Anti Nutritional Factors

Plant Foods for Human Nutrition 37:201-228 (1987) © Martinus Nijhoff Publishers, Dordrecht - Printed in the Netherlands 201

Anti-nutritional and toxic factors in food legumes: a review

Y.P. GUPTA Division of Agricultural Biochemistry, Indian Agricultural Research Institute, New Delhi-l lO012, India

Received 4 November 1985; in revised form 17 December 1986

Key words: proteolytic inhibitors, phytohemagglutinins, lathyrogens, cyanogenetic compounds, favism

Abstract. A comprehensive review on the presence of certain important anti-nutritional and toxic factors in food legumes has been conducted. These substances include proteolytic inhibitors, phytohemagglutinins, lathyrogens, cyanogenetic compounds, compounds causing favism, factors affecting digestibility and saponins. These factors are shown to be widely present in leguminous foods which are important constituents of the diet of a large section of the world's population, and particularly, of people in the developing countries.

Introduction

Food legumes constitute an important part of the diet of a large section of the population in the developing world, as a good source of proteins, carbohydrates, minerals and B-vitamins. But in the raw state, they contain certain toxic substances which include trypsin inhibitors, phytohemagglutinins, lathyrogens, compounds causing favism, cyanogenetic and goiterogenic factors, saponins and alkaloids [142, 146, 228]. It is reported [86, 142] that these substances are generally eliminated by soaking and subsequent discarding of the liquid and/or by heat treatment at relatively elevated temperatures. Some of these important substances have been comprehensively reviewed in this paper.

Factors affecting digestibility

Pulse digestibility has been found to be affected by the methods of processing and cooking, by the quantities consumed and by the state of the digestive tract. The presence of saponins, glycosides, tannins, alkaloids, conjugates of protein with phytin or hemi-cellulose and substances inhibiting the action of digestive enzyme trypsin in different food legumes adversely affect their digestibility as these substances are indigestible or are antagonistic to digestion [87].

Digestibility of food legumes has been consistently 15-20 per cent lower than that of casein [214]. The lower apparent protein digestibility is attributed to the quantities of tannins and other polyphenols present in the seed-coat [60]. The unfavourable influence of tannins on the nutritional properties of dry beans has

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been described earlier [193, 194]. Protein utilisation is also influenced by hemagglutinins and other inhibitors. Plant fibre interfered with protein digestibility [128]. The physiological factors affecting protein digestibility have been described by Kies [127].

Imbalance of amino acids and their availability also affected protein utilisation [87]. In vitro studies have shown that raw kidneybean is less digestible by pepsin and trypsin than casein, and autoclaving did not improve its digestibility by pepsin. But Chatterbuck et al. [33] observed that autoclaving markedly improved in vitro digestibility of soy protein concentrates and isolates. It was also found that different varieties of soybean showed wide variation in their in vitro digestion with trypsin [81]. In vitro pancreatic digestion of proteins in bengalgram, greengram, blackgram, redgram, lentil and pea was much slower than that of meat meal and casein [88]. Earlier, Manage & Sohonie [157] reported similar findings with horsegram that the rate of in vitro proteolytic digestion was slower as compared to that of casein. It seems that trypsin inhibitor has been interfering with proteolytic digestion. Similar. results were obtained with doublebean and fieldbean [105, 188].

Protein digestibility of pea, fababean and lentil meals and their concentrates and isolates (81 to 90 per cent) was found similar to that of casein (87 per cent), and their poor growth-promoting ability was reported to be due to growth- depressing factors such as tannins, trypsin inhibitors and hemagglutinins [17]. It was recently demonstrated [253] that the rate of protein digestibility in the raw bean extract was less than that of casein and bovine serum albumin due to the presence of hemagglutinins. It was suggested that lectins affected the activity of digestive enzymes. Protein digestibility of cooked pigeonpea meal was also found to be low [231]. In vitro proteolytic digestion of denatured protein phaseolin from Phaseolus vulgaris seeds showed that it was fully digested and its native form was partially digested [25]. The poor digestion in case of native form was due to the presence of trypsin inhibitor. It was reported that trypsin inhibitor and in vitro protein digestibility are related to nutritional quality [203, 223]. Deshpande et al. [47] also obtained poor in vitro proteolytic digestibility of Phaseolus vuIgaris dry seeds. Studies by Hernandez-Infante et al. [99] showed that in vitro digestibility of black beans when dried with broth was poor as compared to that obtained with white beans. In vitro studies on the digestibility of carbohydrates showed that greengram was digested at a faster rate than bengalgram, blackgram and redgram [87].

