Characterization of yam bean (Pachyrhizus erosus) proteins

Seed proteins from Mexican yam bean seeds (Pachyrhizus erosus L.) were sequentially extracted according to the Osborne classification. Albumins were the major fraction (52.1-31.0%), followed by globulins (30.7-27.5%). The minor protein fraction was prolamins (0.8%). Defatting with chloroform/methanol remarkably affected the distribution of protein solubility classes; albumins were the most affected fraction (4.3-17.5%). Electrophoretic patterns of albumins showed bands at 55, 40, 35, and 31 kDa. After reduction of the globulin fraction exhibited two triplets, one from 35 to 31 kDa and the second from 19 to 21 kDa, these could be compared to the acid and basic polypeptides of 11S-like proteins. Prolamins showed one band at 31 kDa, and glutelins after reduction showed three main bands at 52, 27, and 14 kDa. Trypsin inhibitors were assayed in saline extracts; the values found (1232-2608 IU/g of meal) were lower than those of other legumes. In general, yam bean seed proteins showed an excellent balance of all essential amino acids; albumins contain the highest amount of essential amino acids.     

Biochemical Characterization and Quantification of the Storage Protein (Secalin) Types in Rye Flour

Kernels of the rye cultivars Danko and Halo were milled into white flour and compared with flour of the wheat cultivar Rektor. Flour proteins were extracted stepwise with a salt solution (albumins‐globulins), 60% ethanol (prolamins), and 50% 2‐propanol under reducing conditions (glutelins). The quantification by reversed‐phase HPLC indicated that the extractable proteins of both rye flours consisted of ≈26% albumins‐globulins, 65% prolamins, and 9% glutelins. Compared with wheat flour, rye flours comprised significantly higher proportions of nonstorage proteins (albumins‐globulins) and lower proportions of polymerized storage proteins (glutelins). SDS‐PAGE revealed that the prolamin fractions of rye contained all four storage protein types (HMW, γ‐75k, ω, and γ‐40k secalins), whereas the glutelin fractions contained only HMW and γ‐75k secalins. The quantification of secalin types by RP‐HPLC showed a close relationship between the two cultivars.The γ‐75k secalins contributed nearly half (≈46%) of the total storage proteins, followed by γ‐40k secalins (24%) and ω secalins (17%); HMW secalins (≈7%) were minor components, and 6% of eluted proteins were not identified. The amino acid composition of γ‐40k secalins corresponded to those of γ‐gliadins of wheat, whereas γ‐75k secalins were characterized by higher contents of glutamine and proline. Matrix‐assisted laser desorption/ionization and time of flight mass spectrometry (MALDI‐TOF MS) indicated molecular masses of about 52,000 (γ‐75k) and 32,000 (γ‐40k), respectively. N‐terminal amino acid sequences were homologous with those of wheat γ‐ gliadins except for position 5 (asparagine in γ‐75k and glutamine in γ‐40k secalins) and position 12 (cysteine in γ‐75k secalins). The N‐terminal amino acid sequences of HMW and ω‐secalins were homologous with those of the corresponding protein types of wheat. Gel‐permeation HPLC of prolamin fractions revealed that rye flours contained a significantly higher proportion of ethanol‐soluble oligomeric proteins than wheat flour.     

Jojoba seed meal proteins associated with proteolytic and protease inhibitory activities

The jojoba, Simmondsia chinensis, is a characteristic desert plant native to the Sonoran desert. The jojoba meal after oil extraction is rich in protein. The major jojoba proteins were albumins (79%) and globulins (21%), which have similar amino acid compositions and also showed a labile thrombin-inhibitory activity. SDS-PAGE showed two major proteins at 50 kDa and 25 kDa both in the albumins and in the globulins. The 25 kDa protein has trypsin- and chymotrypsin-inhibitory activities. In vitro digestibility of the globulins and albumins resembled that of casein and soybean protein concentrates and was increased after heat treatment. The increased digestibility achieved by boiling may be attributed to inactivation of the protease inhibitors and denaturation of proteins.     

