Sweet lupine (Lupinus luteus, var. Aurea/Weico and Lupinus albus, var. Multolupa) proteins. I. Extraction and filtration by Sephadex

Samples of Lupinus luteus, var. Aurea/Weico and Lupinus albus var. Multolupa flours were analyzed. The flour proteins were extracted and fractionated by gel filtration, and the per cent pattern for both globulins and albumins was then determined. The dehulled seeds, previously analyzed for composition, were ground, defatted and consecutively extracted with distilled water (pH 5.0) and phosphate buffer (0.05 M, pH 8.5). The extracted protein content was measured by the Lowry simplified method. Globulins (pH 8.5 fraction) from both species were filtered through Sephadex G-100; besides, Lupinus albus globulins were filtered through Sephadex G-150, and absorbance of the collected fractions was measured at 280 nm. The dehulled seed (DS) of L. luteus and L. albus revealed a good protein content (58.2 and 41.0%, respectively). The protein extracted from L. luteus was constituted by 17% albumins and 58.4% globulins. In contrast, L. albus presented a higher albumin content (55.6%) than globulins (31.5%). The elution pattern for the Sephadex G-200 gel filtration showed for both lupine species analyzed a preponderant peak I corresponding in L. luteus to 55.0% and in L. albus to 84.1% of the total globulins content. From these results, it may be concluded that the dehulled seed protein content is 42.0% higher for the L. luteus sample than for the L. albus. The applied methodology indicated a predominant content of globulins above albumins in L. luteus, while in the case of L. albus, the albumin content was the highest.     

Characterization of corn proteins of the cultivars Venezuela-1, Arichuna, Obregon and Venezuela-1 Opaque-2

A study is presented on the protein composition of the Venezuelan corn cultivars Venezuela-1, Arichuna, Obregón and Venezuela-1 Opaque-2. The proteins were isolated and analyzed for their molecular weight and lysine and tryptophan content. Protein fractionation showed significant differences between normal corn cultivars and Opaque-2. The latter showed a low level of zein and a high content of alcohol-insoluble reduced glutelins (AIG), albumins and globulins in relation to the normal kernel. The relative increase of these protein fractions, rich in lysine and tryptophan, results in a higher concentration of these amino acids in the Opaque kernel, as revealed by the analysis of these compounds carried out in different cultivars. Electrophoretic analysis showed only small differences in the number and intensity of bands from the protein fractions of the maize cultivars under study.     

Electrophoretic study of albumins and globulins of 4 genotypes of Canavalia ensiformis

Polyacrylamide gel electrophoresis (PAGE) and PAGE-SDS were used to study seed albumins and globulins of Canavalia ensiformis. In PAGE the concentration of acrylamide used in the upper gel was 4.0% with a pH of 6.7 and in the lower gel 7.5% and 10% of acrylamide were used with a pH of 8.9. In PAGE-SDS the concentration of acrylamide was 4.4% with a pH of 6.8 in the upper gel and 7.5% and 12.6% with a pH of 8.8 in the lower gel. The material used were the Venezuelan genotypes Tovar, Yaracuy, Original and U-02. The albumins and globulins were extracted with a 0.5 M NaCl solution and then separated by dialysis against water and lyophilized. These protein fractions represented 84.85% of the total amount of protein in seeds. The albumins were separated in PAGE with 7.5% acrylamide into five fractions and globulins into six, with similar electrophoretic patterns between genotypes. In a similar manner, the patterns obtained in PAGE with 10% acrylamide were the same for all genotypes, showing five bands for albumins and three bands for globulins. With PAGE-SDS containing 7.5% of acrylamide, albumins were separated into as many as eight components, and globulins into as many as seven bands with mobilities between 0.2981 and 0.9932, with different patterns for each genotype. Also the patterns PAGE-SDS at 12.6% of acrylamide were different for the genotypes, separating proteins into a greater number of bands. The albumins showed as many as twenty-one bands with mobilities between 0.2603 and 0.7398, and globulins as many as sixteen bands with mobilities between 0.2454 and 0.7390. The PAGE patterns of the genotypes analyzed did not show differences between them. However, with PAGE-SDS different electrophoretic patterns were obtained which varied in the number and intensity of the bands, making it possible to distinguish the genotypes studied. The molecular weight of the albumins varied between 76,000 and 12,000 daltons and of the globulins between 80,000 and 12,000 daltons.     