Germination and cooking were reported to improve in vitro protein digestibility of horsegram and mothbean [224]. E1Faki et al. [57] also observed improvement in in vitro protein digestibility of horsegram and cowpea by germination and cooking but there was no significant improvement in case of chickpea. Phillips et al. [189] found that cooking initially increased in vitro protein digestibility of cowpea and then decreased. Gross [79] reported that cooking improved in vitro protein digestibility of raw wingedbean, which was relatively low to that of other pulses (Table 1). Similar findings were reported earlier [56]. Jaya & Venkataraman [114] observed that germination improved

Table 1. Per cent Digestibility in vitro.

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Raw seed Cooked seed

Psophocarpus tetragonolobus 68,9 79.8 (Wingedbean var. USP-47) Glyeine max. (Soybean) 71, I 85.5 Phaseolus vulgaris 74.3 80.3 (var. castilla) Phaseolus vulgaris 75,2 83.7 (var. canario) Lupinus mutabilis 80,3 85.3 (Tarwi)

Source: Gross [79].

the in vitro carbohydrate digestibility of bengalgram and greengram but the findings of E1Faki et al. [57] showed that germination did not cause an increase in the in vitro carbohydrate digestibility of chickpea, horsegram and cowpea.

Saponins

Saponins occur in a wide variety of food plants [68, 177, 261]; bengalgram, soyabeans, navybeans, haricotbeans and kidneybeans being relatively rich. These seeds contained 56, 43, 21, 19 and 16g/Kg material, respectively (Table 2). The earlier reported values of saponin in soybean were quite low ranging between 0.46 to 0.50 per cent [21] showing wide differences. Recently, Sodipo & Arinze [234] have also reported low values of saponins in beans (Phaseolus vulgaris) containing 245.0 mg per Kg dry material. It is difficult to explain the

Table 2. Saponin content of legumes.

Seeds Saponin content (g/Kg dry material)

1. Bengalgram 56.0 2. Soybean 43.0 3. Navybeans Australia 21.0 Canada 6,7 USA 4.5

4. Greenbeans I3.0 5. Red Kidneybeans I6.0 6. Greengram 5.7 7. Lentil 3.7~4.6 8. Fababeans (Viciafaba) 4.3 9. Broadbeans 3.5

10. Greenpeas 11.0 11. Limabeans 1.1 12. Haricotbeans 19.0

Source: Fenwick and Oakenfull [68].

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reasons for the high values obtained by Fenwick & Oakenfull [68]. The presence of saponins in potatoes when eaten in large quantity causes abdominal pain, vomiting and diarrhoea. Soya products- tofu, textured vegetable protein and protein isolate are found to be good sources of saponins [67]. Saponins are also important in the human diet as they lower plasma cholesterol level in the animals and reduce the risk of heart diseases [179, 190-191]. It has also been reported [8] that they cause resistance among legume seeds against insect attack.

Saponins are not destroyed during cooking or processing. Raw navybeans or canned baked beans or canned broadbeans did not exhibit differences in the amount of saponins [21]. Fermentation was reported to reduce their level. Fermented soya product-tempeh, contained half the amount from that of raw soybean [190].

