The degradation of lectins,phaseolin and trypsin inhibitors during germination of white kidney beans,Phaseolus vulgaris L.

White kidney beans (Phaseolus vulgaris), cv Processor, contain a relatively high content of phaseolin (storage protein), lectins and a special group of glycoproteins as well as a considerable amount of protein-type trypsin inhibitors. Protein digestion of raw Processor beans in monogastrics, for example pigs, is disturbed by poorly digested, phaseolin lectins, which can bind to carbohydrates in brush border membranes of the small intestinal epithelium, and trypsin inhibitors. The effect of the germination of white kidney beans on lectins, phaseolin and trypsin inhibitors was studied in order to achieve a degradation of lectins, phaseolin and trypsin inhibitors and an increase ofin vitro enzymatic hydrolysis of the protein of bean flour. Therefore, whole bean extracts were examined throughout a germination period of up to seven days for their lectin and phaseolin pattern, lectin content, binding capacities of functional lectins towards brush border membranes and trypsin inhibitor content. In addition thein vitro enzymatic hydrolysis by pepsin and pancreatin of the protein from flours of (un)germinated white kidney beans was studied. SDS-PAGE demonstrated a degradation of E-lectins and a disappearance of L-lectins and phaseolin during germination. Results indicated a decrease of the lectin content by 85%, a loss of binding capacities of functional lectins towards brush border membranes by 91%, and a decrease of trypsin inhibitors by 76%, in bean flour after germination for seven days. A maximum inin vitro enzymatic hydrolysis of protein from bean flour was already established after germination for half a day.     

Studies on α-amylase and trypsin inhibitors in legume seeds using agar diffusion and isoelectric focusing techniques

Amylolytic and tryptic inhibitors of faba bean extracts were determined by an agar diffusion test. The amylolytic inhibitor had protein characters. Furthermore, water-soluble trypsin inhibitors ofCicer arietinum, Lens esculenta, Lupinus termis, Phaseolus vulgaris, Pisum sativum, Trigonella foenum-graecum andVicia faba which were separated by polyacrylamide gel isoelectric focusing (PAGIF) in thin-layers, showed species specific patterns. Negative staining showed 10 bands for French beans, 9 for fenugreek seeds, 8 for lentils and chickpeas, 7 for peas and 6 for faba beans. Lupin seeds were free from trypsin inhibitors. Treatments (soaking, germination and heat processing) of faba beans reduced the number of trypsin inhibitors in PAGIF patterns, less after soaking and germination, but more after roasting and frying. No inhibitors were detected after cooking.     

Impact of microwave heating on hemagglutinins,trypsin inhibitors and protein quality of selected legume seeds

Selected legume seeds (dry and soaked) including faba beans, peas, chickpeas, soybeans, lentils and common beans containing 8 and 25% moisture, respectively, were subjected to microwave heating, and to a conventional cooking method to determine the heating effect on toxic compounds and protein quality. Trypsin inhibitors, hemagglutinins and available lysine were analyzed, and laboratory rats were used to determine digestibility and protein efficiency ratio (PER). Results indicated that microwaving destroyed trypsin inhibitors to a similar degree to that observed in beans cooked using the conventional method without affecting PER for raw seeds with low antinutrients content (faba beans, peas, chickpeas and lentils). Microwave-heated soaked soybeans had a higher amount of destroyed trypsin inhibitors, along with a higher PER, compared with microwave-heated dry soybeans. Microwave heating of common beans failed to destroy hemagglutinins and trypsin inhibitors, and consequently their digestibility and PER values were poor. Finally it was concluded that microwave heating constitutes an adequate method for destroying hemagglutinins and trypsin inhibitors without affecting protein quality of most legume seeds, except for common beans that despite of this process retained the antinutritional substances.     

