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.     

Quick-cooking beans (Phaseolus vulgaris L.): I. Investigations on quality

Soaking three beans cultivars (Phaseolus vulgaris L.: Great Northern, kidney, and pinto) in mixed salt solution (sodium chloride 2.5% + sodium bicarbonate 1.5% + sodium tripolyphosphate 1.0% + sodium carbonate 0.5%) resulted in 80%–85% reduction in cooking time over corresponding controls. Irradiation (-rays) at 500 krads of soaked and dehydrated beans caused a reduction of nearly 50% in cooking time. Water uptake and leaching losses for each treatment during soaking at 22°, 37°, and 45°C were investigated. High temperature (37° and 45°C) and pH (9.0) caused greater water imbibition and total solid loss than at room temperature (22°C). Organoleptic evaluation revealed that quick-cooking Great Northern beans appear to be more acceptable than kidney and pinto beans. Quick-cooking cooked beans had better in vitro protein digestibility than conventionally cooked beans. Phenolic content was found to be inversely related to in vitro digestibility.Utah Agricultural Experiment Station Journal article no. 2481.     

Antinutritional factors in anasazi and other pinto beans (Phaseolus vulgaris L.)

Antinutritional factors of anasazi bean were compared to traditional pinto bean (Phaseolus vulgaris L.). Anasazi beans contained less (p<0.001) soluble and bound condensed tannins compared to pinto beans. No differences (p>0.05) in stachyose and raffinose content were found between the two bean types; verbascose was not detected at all. Significant (p<0.05) differences in lectin content were observed between anasazi and pinto bean. The lectins of anasazi beans were classified as non toxic and those of the pinto beans as toxic types. No differences (p>0.05) in inhibitor activity against human and bovine trypsin and chymotrypsin were found between the two bean types.     

The effect of slow-cooking on the trypsin inhibitor and hemagglutinating activities and in vitro digestibility of brown beans (Phaseolus vulgaris,var.Stella) and kidney beans (Phaseolus vulgaris,var.Montcalm)

Brown beans and kidney beans were subjected to two modes of cooking in a household slow cooker: (A), a fixed low setting for 10 h, and (B), a high setting for 2.5 h and a low setting for 7.5 h. Temperature changes in the beans were recorded. With treatment A over 90% of the hemagglutinating and trypsin inhibitor activities occurred after 6 h at which time the temperature had reached 80°C. With treatment B inactivation of these activities was almost complete at the end of 2 h when a maximum temperature of 100°C had been attained. The in vitro digestibility of the bean protein was considerably increased by either treatment. By way of contrast, only 20 min of heating was required to destroy these activities when the beans were brought to a boil in an open vessel.     

Effect ofCallosobruchus chinensis (Bruchid) infestation on antinutritional factors in stored legumes

Effect of bruchid infestation on selected antinutritional factors, saponin, phytic acid and trypsin inhibitor activities, of three (mung bean, chickpea and pigeon pea) legumes stored for six months was studied. The three antinutritional factors, saponin, phytic acid and trypsin inhibitor activity (TIA), increased with increased storage period. Saponin, phytic acid and trypsin inhibitor activity values were significantly lower in control samples compared to stored infested samples. The difference between the contents of antinutritional factors were significant (p<0.05) during different storage periods.     

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.     

Effects of heat treatments on trypsin inhibitor and hemagglutinating activities in winged bean

The effect of different heat treatments on inactivation of trypsin inhibitor and hemagglutinin of winged bean was investigated. Trypsin inhibitor extracted from winged bean meal was stable at 60 °C for 60 min. At 80 °C, the activity of the extracted inhibitor decreased by 25% within 5 min, and continued to decline gradually to a loss of 45% of activity after 30 min. When the extracted inhibitor was incubated at 100 °C, it exhibited a triphasic pattern of inactivation. The winged bean extract incubated at 60 °C lost 60% of its hemaggluinating activity within 30 min. At 80 °C, there was a complete loss of activity within 5 min. The microwave treatment to winged bean meal had no effect on trypsin inhibitor or hemagglutinating activities in the meal. However, infrared treatment to winged bean seeds for 60 seconds was effective in destroying most of the trypsin inhibitor and hemagglutinating activities. Autoclave treatment (120 °C at 15 lb pressure) for 10 min inactivated trypsin inhibitor and hemagglutinin in winged bean meal almost completely. Cooking of presoaked beans in boiling water for 30 min was effective in destroying most of the trypsin inhibitor and hemagglutinating activities.     