Molecular cloning of 11S globulin and 2S albumin, the two major seed storage proteins in sesame

Insoluble 11S globulin and soluble 2S albumin, conventionally termed alpha-globulin and beta-globulin, are the two major storage proteins and constitute 80-90% of total seed proteins in sesame. Two full-length cDNA clones were sequenced and deduced to encode sesame 11S globulin and 2S albumin precursors, respectively. Deduced amino acid composition reveals that 2S albumin, but not 11S globulin, is a sulfur-rich protein. Three abundant polypeptides of 50-60 kDa were resolved on SDS-PAGE when seed-purified 11S globulin was prepared in nonreducing conditions. Immunological analysis suggests that these three polypeptides are encoded by homologous genes. Immunodetection on the overexpressed protein of the 11S globulin clone in Escherichia coli indicates that this clone encodes the precursor protein of one of the three purified 11S globulin polypeptides.     

Compared use of HPLC and FZCE for cluster analysis of Triticum spp and for the identification of T. durum adulteration

Wheat quality criteria continually evolve in response to market pressure and consumer preference. Characterization of cereal cultivars for quality and agronomic properties, have widely shown the importance of the protein content to ensure good quality products. The aim of this work is a comparison of reversed-phase high performance liquid chromatography (RP-HPLC) and free zone capillary electrophoresis (FZCE) in the identification of Italian wheat cultivars and detection of durum wheat flour adulteration. Mainly alcohol soluble (gliadins) and water soluble (albumins) proteins were extracted from 14 common wheat cultivars and from 9 durum wheat cultivars. In RP-HPLC chromatograms, wheat albumins and gliadins eluted between 3 and 9 min and between 10 and 42 min, respectively. Even if the chosen chromatographic conditions (reversed phase) did not permit a complete resolution of hydrophilic proteins such as albumins, a good reproducibility was observed for both albumins and gliadins. In FZCE electropherograms, wheat albumins and gliadins migrated between 8 and 14 min and 16-25 min, respectively. A good reproducibility was found for wheat albumins, while the relatively poor reproducibility of gliadin fractions was a consequence of the selected separation conditions aimed to separate in the same run either hydrophilic (albumins) and alcohol-soluble (gliadins) proteins. The principal component analysis (PCA) of HPLC and FZCE data evidenced that both techniques allowed the univocal identification of the great proportion of investigated wheat cultivars. Three peaks were exclusively detected in RP-HPLC chromatograms of common wheat cultivars, while three unique peaks were found in FZCE electropherograms of common wheat cultivars. These peaks were investigated as a basis for detecting and estimating the adulteration of durum wheat flour with flour from common wheat. The direct relationship between the area of the peaks and adulteration level enabled standard curves to be constructed. The standard curves showed that adulteration may be quantified by either RP-HPLC or FZCE.     

Surface physicochemical properties of globulin-P amaranth protein

Globulin-P, the polymerized 11S amaranth globulin, is composed of 280 kDa unitary molecules (UM, 23%) and aggregates larger than 500 kDa (A, 70%). Antibodies against these proteins were prepared to study their surface characteristics and to assess their homology with other storage proteins. Results showed that globulin-P unitary molecules and aggregates had similar reactive surfaces. A polypeptide of 56 kDa was found to be the most reactive to the antibodies assayed, followed by the acidic polypeptides. Such results support previous information, according to which these polypeptides appeared to be the most exposed on the molecule surface. Globulin-P fraction presented cross-reactivity with the remaining amaranth protein fractions: 11S-globulin, glutelins, and albumins. Globulin-P and 11S-globulin showed similar reactive surfaces whereas glutelin and albumins presented a lower cross-reactivity. The reactivity of the glutelin fraction depended on its sequence. Globulin-P fraction presented cross-reactivity with quinoa globulins, and to a lesser extent with globulins of sunflower and rice. Moreover, the anti-Gp serum was unable to detect either conformational or sequence epitopes in globulins of soybean, wheat, buckwheat, rice, and rye.     