Sweet lupine (Lupinus luteus, var. Aurea/Weico and Lupinus albus, var. Multolupa) proteins. II. Separation by electrophoresis

The albumins and globulins extracted from Lupinus luteus var. Aurea/Weico and Lupinus albus var. Multolupa defatted flour samples were analyzed by polyacrilamide gel electrophoresis. The purpose was to determine the percentage distribution of pattern in the electrophoretograms and the relative mobility values of protein. The L. luteus albumins densitogram showed six fractions, two of them, No. 2 and No. 4, represented 58.1% of the total. On the other hand, the globulins densitogram revealed five fractions, numbers 1, 2 and 3 representing 90.7%. The densitogram of peak I from Sephadex G-100 globulins filtration indicated two fractions similar to those number 1 and 2 of the total globulins, showing low relative mobility values (from 0.26 to 0.39). The L. albus albumins resolved in four fractions, with No. 2 being the most prevalent (54.3%). In regard to the L. albus globulins, these showed five fractions identical in distribution with those of peak I, fraction 2 appearing as the most important (48.2%). It was found that Sephadex G-100 did not perform a good separation. As to the relative mobility values of globulins, fractions 2 and 3 were the most prominent (relative mobility of 0.35 and 0.48, respectively). It may be concluded that L. luteus albumins presented more components but with lower relative mobility values than those of L. albus. As for the globulins from both lupine flour samples, the gel separated fractions were classified in two groups of components, those of high and of low relative mobility.     

Characterization of proteins from grain of different bread and durum wheat genotypes

The classical Osborne wheat protein fractions (albumins, globulins, gliadins, and glutenins), as well as several proteins from each of the four subunits of gliadin using SDS-PAGE analyses, were determined in the grain of five bread (T. aestivum L.) and five durum wheat (T. durum Desf.) genotypes. In addition, content of tryptophan and wet gluten were analyzed. Gliadins and glutenins comprise from 58.17% to 65.27% and 56.25% to 64.48% of total proteins and as such account for both quantity and quality of the bread and durum wheat grain proteins, respectively. The ratio of gliadin/total glutenin varied from 0.49 to 1.01 and 0.57 to 1.06 among the bread and durum genotypes, respectively. According to SDS-PAGE analysis, bread wheat genotypes had a higher concentration of α + β + γ-subunits of gliadin (on average 61.54% of extractable proteins) than durum wheat (on average 55.32% of extractable proteins). However, low concentration of ω-subunit was found in both bread (0.50% to 2.53% of extractable proteins) and durum (3.65% to 6.99% of extractable proteins) wheat genotypes. On average, durum wheat contained significantly higher amounts of tryptophan and wet gluten (0.163% dry weight (d.w.) and 26.96% d.w., respectively) than bread wheat (0.147% d.w. and 24.18% d.w., respectively).     

Fractionation and characterization of proteins from <Emphasis Type="Italic">Gevuina avellana</Emphasis> and <Emphasis Type="Italic">Rosa rubiginosa</Emphasis> seeds

Gevuina avellana and Rosa rubiginosa proteins were evaluated for their potential food use. The proteins were sequentially separated into five fractions according to their solubilities in deionized water, 0.5 M NaCl, 70% (vol/vol) isopropyl alcohol, 50% (vol/vol) glacial acetic acid, and 0.1 M NaOH. The five fractionated protein groups were then characterized by SDS-PAGE and gel filtration chromatography to determine their M.W. profiles. Ninety-six percent of G. avellana total protein was solubilized in three extraction stages, and 88% of R. rubiginosa total protein was solubilized in one extraction stage. Albumins were the major protein fraction in G. avellana and glutelins-1 the most abundant in R. rubiginosa. The protein solubility profile determined over the pH range 1–12 showed minimal solubilities at pH 3–5 and pH 3–7 for G. avellana and R. rubiginosa, respectively. Electrophoretic studies revealed the existence of proteins composed of two major kinds of polypeptides linked together via disulfide bonds and with molecular masses ranging from 13 to 119 kDa. Gel filtration chromatography profiles of globulins and albumins were studied for both seeds. Isoelectric focusing showed an isoelectric point in the ranges of 4.5–6 and 3–6.5 for G. avellana and R. rubiginosa proteins, respectively.     