Comparative studies on the storage proteins and anti-nutritional factors from seeds ofPsophocarpus tetragonolobus (L.) DC from five South-East Asian countries

The seeds of 27 varieties ofP. tetragonolobus from six regions of South-East Asia have been examined by polyacrylamide gel electrophoresis. No marked variations in the electrophoretic patterns were found which could be exploited by plant breeders to improve nutritional quality with respect to sulfur containing amino acids. The amino acid compositions of varieties from the different regions showed little variation. Seed extracts of all 27 varieties ofP. tetragonolobus showed trypsin and chymotrypsin inhibitory activities, and hemagglutinating activities. The levels of trypsin inhibitory activities showed some genotypic differences varying from 22.2 to 42.5 mg trypsin inbibited g^?1 of seed meal. As a group the varieties from Malaysia showed the lowest levels of trypsin inhibitor. The chymotrypsin inhibitory activity also showed a similar variation (30.1–47.6 mg chymotrypsin inhibited g^?1 of seed meal). Seed extracts agglutinated type O⁺ and B⁺ human and rabbit erythrocytes with little difference in activity between varieties. Autoclaving winged bean seed meal prior to protein extraction inactivated the anti-nutritional factors and resulted in considerable reduction of protein solubility.     

Comparative study of enzymes related to proline metabolism in tepary bean (Phaseolus acutifolius) and common bean (Phaseolus vulgaris) under drought and irrigated conditions,and various urea concentrations

There are several mechanisms used by plants for survival in adverse environments such as drought, high temperature and salinity. The objective of this study was to evaluate the drought tolerance of tepary bean as a function of biochemical processes linked to isozyme synthesis and changes in enzymatic activity related to proline metabolism. Mature seeds of common beans var. flor de mayo, Phaseolus vulgaris and tepary beans Phaseolus acutifolius were grown under two water conditions (irrigation and drought), and four levels of urea. Vertical electrophoresis and spectrophotometric techniques were used to evaluate protein patterns, glutamate dehydrogenase (GDH), proline oxidase (PO) and pyrroline-5-carboxylate reductase (P5C reductase) enzyme activities. These enzymes were studied because they are directly related to protein synthesis. Electrophoretic patterns showed more proteins in tepary beans than in common beans with limited irrigation. GDH showed only one isozyme, with a molecular weight between 240 to 270 kDa. A decrease in PO activity was observed in common beans under drought stress with a value of 237 mol/min, in comparison to irrigation conditions of 580 mol/min. GDH and P5C reductase enzymes have had higher activity in common beans than in tepary beans under water stress. There was a significant difference only in glutamate dehydrogenase enzyme with respect to urea level. The results suggest that drought tolerance of tepary beans is due to biochemical processes related to proline metabolic enzymes.     

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.     

Enzyme inhibitors in tuber crops and their thermal stability

Tubers of Cassava (Manihot esculenta), yams (Dioscorea esculenta),aroids (Amorphophallus campanulatus, Colocasia esculenta, Xanthosoma sagittifolium) and Coleus (Solenostemon rotundifolius) were screened for inhibitory activities against amylase, trypsin and chymotrypsin. Coleus tuber possessed the highest anti-amylase activity, whereas Colocasia tuber was the most potent source of anti-tryptic and anti-chymotryptic activity. Xanthosoma tubers exhibited amylase inhibitory activity and Amorphophallus tubers antiprotease activity. Dioscorea esculenta had low levels of amylase and chymotrypsin inhibitors, while Cassava tubers were totally free of inhibitors. When tubers were processed by pressure cooking, there was significant reduction/complete elimination in inhibitory activity. Partial retention of inhibition was observed in the case of amylase inhibitor in Dioscorea, chymotrypsin inhibitor in Colocasia and trypsin inhibitor in Colocasia, Coleus and Amorphophallus. In vitro experiments on heat stability of the different inhibitors revealed almost similar pattern of inactivation.     

豆类中胰蛋白酶抑制剂活性测定与初步分析

大豆中主要的抗营养因子是胰蛋白酶抑制剂,在其它食用豆中也有存在。供试大豆中胰蛋白酶抑制剂活性(TIA)最高(20.30～26.30TIU/g),菜豆次之(2.93～6.41TIU/g),蚕豆、豌豆为2.01～2.87及1.66～2.38TIU/g。大豆中抑制剂活性与粗蛋白及粗脂肪之间存在显著负相关。测定结果表明:国内选育的不含SBTI—A_2基因大豆材料,抑制剂总活性为17.40TIU/g,低于其栽培品种亲本21.20～21.60TIU/g。     

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.     