Nutritional responses of rats to diets based on chickpea ( Cicer arietinum L.) seed meal or its protein fractions

The aim of this study was to isolate the protein fractions from chickpea, var. IAC-Marrocos, as well as to evaluate its in vivo nutritional protein quality. Among the proteins, albumins showed better nutritional value in the in vivo assays and amino acid contents, despite their higher trypsin inhibitor contents. Trypsin inhibitors were found to be heat labile in all samples, but the digestibility results for unheated and heated flour and albumins suggest that their contents are not very decisive. The PER values for casein (not supplemented) were very similar to those of heated flour and unheated or heated albumin and total globulins. The albumin and glutelin fractions showed the best results for PDCAAS, however, lower than those of casein. Despite the high digestibility of the globulin the very low essential amino acid content lowered its PDCAAS, and it had the lowest values.     

Environmental conditions during vegetative growth determine the major proteins that accumulate in mature grapes

Despite recent reports in the literature that chitinases comprise approximately 50% of the soluble proteins present in mature Vitis vinifera L. (cv. Moscatel) grapes, with the other major proteins being thaumatin-like proteins, a careful inspection of the published data reveals inconsistencies as to which proteins actually accumulate in mature grapes. Mature Moscatel grapes were harvested in the same vineyard in two consecutive years, 1999 and 2000. The grapes exhibited widely distinct polypeptide patterns when analyzed by either FPLC cation exchange chromatography or two-dimensional electrophoresis: whrereas the 2000 grapes possessed a much higher protein content (1.96 versus 1.11 mg g(-)(1) of fresh weight), the 1999 grapes contained a greater heterogeneity of polypeptides. In addition, highly specific antibodies that recognize the pathogenesis-related proteins present in the grapes demonstrated that the 2000 harvest grapes had a wider variety of pathogenesis-related polypeptides. N-Terminal sequencing of the major polypeptides revealed differences in the relative abundance and amino acid sequence of thaumatin-like and osmotin-like polypeptides present in the 1999 and 2000 harvest Moscatel grapes and did not detect the presence of chitinase. As a whole, the data indicate that the expression and accumulation of the major proteins in grapes is essentially determined by the environmental and/or pathological conditions prevailing during grape development and maturation. The potential physiological and technological implications are discussed. The results of the present work suggest that it is not possible to base varietal differentiation of grapes on the profile of the pathogenesis-related polypeptides present in the mature berries.     

Characterization of methanol-soluble and methanol-insoluble proteins from developing peanut seed

The 85% methanol-soluble proteins are known to specifically contribute to the production of flavor of roasted peanut. To determine the nature of the 85% methanol-soluble proteins, they were isolated from the peanut seed, and the 85% methanol-soluble (MS) and 85% methanol-insoluble (MIS) fractions were characterized using polyacrylamide gel electrophoresis (PAGE) and capillary electrophoresis. The results showed that the 85% MS fraction contained lower amounts (9-10%) of protein than the MIS fraction (15-33%). Protein content of the MIS fraction increased more significantly during seed maturation than it did in the MS fraction. Unlike the protein, free amino acids and soluble sugars levels of the MS fraction decreased significantly during seed maturation. The 85% MS fraction contained predominantly low molecular weight (<20 kDa) proteins/polypeptides, whereas the MIS fraction contained a mixture of polypeptides with molecular weight between 14 kDa and 90 kDa. SDS-PAGE showed no major changes in the polypeptide composition of the MS fraction during seed maturation. Capillary electrophoretic analysis revealed major qualitative and quantitative changes in the protein and polypeptide composition of the MS and MIS fractions during seed maturation. Fatty acid analysis of these fractions indicated that the MS fraction is lipoprotein in nature and rich in oleic and linoleic acids.     