Fractionation of Soybean Globulins Using Ca<Superscript>2+</Superscript> and Mg<Superscript>2+</Superscript>: A Comparative Analysis

The substitution of CaCl₂ by MgCl₂ was undertaken in Deak’s two-step process of separating the soybean 11S and 7S globulins, aiming at higher purities and lower phytic acid (PA) contents of recovered protein fractions. The effects of pH and the addition of NaCl were also evaluated. Compared with CaCl₂, MgCl₂ reduced the PA content of the 11S-rich fraction by 63–71% but increased that of the 7S-rich fraction by 14–28%, depending on pH. Correspondingly, more Ca²⁺ was recovered in the 11S-rich fraction, while more Mg²⁺ co-precipitated with the 7S-rich fraction. NaCl increased the purity of the 11S-rich fraction and reduced its PA content, but the purity of the 7S-rich fraction was reduced by using 50–100 mM NaCl. Lowering pHs from 6.4 and 4.8 to 5.6 and 4.0 in the two precipitation steps increased the yield of both fractions. The optimized fractionating procedure was as follows: the 11S-rich fraction was precipitated at pH 5.8 by using 5 mM MgCl_2, 10 mM NaHSO₃ and 20 mM NaCl, followed by the precipitation of the 7S-rich fraction at pH 4.5. The new method provided both fractions with satisfactory protein yields (22% for 11S and 16% for 7S), purities (88% for 11S and 80% for 7S) and PA contents (0.356% for 11S and 0.882% for 7S).     

Extraction and characterization of soluble protein fractions from Phaseolus lunatus L seeds

Legume proteins as a potential source of valuable nutrients, are the object of several studies in order to obtain the best use. A basic knowledge becomes more important for those proteins from species not wholly utilized, before using them as food ingredients. The objective of this work was to determine several structural and nutritional characteristics of the protein fractions from Phaseolus lunatus, separated in different solvents. The relative amount of extraction for the albumins (ALB), globulins (GLB), prolamines (PRL), and glutelins (GLT) was 62.3, 34.8, 1.4 and 1.5%, respectively. The SDS-PAGE electrophoretic profile of both ALB and GLB, showed seven common bands in intervals from 10 to 95 kDa, and 14 to 99 kDa, respectively; the amino acids profile showed that PRL was the rich fraction in sulfurated amino acids (11.5 g/100 g protein); the content of lysine in the fraction of ALB was smaller than expected but the requirement of the FAO in the fractions of GLB and GLT was covered. In general, the fraction of GLB had the best balance of amino acids and digestibility (80%); however, it had a relationship of calculated protein efficiency ratio (C-PER) of 0.11, smaller than the ratio in ALB (0.97). The calorimetric analysis showed denatured temperatures around 90 degrees C for the ALB, GLB, and GLU fractions. The PRL fraction probably did not present a thermal transition because the proteins were denaturalized by the extraction conditions.     

Evaluation of the contents and ratios of five fractionated proteins to elucidate the protein composition in soybean seeds

Selecting suitable soybean cultivars is important for food processing and exploring their endowment with desirable physiological functions. We applied the fractionation method previously established to compare soy protein composition among cultivars to promote such a selection. More than 95% of the proteins in soybean cotyledons were extracted from 13 soybean cultivars using a high-concentration salt solution. The extracted proteins were fractionated into five fractions, namely oil body-associated protein (OBAP), polar lipid-associated protein (PLAP), globulins (11S and 7S), and whey by centrifugation after tuning the solubility behavior of the proteins with various solutions. Protein species in each fraction were analyzed by SDS-PAGE. Protein content in the total extract and five fractions was quantified to characterize the protein composition of soybean cultivars. The correlation between the protein content of each fraction and the total protein in cotyledon was investigated. A strong positive correlation was found only for the 11S fraction (r = 0.82), followed by a positive correlation in the 7S fraction (r = 0.65). Thus, we surmised that the increased protein content in soybean was due to increased globulin content. Furthermore, the calculation of the average ratio of protein content in each fraction indicated the globulin fraction (7S and 11S) to be 52%, the lipophilic protein fraction (OBAP and PLAP) to be 33%, and the whey fraction to be 13%. The preparation method employed in this study is a promising tool for efficiently comparing the protein composition of soybean cultivars to evaluate the potential use of cultivars for food production.     