Specific role of endopeptidases in modulating the nature of protein digestion products

The impact of various endopeptidases on the nature of protein digestion products was measured with the digestion cell technique. After a 30 min pepsin pre-digestion, casein and rapeseed concentrates were hydrolyzed with various amounts of pancreatin, trypsin and/or chymotrypsin. This hydrolysis was performed in a dialysis tube (molecular weight cut-off 1000) with continuous collection of the digested material. The addition of pure trypsin or chymotrypsin to pancreatin (Enzyme:Substrate 125) did not change the digestibility of casein. Only a higher pancreatin level (Enzyme: Substrate 112.5) increased the total protein digestibility without affecting the amino acid spectra. Rapeseed digestibility was markedly increased by the addition of pure trypsin to pancreatin. Lysine and arginine, target amino acids of trypsin, were favored at the expense of chymotrypsin and elastase target amino acids. Supplementation of pancreatin with, chymotrypsin enhanced rapeseed digestibility without affecting the relative amino acid digestibility. The impact of a higher pancreatin ratio (Enzyme: Substrate 112.5) was similar to that of enriched pancreatin but the rate of amino acid release was modified. The differences between protein sources were mainly attributed to protein structure.     

The effect of feeding soya bean trypsin inhibitor and repeated injections of cholecystokinin on rat pancreas

The effects of dietary soya bean trypsin inhibitor (SBTI) and of the daily injection of pure CCK or porcine intedstinal extract have been studied on the rat pancreas. The following parameters were measured: weight, DNA, RNA, trypsin, chymotrypsin, amylase, lipase, insulin and glucagon. The effects of dietary SBTI on the level of gastrin in the antrum have also been studied. Oral SBTI and i.p. CCK caused similar hypertrophy and hyperplasia but not the same activities of the enzymes. It is suggested that SBTI stimulates the secretion of at least two gastro-intestinal hormones: CCK and gastrin.     

Chemical changes during the fermentation of African locust-bean (Parkia filicoidea Welw) seeds for production of ‘Daddawa’

African locust-bean (Parkia filicoidea Welw) seeds were allowed to ferment under natural conditions and the chemical changes occurring during the fermentation were studied. Chemical analysis of fermented and unfermented beans showed a marked decrease in the total sugar content; crude and true protein levels all increased. The results of the chemical analyses indicate that fermentation resulted in protein enrichment of the fermented locust bean (daddaw). Fermentation also imparts a characteristic flavour and odour to the beans.     

Comparative study of the chemical composition of wild and cultivated beans (Phaseolus vulgaris)

Five wildPhaseolus vulgaris beans were compared with five cultivatedPhaseolus vulgaris beans in proximate composition, total (true) protein, amino acid composition, and toxic and antinutritional factors. The wild beans contained more protein (25.5% vs. 21.7%), ash (5.15 vs. 4.15%), crude fiber (7.08% vs. 5.04%) compared to cultivated beans while the former contained less fat (0.56 vs. 0.89%) and carbohydrates (61.64 vs. 68.05%). Sulfur amino acids were found to be limiting in both groups of bean as expected; however, the cultivated beans had a higher content of the limiting amino acids. Therefore, the cultivated beans showed a better amino acid profile than the wild beans. Toxic factors were not found in either type of bean; the determinations included saponins, alkaloids, and cyanogenic glycosides. The antinutritional factors investigated were hemagglutinins (lectins) and trypsin inhibitors. The wild beans presented a higher content of trypsin inhibitors (28 TUI per mg) and lectins (9.6) than the cultivated beans did (21 TUI per mg and 7 respectively). From the chemical point of view, domestication seems to be positive; however, the better protein nutritive quality of the cultivated beans should be further confirmed by biological assays.     

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.     