Effect of germination time and temperature on the functionality and protein solubility of sorghum flour

Sorghum was germinated for different time (12, 24, 36 and 48 h) at different temperatures (25, 30 and 35 °C) and the changes in their nutritional and functional properties of germinated sorghum flour were assessed and compared with native sorghum flour. Germination inversely affects the crude protein, fat, fibre and ash content. A decrease in water absorption and swelling power and increase in oil absorption capacity was observed due to enzymatic starch modification as the germination duration progressed. Germination of sorghum increased the gel consistency while paste clarity was decreased as compared to native flour. Proteins were modified by action of enzymes during higher germination time and temperature conditions, which results in significantly higher protein solubility of germinated sorghum flour, which also result in enhancing the foaming and emulsifying properties of the flour. Lowest % synersis value and least gelation concentrations were observed in native sorghum has, which increased during germination and were highest in sorghum germinated for 48 h at 35 °C. Germination in overall can be used as low cost natural bio-processing technique for the preparation of modified flour with enhanced function properties without chemical modification or genetic engineering.     

Alternative Food/Feed Perspectives of an Underutilized Legume <Emphasis Type="Bold">Mucuna pruriens</Emphasis> var. <Emphasis Type="Bold">Utilis</Emphasis>—A Review

Mucuna pruriens var. utilis, an underutilized tropical legume has a nutritional quality comparable to soya beans and other conventional legumes as it contains similar proportions of protein, lipid, minerals, and other nutrients. The beans have been traditionally used as a food in a number of countries, viz., India, Philippines, Nigeria, Ghana, Brazil, and Malawi. Recently, the velvet beans are exploited as a protein source in the diets of fish, poultry, pig, and cattle after subjected to appropriate processing methods. Although the velvet beans contain high levels of protein and carbohydrate, their utilization is limited due to the presence of a number of antinutritional/antiphysiological compounds, phenolics, tannins, L-Dopa, lectins, protease inhibitors, etc., which may reduce the nutrient utilization. Unfortunately, even though many researchers all over the world working on Mucuna, only scanty and conflicting information are available regarding its utilization as a food/feed and no scientific gathering to date has focused on the food/feed applications of Mucuna. Hence, the present review has been emphasized on the nutritional potential of this underutilized, nonconventional legume and current state of its utilization as food/feed for both human beings and livestock throughout the world.     

Chemical composition and nutritional potential of yam bean seeds (Pachyrhizus erosus L. Urban)

Pachyrhizus erosus seeds were analysed for proximate composition, minerals, protein fractions, antinutritional factors, and rotenoids. The seeds showed a high content of proteins, lipids, Fe and Ca, in comparison to other legumes. Glutelins constitute the highest protein fraction, followed by globulins. Antinutritional substances detected as tannins, hemagglutinating activity and trypsin inhibitory activity, were in low concentrations. Seeds were also processed to obtain a flour which showed proper characteristics, good in vitro digestibility, significant rotenoid reduction level and amino acid composition rich in essential amino acids, except methionine.     

Involvement of carboxypeptidase in the degradation of the mung bean (Vigna radiata) trypsin inhibitor during germination and early seedling growth

Examination of the substrate specifity of the carboxypeptidase activity of ungerminated and germinated mung beans (Vigna radiata (L.) Wilczek) reveals the presence of two distinct enzymes. The first of these, carboxypeptidase I, is maximally active against carbobenzyloxy-Ala-Phe. It is present in large amounts in the cotyledons of ungerminated seeds, and declines rapidly during germination. The second, carboxypeptidase II, is most active against carbobenzyloxy-Phe-Ala. This enzyme increases in the cotyledons during germination and seedling growth.The possible involvement of one or both of these enzymes in the degradation of the native mung bean trypsin inhibitor (MBTI - F) and its proteolytically modified forms (MBTI - E, - C, and - E') during germination was also examined. The enzyme catalyzing one step in the conversion of MBTI - E to MBTI - C has been isolated with 557-fold purification from germinated mung beans. The inhibitor-degrading enzyme has a molecular weight of approximately 60,000. It is optimally active against MBTI - E at between pH 4.0 to 4.5. No activity was found with MBTI - F, - E', or - C as substrates. The enzyme has been identified as the carboxypeptidase II on the basis of (1) coelution during chromatography, (2) action on synthetic and natural substrates, (3) sensitivity to enzyme inhibitors, and (4) temporal variation during germination. The MBTI degrading carboxypeptidase removes the two carboxyl-terminal residues from MBTI - E to produce an inhibitor species that co-migrates with MBTI - C on polyacrylamide gel electrophoresis, but has not been subjected to the internal cleavage and proteolysis at the amino-terminus found in MBTI - C.Supported by National Science Foundation Grants PCM 8003854 and PCM 8301202Abbreviations used are: TNBS, 2,4,6-trinitrobenzenesulfonic acid; Cbz-, carbobenzyloxy-; PMSF, phenylmethylsulfonyl fluoride; MBTI, mung bean trypsin inhibitor; SDS, sodium dodecylsulfate; PAGE, polyacrylamide gel electrophoresis.     