Characterization of Protein Fractions of Rice Bran to Devise Effective Methods of Protein Solubilization

Proteins from the defatted brans of representative rice cultivars were fractionated into albumins, globulins, prolamins, and acid-soluble glutelins, accounting for 34, 15, 6, and 11% of the total bran proteins, respectively. The remaining insoluble residue protein, after treatment with 0.1M sodium hydroxide, resulted in the solubilization of 95% of the residue protein, representing 32% of the total bran protein. The relative molecular mass (M_r) values determined by size-exclusion HPLC were 10–100 kDa, 10–150 kDa, 33–150 kDa, and 25–100 kDa for the fully dissociated polypeptides of albumins, globulins, prolamins, and acid-soluble glutelins, respectively. Despite a breakdown of disulfide bonds of the residue protein during sodium hydroxide solubilization, the M_r of the majority of the fully dissociated polypeptides of this fraction ranged from 45 to 150 kDa. Insolubility of residue protein was due mainly to its strong aggregation and extensive disulfide bond cross-linking. Efficient methods may be developed for solubilizing up to 98% of rice bran protein by the use of dissociating and disulfide breaking agents currently in use in the food industry.     

Environmentally induced changes in amino acid composition in the grain of durum wheat grown under different water and temperature regimes in a Mediterranean environment

Amino acid composition is an important feature in determining the nutritional value of wheat grain for human and animal diets. Environmental conditions are known to influence protein quantity as well as grain production and, in turn, amino acid composition. In this study, grain yield, protein content, and amino acid composition were determined in 10 durum wheat genotypes under three water and temperature regimes in a Mediterranean environment. The highest value for grain-protein content (15.7%) was found in the warmer and driest environment and the lowest (12.8%) in the irrigated environment. Although amino acid composition showed significant variation for all genotypes, with the exception of arginine and cysteine, major changes in amino acid composition were caused by environmental conditions and in particular by water availability and temperature during the grain-filling period, which significantly altered the duration of grain development. The amino acids with the highest percentage of variation between environments were tyrosine (26.4%), lysine (23.7%), methionine (20.3%), threonine (19.3%), and valine (15.6%), whereas phenylalanine (5.1%), glycine (6.4%), and aspartic acid (6.8%) showed the least variation between environments. Whereas the content of glutamine, phenylalanine, and proline increased with the decrease in grain-filling duration, the remaining amino acids tended to diminish, presumably because high temperature and drought favored the deposition of gliadins (proteins particularly rich in glutamine and proline), to the detriment of albumins and globulins (proteins especially rich in threonine, lysine, methionine, valine, and histidine). Despite the negative correlations found between the percentage of protein and its content in essential amino acids, the results indicate that reductions in lysine per unit of food were not very pronounced (0.32 to 0.29 g/100 g of flour) with increases of up 22.7% in grain-protein content, whereas threonine did not change and valine even slightly increased.     

Characterization of the soluble allergenic proteins of cashew nut (Anacardium occidentale L.)

The allergens associated with cashew food allergy have not been well-characterized. We sought to identify the major allergens in cashew nut by performing IgE immunoblots to dissociated and reduced or nonreduced cashew protein extracts, followed by sequencing of the peptides of interest. Sera from 15 subjects with life-threatening reactions to cashews and 8 subjects who tolerate cashews but have life-threatening reactions to other tree nuts were compared. An aqueous cashew protein extract containing albumin/globulin was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and subjected to IgE immunoblotting using patient sera. Selected IgE reactive bands were subjected to N-terminal amino acid sequencing. Each of the 15 sera from cashew-allergic subjects showed IgE binding to the cashew protein extract. The dominant IgE-binding antigens in the reduced preparations included peptides in the 31-35 kD range, consistent with the large subunits of the major storage 13S globulin (legumin-like protein). Low-molecular-weight polypeptides of the 2S albumin family, with similarity to the major walnut allergen Jug r 1, also bound IgE. The sera from eight patients who tolerate cashew but displayed allergies to other tree nuts showed only minimal or no IgE binding to cashew. Cashew food allergy is associated with the presence of IgE directed against the major seed storage proteins in cashew, including the 13S globulin (legumin group) and 2S albumins, both of which represent major allergen classes in several plant seeds. Thus, the legumin-group proteins and 2S albumins are again identified as major food allergens, which will help further research into seed protein allergenicity.     