Effect of extrusion on the activity of anti-nutritional factors and in vitro digestibility of protein and starch in flours of Canavalia ensiformis

The effect of the extrusion (155 degrees C, 20% moisture, screw speed 75 rpm, feed speed 205 g min-1) on antinutritional factors of Canavalia ensiformis was studied. In vitro protein and starch digestibilities were assessed. The extrusion not affect protein content (23%) in the flours, but significantly (P < 0.01) decrease moisture content. The protein digestibility values were improved from 57.5 to 89.5%, these values were lower than casein (98.19%). The digestibility of starch values were improved from 37.7 to 53%. The protease inhibitors activities (trypsin and chymotrypsin) and alpha-amylase inhibitor activity were reduced by 95%. The haemagglutinating activity was eliminated as result of the high temperature employed during the extrusion process. The canavanine content in the flours were not affect by the treatment of extrusion.     

Metabolism and exudation of canavanine during development of alfalfa (Medicago sativa L. cv. verko)

The structural analog of amino acidl-arginine,l-canavanine (2-amino-4-guanidinooxybutyric acid), is found in 26 cultivars of alfalfa. Its concentration ranges from 6 to 16 mg/g of dry seeds. Canavanine represented more than 70% of the total soluble nitrogen in seeds. Practically all of the canavanine was stored in the cotyledons. Comparison is made of the canavanine content in the cultivars Verko and Europa harvested in different years. During sprouting, 29% of the guanidinooxy compound was translocated into the hypocotyl and radicle in 24 hr. In this early stage of seedling development, the level of the nonprotein amino acid, canavanine, increased threefold whereas the protein amino acid, arginine, as well as asparagine increased 11- and 35-fold, respectively. Two-day-old seedlings are capable of synthesizing canavanine derived from canaline up to 25%. Contrary to this finding in seedlings grown in the time range of 24 days, the guanidino compounds canavanine and arginine were metabolized rapidly, whereas asparagine increased. Furthermore, the toxic canavanine got into the environment of swelled seeds or into the rhizosphere of young seedlings and increased in the milieu to concentrations at 3–57M. In a biotest, this inhibited the growth of a tomato cell suspension culture as well as the growth of cabbage radicle.     

Pilot-plant fractionation of soybean glycinin and β-conglycinin

A laboratory process for separating glycinin and β-conglycinin from soybean flakes was successfully scaled up to the pilot-plant scale (15 kg soy flakes). Average yields of the glycinin and β-conglycinin fractions were both 9.4% on a dry basis (db). The protein contents of glycinin and β-conglycinin fractions were 92.8 and 97.7% db, respectively. The glycinin and β-conglycinin purities were 90.4 and 72.7% of the protein content, respectively, which were very comparable to those of the laboratory-scale process. The total sulfhydryl plus half cystine content of the glycinin fraction was 37.8 mol/mol protein and 14.8 mol/mol protein for the β-conglycinin fraction. The native glycinin structure loss in the glycinin fraction was negligible. The native β-conglycinin loss in the β-conglycinin fraction was 10%, as estimated by rocket immunoelectrophoresis analysis. Hydrophobicity index value showed that hydrophobic properties of the pilot-plant protein fraction were ordered, from high to low: β-conglycinin fraction > glycinin fraction > intermediate mixture fraction.     

Proteins from Crambe abyssinica oilseed. I. Isolation procedure

Crambe abyssinica is a promising new oil crop because of the specific properties of its oil. However, little information is available concerning the properties of the proteins, which constitute a major component of the seed. Therefore, a method for the isolation of proteins from Crambe seeds was developed. Protein extractability for whole and dehulled Crambe meal was studied as a function of pH. Highest extractability was obtained with dehulled meal at pH 11. Double extraction at this pH increased the extractability to about 66%. Protein precipitation from the above-mentioned extract was studied as a function of pH with and without addition of a precipitating agent, i.e., carboxymethylcellulose (CMC). Addition of CMC resulted in a protein recovery of about 75% at pH 4.4. Without CMC, about half of the protein was recovered by isoelectric precipitation. The remaining soluble protein could be concentrated by ultrafiltration with a recovery of about 65%. The developed process, not including CMC addition, results in two protein fractions, i.e., an isoelectric precipitate (protein content 75%) and a retentate (protein content 87%), which together account for about 50% of the protein present in Crambe meal. Application of heat decreased protein extractability, but the protein contents of the resulting fractions were comparable to those from non-heat-treated meal.     