Toxische und Wachstumshemmende Faktoren in Bohnen

Zusammenfassung Gartenbohnen (Phaseolus vulgaris) und Helmbohnen (Dolichos lablab) enthalten toxische Phytohaemagglutinine, die bei Verfütterung der rohen, gemahlenen Bohnen zu Gewichtsverlust und Tod der Versuchstiere führen. Die in ihnen enthaltenen Trypsininhibitoren sowie der Amylaseinhibitor der Gartenbohnen haben keine nachweisbare Giftwirkung. Die von Trypsininhibitoren befreiten rohen Eiweiße aus beiden Bohnenarten sind in vitro gegen den Angriff von allen diesbezüglich geprüften Proteinasen sehr resistent und zeigen auch in vivo sehr geringe Verdaulichkeit. Das Zusammenwirken der erwähnten Faktoren wird für die Wachstumshemmwirkung der Hülsenfrüchte verantwortlich gemacht, doch kann das Vorhandensein von andern, bisher unbekannten, toxischen Inhaltsstoffen nicht ausgeschlossen werden.Bei Limabohnen (Phaseolus lunatus), die zwar das Wachstum hemmen, aber keine Letalwirkung zeigen, konnten ähnliche Befunde über die Verdaulichkeit und die nicht toxische Wirkung des Trypsininhibitors aufgezeigt werden, jedoch fehlen Hinweise auf eine mögliche Toxizität des in ihnen enthaltenen Agglutinins.

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Summary Both, kidney beans (Phaseolus vulgaris) and field beans (Dolichos lablab) contain toxic phytohemagglutinins. Ingestion of the raw, ground beans cause weight loss and death of the experimental animals. The two bean species also contain trypsin inhibitors and the kidney beans also an amylase inhibitor, which are devoid of detectable toxicity. Proteins from both bean species, even when free of trypsin inhibitors, are resistant against in vitro attack of all proteases tested and also show a very low in vivo digestibility. It is concluded that this property, together with the toxic haemagglutinins are responsible for the growth inhibition, but the existance of unrecognized toxic constituents can not be ruled out.Limabeans (Phaseolus lunatus) show growth inhibiting action, but their ingestion is not lethal. Their trypsin inhibitor is not toxic, but digestibility is also low. Nothing is known about a possible toxic action of their agglutinin.

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Resumen Las semillas de frijoles (Phaseolus vulgaris) y de gallinazo (Dolichos lablab) contienen fitohemagglutininas tóxicas. El consumo de estas semillas crudas y molidas produce pérdida de peso y la muerte de animales experimentales. Ambas especies de leguminosas contienen además inhibidores de la tripsina y en los frijoles se encuentra tambien un inhibidor de la amilasa pancreática. Estos últimos factores no exhiben, sin embargo, actividad tóxica significante. Las proteinas de ambos tipos de semillas son poco atacadas in vitro por enzimas proteolíticos, ni tampoco si se separan previamente de los inhibidores trípticos. Su digestibilidad in vivo es igualmente baja. Se concluye que esta característica, junto con la presencia de las hemagglutininas tóxicas son responsables, por lo menos parcialmente, de la toxicidad de las semillas crudas, aunque no se excluye la posibilidad de que esten presentes otros factores toxicos todavia no reconocidos.Habas de Lima (Phaseolus lunatus) tambien retardan el crecimiento de animales experimentales, si se incorporan en la dieta, pero su ingestión no causa la muerte. Contienen un inhibidor tríptico no tóxico y una hemagglutinina cuya acción biológica se desconoce.

     

Effects of soaking,cooking and fermentation on composition,in-vitro starch digestibility and nutritive value of common beans

A common bean variety, grown in Burundi, was either fermented, soaked and/or cooked, and then assessed for nutrient composition, in-vitro starch digestibility and protein nutritive value. A decrease in ash, most minerals, vitamins, and some essential amino acids was noted for soaked, cooked and soaked-cooked beans. Compared to untreated beans, soaking decreased soluble sugar (9.8 percent) but increased starch (7.3 percent) and soluble fiber (16.9 percent). In cooked beans, an increase in soluble sugar (1.5 percent), and a decrease in thiamine (81.7 percent), starch (24.6 percent) and soluble fiber (16.6 percent) and nitrogen (2.9 percent) contents were observed. Crude fiber (6.9 percent) and starch (10.0 percent) increased while fat (17.6 percent), fatty acids (linoleic: 10.7 percent; linolenic: 14.3 percent) and soluble sugars (25.4 percent) and nitrogen (14.4 percent) decreased in soaked-cooked beans. Fermentation increased potassium (11.6 percent), soluble fiber (18.9 percent), and some amino acids but decreased fatty acids (linoleic: 13.5 percent; linolenic: 19.9 percent), soluble sugar (75.2 percent) and vitamin (riboflavin: 41.0 percent; niacin: 24.5 percent) contents in common beans. However, the in-vitro starch digestibility was greatly improved (12.3 percent) by cooking while it decreased in soaked beans (29.2 percent). Soaking-cooking and fermentation did not have any significant effect on the digestibility of common bean starch. Finally, among the five treatments applied to common beans, only fermentation showed a significant improvement (8.3 percent) on the protein nutritive value of this legume.