Proteolytic inhibitors

Protease inhibitors having ability to inhibit proteolytic activity of certain enzymes are found throughout the plant kingdom particularly among food legumes [134, 136, 146, 147, 213, 232, 242]. It has been demonstrated that the presence of these inhibitors in raw or dry-heat processed food legumes cause depression in growth and alteration in the pancreatic function of rats [17, 134, 232, 242], but it has been shown that these are largely inactivated by moist-heat processing [145]. These inhibitors have attracted the attention of nutritionists for the possible role which these substances might play in determining the nutritive value of plant proteins. But there is still uncertainty regrding their exact nutritional significance since there does not seem to be a clear-cut correlation between their amount and the beneficial effect which heat has on their nutritional values. Kakade et al [120, 121] found no correlation between the trypsin inhibitor activity and protein efficiency ratio of different varieties of soybean. Sitren et al. [232] concluded that the levels of trypsin inhibitor and lectin in soybean and peanut flours did not correlate with their overall biological impact in feeding studies with the rat.

Trypsin inhibitors possessing growth inhibitory property have been identifed in all food legumes in varying degrees [3, 44, 55, 80, 82-84, 90, 91,104, 120-122, 130, 133, 134, 147, 202, 204, 220, 221,233,235-237, 239, 245, 246, 258,259, 264]. These inhibitors have been isolated and characterised from different legume seeds [146, 147]. These include soybean [19], limabean [20], doublebean and fieldbean [206, 235, 237], navybean [263], adzukibean [268], bengatgram [14], black eyed pea [73], wingedbean [32, 100, 130-132, 222, 249, 250], kintokibean [250], guar seed [126] and blackgram [123]. These seeds contain one or more inhibitors having a molecular weight of about 8,000-10,000 [213], but soybean inhibitor has a molecular weight of 19,900. Soybean inhibitor has a unique property of combining with trypsin to form an inactive complex and thereby inhibit proteolytic activity. A comparative study on inhibitors from different legumes [192] revealed that soybean, fieldbean, kidneybean and bengalgram

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were more active in inhibiting the bovine pancreatic proteinases, whereas cowpea and redgram were more effective in inhibiting the human chymotrypsins. Trypsin inhibitor isolated from blackgram [123] has a molecular weight of 12,500. It has been characterised as non-competitive and stable at pH 2-10 within a temperature range of 30-80 °C for 30 minutes. It was identified as a glycoprotein, comprising of about 18.2 per cent carbohydrate and its one molecule containing 75 amino acid residues. Wingedbean contained two major trypsin inhibitors [13] having a molecular weight of about 20,000 and four cystine residues/molecule similar to soybean trypsin inhibitor. They have been characterised to be homogenous proteins. Inhibitor from wingedbean is specific for chymotrypsin. It inhibits bovine-chymotrypsin and not bovine trypsin. It has a molecular weight of 20,900 and contains four half-cystine residues/- molecule [131]. It is stable over the pH range 2.0-11.5 and to heating upto 70 °C at pH 4.1 and 8.0. Kintokibeans are found to contain at least five trypsin inhibitors having identical properties under gel filtration and ion-exchange chromatography but they differed among themselves in their specificity having two distinct sites of their activity towards trypsin and chymotrypsin [259]. Three of them resembled that of limabeans [95], kidneybeans [196] and garden beans [266] in respect of amino acid composition having higher amount of aspartic acid, serine and half cystine residues and containing no tryptophan. Studies by Tan-Wilson et al. [252] on Bowman-Birk proteinase inhibitors revealed that eight soybean strains had four to seven isoinhibitors species in each strain. Kaur & Bhatia [126] isolated three protein fractions from guar seed by Sephadex G-200 gel chromatography and found that all the three had considerable trypsin inhibitor activity. Hafez & Mohamed [90, 91] successfully separated non-protein trypsin and chymotrypsin inhibitors from soybean containing active peptides. They found that the inhibition of chymotrypsin is competitive and that of trypsin is non-competitive.