Chemical and functional characterization of major protein fractions extracted from nontoxic Jatropha curcas byproduct meals

Jatropha curcas seeds are a suitable source of oil for biofuel, among other use. A protein-rich meal is obtained after oilseed extraction. The goals of this study were to determine the physicochemical and functional properties of a nontoxic genotype of J. curcas defatted meal (JCDM) and the seed storage protein fractions to identify future applications. Both glutelin and globulin were the predominant protein fractions obtained from JCDM (42.03 and 20.17 g/100 g of protein, respectively). Leucine, phenylalanine + tyrosine, and histidine content of JCDM and protein fractions met the Food and Agriculture Organization/World Health Organization recommendation for children. The protein solubility (PS) profiles showed minimum values (5.3%–59.7%) at pH 5–6 and maximum at pH 2 (79.7%–81.6%) and above pH 10 (84.6%–89.8%). These findings suggest that JCDM proteins could be used in the formulation of juice or protein-based beverages. All the proteins showed the highest values for foam expansion (231%–285%) at pH 9. JCDM and the albumin fraction formed highly stable foams at pH 9, while the globulin and glutelin foams were stable at pH 3 and 2, respectively. Protein with stable foams, like those from jatropha are suitable for application in ice cream, mousse, among others. The emulsion activity index had similar behavior as foam expansion, but did not follow a specific trend. Thus, the proteins are suitable for use in salad dressing, sausages, comminuted meats, and mayonnaise. Taken together, JCDM protein and its soluble protein fractions have strong promise as alternative proteins for food structuring.     

Amino acid composition of some Amaranthus sp. grain proteins and of its fractions

This study was carried out to determine the protein content of several Amaranthus sp. grains. Findings revealed this has a high lysine (5.3 to 6.3 of the protein) and sulphur amino acids content (3.4-4.0%), while leucine could well be limiting when those seeds are used as a sole protein source in food. Using the correction for in vitro protein digestibility, the chemical score varied from 50 to 67. The calculated protein efficiency ratios and biological values ranged from 1.39 to 1.80 and 53 to 68, respectively. Considering that amaranth grain is a good supplement to cereal grain, the protein of A. hypochondriacus HH5 (yellow seeds) and A. anclancalius (black seeds) was fractionated into albumin, globulin, prolamin and glutelin. The average proportions between those soluble proteins were 65:17:11:7, respectively. Albumin had the highest lysine content (7.3-8.2%), and globulin the highest methionine (4.1-5.3%) and phenylalanine (6.0-6.1%) content. Prolamin had the highest threonine (4.6-5.4%) and leucine (6.8-6.9%) content, while glutelin had a very low methionine content (0.6-1.0%). Based on the above-mentioned findings, the authors conclude the variation in the amino acid composition of the protein fractions can be used for genetic protein improvement.     

Biochemical and chemical partial characterization of Bauhinia forficata Link seeds

Seeds of Bauhinia forficata species were submited to biochemical characterization concerning fatty acids analysis, protein fractionization, and hemaglutinanting activity. The seed elementary analysis showed a high protein and lipids contents with 21.24% and 19.45% respectively. The more abundant fatty acid was linoleic acid with 46.47% of the lipidic fraction. With the exception of prolamins, the different proteic fractions (albumin, globulins, acid and basic glutelins) showed hemaglutinanting activity against rabbit red cells no treated and treated with proteolitic enzymes. The fraction acid glutelin showed the higher specific hemaglutinanting activity (1072.25 UH/mg P) against rabbit blood pre-treated with trypsin. Glutamin (16.20%) and Valin (11.07%) were the more abundant amino acids in the seeds. Therefore, B. forficata represent a possible optional source of food because exhibit a high energetic values.     