Effects of germination and cooking on polyphenols and in vitro protein digestibility of horse gram and moth bean

Changes in polyphenols and in vitro protein digestibility due to germination and cooking were studied in horse gram and moth bean. Horse gram had higher amounts of polyphenols than moth bean. Polyphenol content in horse gram decreased from 1.6% to 1.1% at 48h germination, but in moth bean there was a reduction in polyphenol content from 1.3% to 1.0% at 24 h germination with a significant increase from 1.0% to 1.7% at 36 h germination. Seeds germinated for 48 h also exhibited higher amounts of polyphenols than in raw moth bean. The germination had no effect on total protein content, but a progressive increase in free amino acid content was observed in both legumes. In the ungerminated form, horse gram protein was more digestible than moth bean protein. The in vitro protein digestibility of both the legumes increased during germination. Cooking decreased the protein digestibility of horse gram either in ungerminated or germinated form, but such a decrease was noticed in moth bean at only 36 h germination. Moth bean either in raw form or germinated for 24 h did not exhibit a decrease in in vitro protein digestibility after cooking. Cooking decreased polyphenol content by 20%–50%. The apparent decrease in polyphenols in moth bean and horse gram due to cooking has been attributed to a decrease in assayable polyphenols.     

Effect of domestic processing,cooking and germination on the trypsin inhibitor activity and tannin content of faba bean (Vicia faba)

The effects of domestic processing methods like soaking for 12 h dehulling, ordinary cooking of whole as well as dehulled seeds at 151bs/inch² pressure for 15 and 25 min, and germination for 24, 36 and 48 h were observed on trypsin inhibitor (TIA) and tannin content of two varieties of faba bean viz. VH-131 and WF. A non-significant reduction in TIA was observed on soaking and dehulling, while tannins showed a reduction of 42 and 51% on soaking in VH 131 and WF, respectively. Dehulled seeds showed a decline in tannin content by 70 to 73% in two varieties. There was a significant reduction in TIA (75–76 per cent) and tannin (76–81%) on cooking, while autoclaving for 25 min almost completely eliminated both of these stress factors. Germination of seeds for 48 h led to a reduction of 64–65% in TIA and 90–91% in tannins, which was more than germination for 24 and 36 h. Soaked seeds gave positive results of both these antinutrients.     

Tannins,Trypsin Inhibitors and Lectin Cytotoxicity in Tepary (<Emphasis Type="Italic">Phaseolus acutifolius</Emphasis>) and Common (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis>) Beans