Different studies do not support the view that these inhibitors help protein breakdown during germination or prevent their degradation at seed maturation. It has been found that these inhibitors isolated from soybean [18] or pea [102] do not inhibit endogenous proteins of the same plant. Palmer et al. [185] reported an increase in the trypsin inhibitor activity in kidney-beans during germination but Collins and Sanders [40] and Noor et al. [176] did not observe any appreciable change in the activity in soybean or greengram, whereas others [58, 84, 126, 212] found reduced activity in greengram, blackgram, beans and guar seed. Trypsin and chymotrypsin inhibitor activities in beans were reduced by 62.9 and 67.1 per cent, respectively after five days of germination [212], while in case of guar seed, the trypsin inhibitor activity was reduced by 30 per cent after three days of germination [ 126]. Subbalakshmi et al. [243] reported similar findings on the changes in the trypsin inhibitor activity due to germination. Hobday et al. [102] and Baumgarten & Chrispeels [13] found that the protease inhibitor in pea or greengram is located outside the protein bodies.

Trypsin inhibitors are greatly affected by heat, genetic variance and stage of seed development [86]. Anti-tryptic activity among different food legumes

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Table 3. Trypsin and chymotrypsin activity in different legumes.

Seeds Trypsin inhibition (Units/mg protein)

Chymotrypsin inhibition (units/mg protein)

t. Hyacinth 16.97 17.59 (Dolichos lablab L.)

2. Lathyrus 11.88 11.25 3. Soybean 38.57 6.57 4. Cowpea 19.00 7.20 5. Bengalgram 8.57 2.79 6. Redgram 7.77 1.87 7. Blackgram 7.56 1.63 8. Greengram 5.36 - - 9. Clusterbean 2.45 0.07

Source: Sumanthi and Pattabiraman [245].

(Table 3) showed that beans were optimum; greengram being the lowest [103, 245]. Studies on soybean [124] have revealed that cotyledon component contained about 83 per cent of the total inhibitor activity of the seed while there was no activity in the embryo portion. Stage of soybean seed formation had a pronounced effect on its activity which gradually increased from early-milky to mature stage (Table 4) [124]. Soybean plant did not contain any activity at any stage of its growth. The rate of synthesis of the inhibitor was higher at the initial stages of seed formation. Phosphorus fertilization did not seem to influence its activity. It was concluded that the trypsin inhibitor was synthesised in the seed with initiation of seed formation and it was not translocated from the plant parts to the seed. Collins & Sanders [40] and Kute et al. [135] reported similar findings. They observed an increase in the trypsin inhibitor activity in soybean and wingedbean as the seed developed maturity.

These inhibitors in different legume seeds are inactivated by heat [3, 59, 124, 142, 146, 147, 183, 202, 220, 255]. Their inactivation is reported to greatly

Table 4. Trypsin inhibitor activity during developing soyben seed (expressed as per cent inhibition/ mg protein).

Variety Level of P Activity at different stages of seed Kg P2 05/ha development

Early Mid- milky milky

Mature

0 20.9 28.6 45.6 Bragg 56 20.9 28.9 45.7

112 20.6 29.0 45.6

0 2 t .4 27.9 46.0 Punjab-I 56 21.7 28.5 46.3

112 21.9 28.3 46,5

Source: Kapoor and Gupta [124].