Effect of flotation on the dioxin distribution in size-fractioned fly ash of hospital solid waste incineration

After the flotation of hospital solid waste incinerator (HSWI) fly ash, two types of solid products, froths and tailings, are produced. This paper reports the effect of flotation on the dioxins distribution patterns in different particle size fractions (?25, ?38, +25, ?75, +38, ?106, +75 and +106 µm) of fly ash. The results showed that the froths had a higher small-size particle distribution than that of the raw fly ash. The dioxin content with particle size in the flotation products depended on the partition behavior of powder-activated carbon (PAC). The dioxin content in the fine particles of the froths was higher than that in the coarse particles, and the highest content was in the finest particles (?25 µm). The dominant dibenzo-p-dioxins and dibenzofurans (PCDD/F) congeners in the froths and tailings were similar to those in the raw fly ash. A positive correlation between carbon removal and PCDD/F removal with different size fractions was observed. The carbon removal efficiencies of the fractions of ?25 and ?38, +25 µm were evidently higher than those of the other particle size fractions. Similarly, the PCDD/F removal efficiencies in fractions of ?25 and ?38, +25 µm could reach 156.9% and 115.0%, respectively.     

Ebony (Phitecellobium flexicaule Benth) and proteins fractionation,solubilization, characterization and production of an isolate

Different combinations of pHs (2 to 12) and temperatures (25, 30 and 35 degrees C) were tested to obtain a protein isolate from ebony (Pithecellobium flexicaule, Benth) seeds. Seed proteins contained 54.6% albumins, 32% globulins, 5.7% glutelins and 1.3% prolamins. The isoelectric points for albumins, globulins and glutelins were in the pH range of 2.3-2.7. The average molecular weight of albumins ranged from 92 to 100 kDa and for the four globulin subunits in the range of 28.4 to 57.3 kDa. For isolate production, proteins were sequentially extracted with distilled water and a 5% NaCl solution. The resulting supernatants were mixed. The best extraction was achieved at pH 11 and 25 degrees C. 45.6% of the total seed protein was precipitated at pH 2.6 yielding an isolate with 90% protein (N x 6.25). The isolate contained high quantities of lysine, leucine, threonine and phenylalanine but were low in sulfur containing amino acids methionine and cysteine. The extraction process reduced tannins, phytates and trypsin inhibitor in 53, 70 and 70%, respectively. In vivo protein digestibility of the protein isolate was 85.4% and the corrected digestibility essential amino acid score was of 44% due to the lack of sulfur containing amino acids. In order to upgrade the protein quality of ebony isolate it is recommend to supplement with methionine or sulfur containing rich foods.     

Ultrasonic-assisted enzymatic extraction and antioxidant activity of polysaccharides from Setaria viridis

Ultrasonic-assisted enzymatic extraction (UEAE) and response surface methodology (RSM) were used to isolate polysaccharides from Setaria viridis (SVP). Optimal extraction conditions in the enzymatic hydrolysis process were: extraction duration, 68 min; extraction temperature, 51°C; ratio of enzyme to raw material, 1.6%; and ratio of liquid to raw material, 20 mL/g. Then, following ultrasonic treatment (180 W, 60°C, 60 min), the experimental yield was 8.94 ± 0.38%. Crude SVP was purified by DEAE cellulose-52 chromatography and Sephadex G-100 chromatography, resulting in the isolation of three fractions (designated SVP-1, SVP-2 and SVP-3). These SVPs were mainly composed of glucose residue, and SVP-3 had a significantly higher uronic acid content than the other two fractions. Additionally, all fractions showed strong antioxidant activities in vitro.     

Fractionation of sunflower seed proteins

Fractionation of sunflower seed salt-soluble proteins, which amount to nearly 80% of the total seed nitrogen, has been performed by a method we proposed in 1970 and which was confirmed by several others. Three varieties of seeds have been investigated: ‘Armavirec,’ ‘Peredovik’, and a pure strain. The occurrence of three groups of proteic fractions was confirmed. Their proportions, which fluctuate with varieties, are roughly: 20% for “light” (low molecular weight) albumins, 5–10% for “heavy albumins,” and 70–80% for globulins. The first group was isolated by Sephadex G-50 chromatography from the other two, which were separated by dialysis. A second chromatography of these three groups on Sephadex G-200 has been realized (with preliminarily calibrated columns for molecular weight evaluations). “Light” albumins appear as a rather homogeneous constituent with a molecular weight of 14,000 and an aminoacid composition showing high amounts of methionine, cystine, arginine and glutamine. “Heavy” albumins, which are still mixed with globulin fractions after dialysis, have a molecular weight of 48,000 and a very different aminoacid composition with a high level of lysine. Globulins are composed of at least four different fractions, two of which (M=12,000 and M=25,000) are presumably subunits of the other two and have significantly different aminoacid compositions.