This study compared the levels of antinutritional components and cytotoxic effect of extracts, from tepary (Phaseolus acutifolius) and common (Phaseolus vulgaris) beans. Antinutritional factors were evaluated by determining their effect on the viability of epithelial cells isolated from rat small intestine. The protein and carbohydrates content were similar in all the genotypes studied (20 and 60%, respectively). Common beans presented higher content of trypsin inhibitors, tannins and lectins than tepary beans. There was not a significant correlation between tannins and cooking time. However, water absorption and cooking time correlated significantly (p < 0.05). Considerable variation was observed in lectin activity (1302–18161 Ul/mg) of extracts from different beans. Tannins, lectins, trypsin inhibitors and fat content differed between bean varieties whereas protein content was similar. The percent cellularity on rat epithelial cells was significantly different among protein extracts from different bean cultivars and ranged between 53.5% and 87.4% (p < 0.05). These results suggest that the incorporation of tepary beans in the diet would not alter the current nutritional contribution of common beans or introduce adverse toxic effects. The agronomic characteristics of tepary beans make them attractive for cultivation. However, the harder to cook phenomenon may be a limiting factor that needs further consideration.     

Effect of Germinated Soybean Protein Hydrolysates on Adipogenesis and Adipolysis in 3T3-L1 Cells

Germination of soybeans increases the bioavailability of some nutrients. An evaluation was done to determine if germination increased the anti-adipogenic and lipolytic effects of soybean. Soybeans were germinated for 0 to 6 days and protein concentrates extracted from beans germinated at each period. Soy protein concentrates can retain notable amounts of phytochemicals with anti-adipogenic activity. For this reason, it was evaluated the effect of protein hydrolysates with and without phytochemicals in the adipocyte-like cells after 3T3-L1 (murine fibroblasts) cell line differentiation. Cell viability decreased with exposure to the germinated soybean protein hydrolysates during the differentiation stage, but not during the fibroblast or mature adipocyte stages. Adipogenesis and triglycerides accumulation were strongly inhibited by the hydrolysate from soybeans germinated for 2 days (with ethanol-soluble phytochemicals), when compared to ungerminated soybean. Adipolysis increased with exposure to hydrolysates from beans germinated for 2 days (with phytochemicals) and 5 days (without phytochemicals). Germinated soy protein hydrolysates had an effect on inhibition of lipid storage in adypocites and increasing lipolysis, which was improved by changes of the protein and increased phytochemical content due to germination.     

Ukrainian Dietary Bread with Selenium-Enriched Soya Malt

The production of bread with addition of selenium-enriched soya malt was studied. Processing of this soya malt included soaking of the soya beans in the solution of hydroselenite with concentration 1.5 mg Se/L (20 μg of Se per 1 g of soya beans), then 4 days of beans germination at 20 °C, drying at 50 °C until moisture content 8%, separation from the sprouts and grinding. The soya malt was a powder containing 15–18 μg of Se in 1 g. The accumulated selenium was mainly in the protein fraction of soya malt. Addition of selenium-enriched soya malt to leaven intensified activity of yeasts and lactic acid bacteria. The quality of the wheat bread with selenium-enriched soya malt was better than that of the bread in control. The enriched bread had specific pleasant smell and soft texture. The daily intake of 277 g of bread with the selenium-enriched soya malt, which is added in quantity of 1.0–1.75% to mass of plain flour, ensures the consumption of 30–50% of selenium recommended daily allowance for 17 million population of the northern and northwestern Ukraine.

     

Preparation and antinutritional characteristics of dry bean (Phaseolus vulgaris L.) protein concentrates

Protein concentrates and starches were prepared by a wet extraction process from five dry bean (Phaseolus vulgaris L.) cultivars. The protein contents ranged from 69.7–76.4%. Concentrates prepared from dehulled beans under similar conditions had higher protein contents (80.6–87.9%). Each additional washing of the concentrates with distilled water increased their protein content. However, the protein recovery progressively decreased. The yield of starch ranged from 48.0–51.1% of the starting material. The solubility of bean proteins was minimal at pH 4.0, and under alkaline conditions, it was influenced by the tannin contents of the concentrates. Protein concentrates had lower trypsin, chymotrypsin, and amylase inhibitory activities as well as lower phytic acid and tannin contents compared to whole bean flours.     