Characterization of globulins from common vetch (Vicia sativa L.)

The proteins from Vicia sativa L. (common vetch) seeds were investigated. Protein comprises approximately 11.4% of the seed fresh weight, >50.8% of which is composed by globulins and 43.6% by albumins. The globulins may be fractionated into two main components, which were named alpha-vicinin (comprising 73% of the total globulin fraction, and hence >37% of the total seed protein) and beta-vicinin. Two minor globulin components are also present, gamma-vicinin and delta-vicinin. alpha-Vicinin, the legumin-like globulin, with a sedimentation coefficient of 10.6 S, is a nonglycosylated, disulfide-bond-containing globulin, composed of a group of subunits with molecular masses ranging from 50 to 78 kDa. Upon reduction, each of these subunits releases a heavy polypeptide chain (34-66 kDa) and a light polypeptide chain (21-23 kDa). beta-Vicinin, the vicilin-like globulin, with a sedimentation coefficient of 7.7 S, is a nonglycosylated globulin that contains no disulfide bonds and consists of two major polypeptides with molecular masses of 58 and 66 kDa. gamma-Vicinin is a minor, glycosylated, disulfide-bond-containing globulin. In the reduced form, it comprises six polypeptide chains with molecular masses of 12, 19, 21, 22, 23, and 31 kDa. Finally, delta-vicinin is a minor, highly glycosylated globulin that exhibits hemagglutinating activity. It is composed of a major 47 kDa polypeptide and two minor (33 and 38 kDa) polypeptides. N-terminal sequencing of the delta-vicinin 47 kDa polypeptide revealed no homology to any other known storage protein.     

Characterization of the proteins from Vigna unguiculata seeds

The proteins from Vigna unguiculata (L.) Walp. (cowpea) seeds were investigated. Globulins constitute over 51% of the total seed protein, with albumins composing approximately 45%. The globulins may be fractionated by native electrophoresis or anion exchange chromatography into three main components, which were termed (in decreasing order of anodic mobility) alpha-vignin, beta-vignin, and gamma-vignin. alpha-Vignin, with a sedimentation coefficient of 16.5S, is a major, nonglycosylated globulin, composed of a major 80 kDa subunit, which upon reduction, produces two polypeptides (20 and 60 kDa). beta-Vignin, with a sedimentation coefficient of 13S, is a major, glycosylated globulin, composed of two main polypeptides (55 and 60 kDa) with no disulfide bonds. Finally, gamma-vignin, a minor globulin, is composed by one main type of subunit (22 kDa), which upon reduction, is converted into a single, apparently heavier polypeptide chain (30 kDa) due to the presence of an internal disulfide bond. Immunological analyses revealed structural homology between beta-vignin and beta-conglutin (the vicilin from Lupinus seeds) but not between alpha- or gamma-vignins and their Lupinus counterparts. Haemagglutination activity toward trypsinized rabbit erythrocytes was found exclusively in the albumin fraction and was strongly inhibited by N-acetylglucosamine or chitin.     

