蛋白1.10mL/g,清蛋白和醇溶蛋白的持水性较低分别为0.49mL/g、0.14mL/g;清蛋白的持油量相对较高为8.21mL/g,其次为球蛋白为7.16mL/g,谷蛋白和醇溶蛋白的持油量相对较低,分别为3.82mL/g和5.49mL/g;清蛋白的乳化性和乳化稳定性相对较高,乳化能力EC值和乳化稳定性ES值分别为71.4% 和83.33%,谷蛋白次之,EC和ES值分别为66.7% 和82.86%,醇溶蛋白和球蛋白相对较低,EC值分别为64.0% 和62.2%,ES值分别为82.35% 和76.67%。综上,花生粕清蛋白的持油性、乳化性和乳化稳定性相对较好。     

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.     

IN VITRO DIGESTIBILITY OF CHINESE TARTARY BUCKWHEAT PROTEIN FRACTIONS: THE MICROSTRUCTURE AND MOLECULAR WEIGHT DISTRIBUTION OF THEIR HYDROLYSATES

Our previous study showed that in vitro pepsin digestibility of Chinese tartary buckwheat protein was relatively low compared to those of other edible seeds. In vitro pepsin digestibilities of four protein fractions of tartary buckwheat, microstructure and molecular weight (MW) distributions of hydrolysates were investigated. In vitro pepsin digestion assay showed that the digestibilities of tartary buckwheat protein fractions were albumin (81.20%), globulin (79.56%), prolamin (66.99%) and glutelin (58.09%). Scanning electron microscopy showed that albumin and globulin fractions were digested by pitting from the outer surface to the inner part and were more digestible, while prolamin and glutelin fractions resisted digestion because only the outer surfaces of their protein bodies were digested and the interior was protected. MW distribution of the hydrolysates from the four protein fractions was determined by high‐performance liquid chromatography. The hydrolysates of albumin mainly consisted of polypeptides with lower MW. The hydrolysates of glutelin had larger polypeptides together with small and medium‐sized peptide fractions.     

Effects of jugular-infused lysine, methionine, and branched-chain amino acids on milk protein synthesis in high-producing dairy cows

In addition to lysine and methionine, current ration-balancing programs suggest that branched-chain amino acid (BCAA) supply may also be limiting in dairy cows. The objective of this study was to investigate whether BCAA, leucine, isoleucine, and valine become limiting for milk protein synthesis when methionine and lysine supply were not limiting. Nine multiparous Holstein cows with an average milk production of 53.5 ± 7.1 kg/d were randomly assigned to 7-d continuous jugular infusions of saline (CTL), methionine and lysine (ML; 12 g and 21 g/d, respectively), or ML plus leucine, isoleucine, and valine (ML+BCAA; 35 g, 15 g, and 15 g/d, respectively) in a 3 × 3 Latin square design with 3 infusion periods separated by 7-d noninfusion periods. The basal diet consisted of 40% corn silage, 14% alfalfa hay, and a concentrate mix, and respectively supplied lysine, methionine, isoleucine, leucine, and valine as 6.1, 1.8, 4.7, 8.9, and 5.3% of metabolizable protein. Dry matter intake (23.9 kg/d), milk yield (52.8 kg/d), fat content (2.55%), fat yield (1.33 kg/d), lactose content (4.77%), lactose yield (2.51 kg/d), and milk protein efficiency (0.38) were similar across treatments. Protein yield and protein content were not significantly different between ML (1.52 kg/d and 2.88%, respectively) and ML+BCAA (1.51 kg/d and 2.83%, respectively), but they were significantly greater than that of CTL (1.39 kg/d and 2.71%). Cows that received ML+BCAA had less milk urea nitrogen content (10.9 mg/dL) compared with milk of CTL cows (12.4 mg/dL) and ML cows (11.8 mg/dL). Whereas high-producing cows responded positively to methionine and lysine supplementation, no apparent benefits of BCAA supplementation in milk protein synthesis were found. Infusion of BCAA may have stimulated synthesis of other body proteins, probably muscle proteins, as evidenced by decreased milk urea nitrogen.     

Changes in gross chemical composition with emphasis on lipid and protein fractions during germination of fenugreek seeds

Germination of fenugreek seeds for 3 and 5 days increased moisture, crude protein, crude fibre, non-protein nitrogen and ash contents, but total lipids and carbohydrates were decreased. Marked increases of Na and P, as well as Mg and Zn were observed. Triacylglycerols decreased continuously and this was accompanied by an increase in free fatty acids, monoglycerides, sterolesters and polar lipids. Albumin was the major protein fraction followed by globulin, glutelin and prolamin. Germination increased glutelin and non-protein nitrogen; the other fractions were decreased. The protein in vitro digestibility was improved after germination. PAGE showed a marked dissociation of albumin and globulin fractions after germination.     

Purification and characterization of antioxidative peptides derived from rice bran protein hydrolysates

Rice bran protein fraction (RBPF)—albumin, globulin, glutelin and prolamin were hydrolyzed with proteases M, N, P, S and pepsin under their optimal conditions for 24 h. Hydrolysates of various hydrolysis periods were collected and subjected to peptide mapping and the antioxidative activity measured by the 2,2-Azino-bis-3-ethylbenzothiazoline-6-sulfonic Acid (ABTS) method. Protease M hydrolysates showed high degree of hydrolysis (DH), but low antioxidative activity. On the contrary, pepsin hydrolysates showed low DH with high activity. Albumin and globulin hydrolysates had higher DH values, but lower values for glutelin and prolamin. The globulin hydrolysate (Opep2) from 2 h-pepsin hydrolysis was separated by using three consecutive purification steps with RP-HPLC. Nineteen antioxidative peptides were isolated and their amino acid sequences were determined by a gas-phase protein sequencer and MALDI-TOF mass spectrometry. These peptides were composed of 6–30 amino acid residues with molecular masses ranging from 670–3,611 Da. Tyr-Leu-Ala-Gly-Met-Asn had the highest antioxidative activity among them.