Nutritional and antinutritional components of Canavalia spp. seeds from the west coast sand dunes of India

Seeds of two coastal sand dune wild legumes, Canavalia cathartica and Canavalia maritima from the west coast of India were analyzed for their nutritional and antinutritional properties. The seeds contained 35.5 and 34.1% crude protein, 52.8 and 50.5% crude carbohydrates, 1.3 and 1.7% crude lipids and 3.1 and 3.5% ash content, respectively. Among the minerals, potassium was the highest followed by phosphorus. Globulins and albumins constituted the major seed proteins of both plants. Most of the essential amino acids were at adequate levels. Although the crude lipids were below 2%, the unsaturated fatty acids were high. Seeds did not contain tannins and trypsin inhibitors. The levels of total phenolics were low, but showed high hemagglutination activity. The nutritional properties of these seeds have been compared with other Canavalia spp. and wild unconventional or tribal legumes. Overall, the study results suggest that the seeds of C. cathartica and C. maritima possess potential nutritive value particularly higher protein, essential amino acids and low levels of antinutritional factors. These wild legumes are adapted to low nutrient, high saline and drought conditions of the coastal sand dunes; their conservation and domestication are warranted as potential alternative sources of protein.     

Composition,functional properties,and biological evaluation of a plastein from cassava leaf protein

Rats lost weight continuously in a feeding trial to evaluate biologically a cassava leaf protein concentrate (LPC). The crude LPC had a high tannin content, 2.2%; 47% protein, and high levels of chymotrypsin and trypsin inhibitors, 27000 and 29500 units of inhibition, respectively. The application of the plastein reaction to the crude cassava LPC yielded a product with 90% protein, brought about a tenfold reduction of the tannin content, and lowered the trypsin and chymostrypsin inhibitors concentration to a non-detectable level. The cassava plastein showed the following value for the biological parameters: PER=2.2, NPR=3.4, TD=82.5%, BV=73.6, and NPU=60.7. The cassava LPC has a deep green colour, is bitter and astringent, and is insoluble in water. The plastein has a bland taste, a pale cream colour, is soluble in water, and has a high capacity of forming emulsions which are stable to thermal processing.     

Composition and functional properties of protein isolates obtained from commercial legumes grown in northern Spain

Pea (Pisum sativum), faba bean (Vicia faba) and soybean (Glycine max) seeds were characterized, and protein isolates were prepared following an isoelectric point precipitation procedure. Soybean seeds showed the highest protein content (36.7%) and carbohydrate was the major constituent in the pea (59.4%) and the faba bean seeds (52.1%). Protein contents were higher than 80% in all the protein isolates. The amino acid contents in the protein isolates were, in general, higher than those in their own starting seeds. The antinutritional factor contents were reduced after the protein isolate preparation. The highest reductions achieved for tannins were 95% in the faba bean protein isolate, and for phytates (45%) and trypsin inhibitor activity (46%) in the pea protein isolate. Haemagglutinating activity was not detected in any of the protein isolates. Minimum solubility values were observed at a pH range between 4.0 and 6.0, and maximal solubilities were obtained at basic pH values. The faba bean protein isolate showed the highest water and oil absorption capacities, and the best gelling properties. The soybean protein isolate had the best foam expansion capacity. Thus, the protein isolates had an improvement in some of the characteristics compared to their original seeds with lower contents in tannins, phytates and haemagglutinating activity, but had weak functional properties.     

Total Phenolic and Phytosterol Compounds and the Radical Scavenging Activity of Germinated Australian Sweet Lupin Flour