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improve the nutritive value of their seed proteins [22, 55, 122, 141,202, 233]. The presence of residual tannins in the cooked beans accounted for the inhibitor activity remaining after heat treatment, which was hardly one per cent of the total activity [44, 60, 240, 248]. Temperature, duration of heating, particle size and moisture are found most beneficial in destroying the inhibitor activity [3, 147, 202, 204]. Antunes & Sgarbieri [7] reported that this inhibitor was a heat-resistant antinutritional constituent. A high initial moisture content (20 per cent) in soybean favoured rapid destruction of the inhibitor [3, 99]. Sitren et al. [232] found that raw soybean caused the maximum (76 per cent) inhibition of the trypsin enzyme, the dry-heated soybean caused 61 per cent inhibition, and the moist-heated soybean caused the least inhibition as 11 per cent. Studies by Kapoor & Gupta [124] showed that autoclaving of soybean seed for 30 minutes or steaming for 60 minutes completely inactivated the inhibitor activity. How- ever, soaking for 8 hours prior to steaming considerably reduced its time of inactivation to 15 minutes. The inhibitor was completely inactivated by direct injection of steam for two minutes followed by cooking for 20 minutes at 4.2 Kg/sg.cm. pressure [220]. Studies on the kinetics of the inhibitor activity in relation to temperature and moisture [189] showed that the loss of activity was of first order reaction. The first order rate constants for loss of activity in cowpea flour containing 7.5, 19.4 and 26.5 per cent moisture when heated at 100 °, 125 ° and 150°C for periods of 0.5 to 120 minutes ranged from 1 x 10 -2 to 18 rain -l . Buere et al. [28] also reported that the trypsin inhibitor activity loss in beans by temperature followed first order reaction kinetics and the rate of loss greatly increased with increasing moisture from 2-55 per cent. Dielectric heating [24, 230], infra-red cooking [66] and microwave radiation [267] were also reported to inactivate the inhibitor, causing improvement in the nutritional quality of soybean. Ellendrieder et al. [61] reported the presence of proteinous substances which accelerated thermal inactivation of trypsin inhibitor. There was a loss of 90 per cent in the trypsin inhibitor activity in the crude extract from kintokibeans at 100°C for 60 minutes but it was observed that the purified trypsin inhibitor was heat-stable and was inactivated by heat in the presence of high molecular weight proteins [258]. Doell et al. [49] found that most of the inhibitor activity was lost during processing of soya foods and the remaining was eliminated during cooking. It has also been found that soaking in one per cent salt solution and sodium bicarbonate has considerably reduced the inhibitor activity of common Indian pulses [239].

Different varieties of soybean are found to differ in their trypsin inhibitor activity [80]. Singh & Eggum [231] reported large variation in the levels of protease inhibitors of pigeonpea varieties. The concentration of these inhibitors was higher in some of its wild species. Trypsin and chymotrypsin inhibitor activity in wingedbean varied from 22.2 to 42.5 mg of trypsin inhibited and from 30.1 to 47.6mg of chymotrypsin inhibited per g of seed material, respectively [133]. The inhibitor activity in the seed was also shown to be influenced by location [82].

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Lathyrogens

In India and other countries, people consuming lathyrus (Lathyrus sativus) for prolonged periods suffer from a paralytic disease known as lathyrism, which causes a public health hazard. The incidence of lathyrism in areas where lathyrus used to be consumed has been established. Various aspects of this problem have been reviewed from time to time [85, 89, 94, 146, 219].

Human lathyrism in India is widely common where the crippling disease afflicts the poorer sections of the people especially under conditions of famine resulting from droughts when the field crops miserably fail, and lathyrus forms the staple diet because this pulse is highly drought resistant, and as an alternate crop, it is cultivated on lands subjected to drought, excessive rain or floods. In India, this pulse occupies nearly 5 million acres under cultivation, which is 4 per cent of the total area under pulse crop and constitutes 3 per cent of the total pulse production; Madhya Pradesh produces more than 50 per cent of the total produce in the country [89]. The various reports have confirmed that prolonged consumption of this pulse for a period of three to six months afflicts the central nervous system characterised by weakness and paralysis of the leg muscles and death in extreme cases (neurolathyrism). This disease affects mostly young men between 20 and 29 years of age. A 1958 epidemiological survey in India indicated that in a single district of Rewa in Madhya Pradesh, there were as many as 25,000 cases of neurolathyrism in a total population of 634,000 [53]. Another survey in 1974 in Raipur (Madhya Pradesh) showed that the rate of prevalence of lathyrism was 40 per 1,000 [54]. The number of cases were over 1,00,000 in 1975 [175]. Studies by Attal et al. [11] revealed that there were typical differences and characteristics features in cases of lathyrism in Amgaon area (Distt. Bhandara) of Maharashtra.

Various attempts have been made to identify the causative agent of human lathyrism. The presence of certain phenolic compounds, a toxic alkaloid, a water soluble toxic amine, excess quantity of manganese, an alkaline volatile liquid, presence of selenium which interferes in methionine metabolism and presence of pathogenic fungus which grows,on the pulse during wet and virus infection have been attributed by different workers to be causative factors for the toxicity [174, 244]. But none appeared to be conclusive.