New insights into the structure of apolipoprotein B from low-density lipoproteins and identification of a novel YGP-like protein in hen egg yolk

Apoproteins of low-density lipoproteins (LDL) and soluble proteins (livetins) contained in hen egg yolk plasma have been demonstrated as being essential to the interfacial and emulsifying properties of yolk. The knowledge of their structure is necessary to better understand these properties. Purified protein fractions were separated by SDS-PAGE or 2D-PAGE and identified through the LC-MS/MS of their trypsin peptides. Hen blood apolipoprotein B gives rise to nine different apoproteins in LDL after maturation and proteolysis. Among these apoproteins, two protein fragments appeared to be less accessible to proteases and could be enriched in beta-sheets and firmly associated with lipids. Plasma soluble proteins were constituted by approximately 45% of yolk immunoglobulins with a high heterogeneity of the variable regions of both heavy and light chains, 41% of glycoproteins constituted by YGP42 and YGP40, 14% of albumins, and one new minor protein we called YGP30, showing 75% similarity to YGP40.     

Separation of Water‐Soluble Proteins from Cereals by High‐Performance Capillary Electrophoresis (HPCE)

Most research concerning grain proteins has concentrated on the gluten storage proteins. The albumins and globulins are the water‐ and salt‐soluble proteins that contain biologically active enzymes and enzyme inhibitors. A free‐zone capillary electrophoresis method was developed to separate these proteins. Optimization included sample extraction method, capillary temperature, buffer composition, and additives. The optimal conditions for separation of these proteins was 50 μm i.d. × 27 cm (20 cm to detector) capillary at 10 kV (with a 0.17 min ramp‐up time) and 25°C. The optimum buffer was 50 mM sodium phosphate, pH 2.5 + 20% acetonitrile (v/v) (ACN) + 0.05% (w/v) hydroxypropylmethyl‐cellulose (HPMC) + 50 mM hexane sulfonic acid (HSA). Sample stability was an issue that was addressed by lyophilizing fresh extracts and redissolving in aqueous 50% ethylene glycol and 10% separation buffer. This method was successfully used in both wheat flour and whole meal samples. Comparisons were made of several wheats of different classes as well as several cereal grains. This methodology could be useful in screening cereal grains for important enzymes and their impact on end‐use quality such as food functionality, food coloration, and malting quality.     

Improved Method for Isolating and Quantitating α‐Amino Nitrogen as Nonprotein,True Protein,Salt‐Soluble Proteins,Zeins, and True Glutelins in Maize Endosperm

The conventional Landry‐Moureaux method for selective extraction of maize proteins was modified by reducing the contact time of meal with extractants and by removing 55% 2‐propanol as extractant. The new procedure, coupled with a method for quantitating protein at microgram level, was used for assessing the nitrogen distribution of four soluble protein fractions present in 100‐mg samples of endosperm originating from six maize inbreds and opaque‐2 versions. Proteins extracted with 55% 2‐propanol plus reductant were made up of α‐, β‐, γ‐, and δ‐zeins. Proteins extracted subsequently with salt plus reductant were minor and poor in lysine (1 mol%).They were associated with zeins. Comparison of present data with those available in the literature showed a close similarity for a given genotype between the percentage of total α‐amino nitrogen extracted by 2‐propanol plus reductant than by salt plus reductant under conditions of the modified procedure and that of total Kjeldhal nitrogen extracted by 2‐propanol with and without reductant, and by salt plus reductant, using the conventional procedure. A simplified protocol was described and tested for isolating and quantitating α‐amino nitrogen as nonprotein, true protein, salt‐soluble proteins, zeins, and true glutelins in any sample of maize endosperm.     

Isolation and identification of two lipid transfer proteins in pomegranate (Punica granatum)