In addition to their favourable nutritional profile, legumes also contain a range of bioactive compounds such as phenolic compounds and phytosterols which may protect against chronic diseases including cancer and cardiovascular disease. Germination of some legume seeds has been previously reported to increase the concentration of the bioactive compounds. In this study, the effect of germination of Australian Sweet Lupin (ASL) seeds for 9 days on the concentration of some bioactive compounds and the radical scavenging activity in the resulting flour was determined. The concentration of total phenolic compounds in methanolic extracts of germinated ASL flour was determined using Folin Ciocalteu reagent and phytosterols in oil extracts were analyzed by gas–liquid chromatography. The methanolic and oil extracts were also used to determine radical scavenging activity toward 2,2-diphenyl-1-picrylhydrazyl. In the methanolic extracts of germinated ASL flour, phenolic contents and the antioxidant activity were significantly increased following germination (700 and 1400 %, respectively). Analysis of the oil extracts of germinated ASL flour revealed that the concentration of phytosterols and the antioxidant activity were also increased significantly compared to ungerminated ASL flour (300 and 800 %, respectively). The relative proportion of phytosterols in germinated ASL flour was: β-sitosterol (60 %), stigmasterol (30 %) and campesterol (10 %). Germination increases the concentration of bioactive compounds and the radical scavenging activity in the germinated ASL flour.     

The presence and inactivation of trypsin inhibitors,tannins, lectins and amylase inhibitors in legume seeds during germination. A review

During the germination of legume seeds, enzymes become active in order to degrade starch, storage-protein and proteinaceous antinutritional factors. The degradation of storage-protein is necessary to make peptides and amino acids available in order to stimulate seed growth and early plant growth. Proteinaceous antinutritional factors such as amylase inhibitors, lectins and trypsin inhibitors are present in legume seeds and protect them against predators. However, during germination, they degrade to a lower level by the action of several enzymes. The effect of germination on the content and activity of amylase inhibitors, lectins, tannins and trypsin inhibitors is discussed.     

Comparison of the Chemical Composition and Nutritional Value of <Emphasis Type="Italic">Amaranthus cruentus</Emphasis> Flour and Its Protein Concentrate

Plants constitute an alternative source of proteins in the human diet, with advantages over animal proteins because of their low content of saturated fats and absence of cholesterol. Within the framework of a wider research project on the role of Amaranthus cruentus (Ac) in lipid metabolism, in this work the chemical composition and biological value of the Ac flour and its protein concentrate were compared. Proximate chemical composition, amino acid and fatty acid profiles, some antinutrient factors, and biological values were determined for Ac seed flour and its protein concentrate obtained by extraction at pH 11 and precipitation at pH 4.5. The flour protein content was 16.6 g% while that of the concentrate was 52.56 g%. The content of the soluble dietary fiber with a hypolipemic function was notably higher in the protein concentrate (12.90 g%) than in the seed flour (4.29 g%). The protein concentrate also exhibited a higher content of insoluble dietary fiber. The Ac flour and the concentrate contain 75.44 and 56.95% unsaturated fatty acids, respectively. Squalene, which affects the biosynthesis of cholesterol, was detected both in the flour and the concentrate oils, with a higher content in the concentrate (9.53%) as compared to the flour (6.23%). Comparison of the amino acid composition with the FAO pattern protein indicated that the concentrate does not have limiting amino acids, while the flour has leucine, threonine, and valine. The content of lysine was high in both the flour and the concentrate, making these products particularly useful as a complement for cereal flour, which is deficient in this amino acid. The biological quality analysis demonstrated an improvement in the quality of the concentrate. The presence of saponins, phytic acid, and trypsin inhibitors in the concentrate, which favor the metabolism of lipids, suggests that consumption of the concentrate might reduce the risk of heart disease.     

Physicochemical characteristics of flours of faba bean as influenced by processing methods

Wet- and dry-processing with and without heating treatments were used to dehull faba beans for preparation of flours from the cotyledons. Flour qualities were assessed by levels of tannin and trypsin inhibitor and other measures of proximate composition. High roasting temperature and shorter process time that improved the recovered cotyledon yields were verified significant by path analysis methodologies. In turn, the higher cotyledon recoveries correlated with higher protein levels and inversely with the measured tannin levels. Ash reductions were correlated to the wet processing options in hull removal while reductions in the insoluble dietary fiber were notably influenced by level of heating temperatures applied. Trypsin inhibitor levels ranged from 42 to 56% of the original with reductions tied to applications of wet and/or heat processing in each case improved by increased durations and temperatures of treatments.