Several groups of workers in India [1, 170, 215] isolated a neurotoxic compound from the lathyrus seeds, and characterised it as fl(N)-L-e, fl-diaminopropionic acid (BOAA or ODAP) to ~oe the causative principle of human neurolathyrism. Its injection in young chicks, rats and monkeys was shown to produce severe neurotoxic symptoms [184, 215]~ The effect of BOAA in experimental animals made its involvement in human lathyrism highly suggestive. However, its oral feeding as well as parenteral administration of the seed extracts to experimental animals even for prolonged periods did not induce paralysis [173]. Monkeys fed on the pulse as the sole diet source for almost one year did not show any external symptoms of neurological behaviour.

In addition to BOAA, another water soluble aliphatic amino acid glycoside

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with a nitrile group was isolated from the lathyrus seeds and shown to be toxic to day-old chicks at a dose of 50mg per 100g body weight. This compound produced paralysis of the limbs within 5 to 10 minutes of intraperitoneal injection [216, 2t7]. It was designated as N-fl-D-glucopyranosyl-N-a-L- arbinosyl-e, fl-diaminopropionitrile. This compound was reported to act syner- gistically with BOAA.

The neurotoxic compound (BOAA) is reported to undergo transamination in rat tissue giving to a keto acid product [36], which inhibits the growth of several micro-organisms. It has been found [215] that a 30 per cent ethanol extract of lathyrus meal caused neurological symptoms in day-old chicks, and intraperitoneal administration of BOAA in two days-old rats caused typical convulsions within 10 minutes [173]. It was concluded [38] that BOAA interfered with the ammonia generation of fixing mechanism in the brain and led to chronic ammonia toxicity. Biochemical studies [35, 37] with brain homogenates prepared from the BOAA injected rats revealed an increase in transglutaminase, protease, glutaminase, adenosine deaminase and transaminase levels, and a decrease in the brain glucose, glycogen, ATP, phosphocreatinine and acetylch- oline levels of the convulsing animals. It was concluded that protein bound and free amide group could be the source of ammonia in BOAA treated rats, and BOAA was a typical convulsant.

Earlier studies [209] on BOAA administered through the lumbar route in monkeys supported the concept of a blood brain barrier but the later work [207] did not support the concept of effective blood brain barrier to the toxin. The existence of a blood brain barrier in human beings is yet to be established.

It was shown [208] that treatment of adult animals with drugs or chemicals known to induce an acidotic condition would make them susceptible to the toxin. It was interpreted that acidosis had favoured increased entry of the toxin into the brain. BOAA was detected in the brain of young rats and acidotic rats but not in adult rats [34].

Recent studies on the effect of BOAA on glutamate metabolism revealed [137] that this compound could be a potential antagonist of glutamate. These investigations established that BOAA was neurologic and the amino group was involved in causing neurotoxic symptoms.

Studies on the biosynthesis of this neurotoxin compound [154] revealed that it was synthesised by the action of oxalyl-COA synthetase and oxalyl-L-e, fl-diamino-propionic acid oxalyl transferase enzymes. Neurotoxin was synthesised by the oxalylation of L-e, fl-diaminopropionic acid [155] as follows:

Mg++ Oxalate + ATP + COA . " Oxalyl-COA + AMP + PPi (i)

Oxalyl-COA- synthetase

Oxalyl-COA + L-e, fl-diaminopropionic acid ~ BOAA + COA synthetase

(ii)

A partial resolution of these two enzymes activities using C.M. Sephadex column was reported [156]. Studies on the factors involved in the manifestation

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of the paralytic disease and their mode of action, however, need further inves- tigation.

There has been no effective medicine for the cure of lathyrism and the disease could only be checked by feeding a nutritive diet to the affected person and by discontinuing the consumption of this crippling pulse. A possibility of some improvement by indigenous drugs was reported by Attal et al. [11] but it is lacking confirmation.