Lipid transfer proteins (LTPs) are a family of low molecular mass (7-9 kDa) polypeptides, the members of which share 35-95% sequence homology. These proteins are widely distributed throughout the plant kingdom and are receiving attention for their biochemical characteristics and biological activity. LTPs are indeed studied in different research fields varying from allergy to food technology, and numerous molecules belonging to this class are progressively being identified and investigated. Proteins from pomegranate juice were fractioned by cation exchange chromatography and analyzed by SDS-PAGE. Two proteins were identified as putative LTPs on the basis of their molecular weights and their electrophoretic behaviors under reducing and nonreducing conditions. Finally, proteins were purified and characterized by mass spectrometry. This analysis confirmed that the two polypeptides are LTPs on the basis of an amino acid sequence common to LTPs from other plant sources and cysteine content. The two proteins, named LTP1a and LTP1b, showed similar molecular masses but different immunological profiles when immunodetected with rabbit antibodies specific for Pru p 3 and human IgE from a patient suffering from pomegranate allergy. The demonstration of the existence of two immunologically unrelated LTPs in pomegranate confirms the variability and the complexity of the plant LTP family. This should be taken into account when the role of these proteins as elicitors of allergies to fruits is investigated and could help to explain the contradictory literature data on pomegranate allergy.     

干旱胁迫及复水对棉花叶片氮代谢的影响

以不同抗旱型棉花品种新陆早7号和新陆早24号幼苗为试材,研究持续干旱胁迫及复水对棉花幼苗叶片氮代谢关键酶活性、可溶性蛋白质、总氮和游离氨基酸含量的影响。结果显示:随着干旱胁迫天数的增加,新陆早7号和新陆早24号硝酸还原酶(NR)活性与对照相比分别降低 51.67%和74.22%;谷氨酰胺合成酶(GS)活性分别降低50.65%和72.95%;谷氨酸合成酶(GOGAT)活性分别降低 43.11%和68.36%;谷氨酸脱氢酶(GDH)活性在处理4d达到最大值;可溶性蛋白质含量分别增加了56.96%和34.12%;总氮含量分别增加了11.39%和8.43%;内肽酶活性分别增加了115.76%和165.06%;铵离子含量分别增加了92.29%和111.56%。干旱胁迫使新陆早7号在胁迫前期游离氨基酸含量增加慢,后期增加快;新陆早24号游离氨基酸含量的变化趋势与新陆早7号则相反。复水后,新陆早7号 NR、GS、GOGAT和GDH等活性恢复较快,内肽酶活性和游离氨基酸、铵离子、可溶性蛋白质和总氮含量下降也较快。试验表明,抗旱棉花品种具有较强的铵离子同化能力,增加渗透调节物质游离氨基酸含量,特别是脯氨酸含量增强其耐旱性。     

Proteome profiling of seed storage proteins reveals the nutritional potential of Salicornia brachiata Roxb., an extreme halophyte

Salicornia brachiata is an extreme halophyte that grows in salty marshes and is considered to be a potential alternative crop for seawater agriculture. Salicornia seeds are rich in protein, and its tender shoots are eaten as salad greens. Seed storage proteins were fractionated by sequential extraction using different solvents, including distilled water for albumins, NaCl (1.0 M) for globulins, NaOH (0.1 N) for glutelins, and ethanol (70% v/v) for prolamins. Globulins accounted for 54.75% of the total seed storage proteins followed by albumins (34.30%) and glutelins (8.70%). The fractionated proteins were characterized using 2D-diagonal SDS-PAGE and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. The globulin fraction, composed of seven intermolecular disulfide-linked polypeptide pairs of molecular mass 63.5, 62.5, 54.7, 53.0, 43.2, 38.5, and 35.1 kDa, encompassed a basic and an acidic subunit. Two-dimensional gels revealed approximately 32 spots, with isoelectric points and molecular masses ranging from 4.93 to 11.6 and from ～5.2 to ～109.4 kDa, respectively. Protein spots were identified by MALDI-TOF MS peptide mass fingerprint analysis and further classified. Homology analysis demonstrated that 19% of the proteins were involved in metabolism, 16% were involved in signaling, and 15% were regulatory proteins. Peptide mass fingerprint analysis confirmed the presence of inter- and intramolecular disulfide linkages in the globulin fraction. Sulfur-rich proteins are of high nutritional value, and disulfides make S. brachiata a potential source of dietary supplementation.