This pulse wildly grows in a number of countries including India, and con- tinues to cripple a large section of the poor people. Therefore, certain procedures have been evolved to identify its adulteration among pulses employ- ing visual, microscopic, chemical and chromatographic tests [48, 52, 93, 172]. Also simple domestic procedures for detoxification have been proposed [166]. These include steeping the dehusked seeds overnight followed by steaming for 30 minutes and subsequently rejecting the extract. In this process, many of the water soluble nutrients are lost, and need supplementation. Roasting of the seed at 140 °C for 15-20 minutes has also been advocated. Ramachand et al. [205] suggested that the fermentation method with the use of bacillus species may be exploited as they observed that neurotoxin is broken down by this bacillus without affecting the nutritional quality of the seed.

The utility of such domestic procedures could not be adopted on a national scale because of certain practical difficulties. As an alternate approach, some attempts are being made to evolve genetically new strains of lathyrus to be low or free from toxin. Analysis of a large collection of lathyrus seeds revealed a wide variation in the amount of BOAA ranging from 0.1 to 2.5 per cent [171, 238]. These seeds have also been found to contain other compounds which could be lathyrogenic. These compounds were identified to be e, 7-diaminobutyric acid and/?-cyanoalanine producing neurotoxic effects when injected into animals. Thus, the role of these compounds have to betaken into consideration in the pathogenesis of lathyrism for achieving a breeding programme successfully.

In India, this pulse is otherwise a boon to the poor people as an article of diet especially in times of famine when lathyrus is the only available food in certain regions of the country particularly as it is rich in proteins and lysine, and is potentially useful as a protein and lysine supplement. Therefore, it is necessary to take preventive steps to minimise or to eliminate the incidence of human lathyrism.

Phytohemagglutinins

Substances possessing the property to agglutinate red blood cells are known as phytohemagglutinins (lectins). These substances, protein in nature, are widely distributed among food legumes [76, 106, 144, 146, 149, 255]. Except concanavalin A lectin from jackbean, these lectins in food legumes are glycopro- teins containing 4 to 10 per cent carbohydrates. They constitute about 2 to 10 per cent of total protein in most leguminous seeds. Besides their ability to

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agglutinate red blood cells, Liener [146] has reported that they exhibit other chemical and biological properties, which include interaction with specific blood groups, mitogenesis, agglutination of tumor cells and toxicity towards animals.

Various microbiological and immunochemical studies [106, 129, 199, 201, 265] have provided evidence that the higher intake of lectins produce toxic effects, inhibiting growth and causing death. It is believed that these lectins bind intestinal mucosal cells, causing malfunction, disruption and lesion in the small intestine and thereby interfere with the absorption of nutrients from the gut [106, 143,201,253]. Pusztai et al. [200] have reported that a proportion of bound lectin goes to the circulating system. Others [64, 149] have also supported the view that binding of lectins to the cells lining the intestine may cause a serious impairment in the ability of the cells to absorb nutrients from the gastrointesti- nal tract and thus inhibiting growth and in extreme cases, death. Gatehouse et al. [72] have suggested a similar mechanism of lectin toxicity in insects that the ingested lectin causes disruption of the epithelical cells of the larvae midget leading to breakdown of the transport of nutrients into these cells, and the absorption of potentially harmful susbtances. Some others [12, 115, 116] have advocated that bacteria also play an important role in causing toxicity by lectins. Others [116(a), 211] indicated that these lectins interfere with the digestive enzymes and reduce the availability of nutrients. Studies by Thompson et al. [253] have shown that the addition of lectins at the same concentration present in raw bean extract, decreased the rate of digestion of heat-treated bean extract, casein and bovine serum albumin to levels close to that of raw bean extract. They concluded that lectins affected the activity of digestive enzymes. It was recently shown [78] that diets containing kidneybean lectins severely disrupted the brush borders of duodenal and jejunal enterocytes of rats aged between 30 and 123 days, and that oral immunisation did not protect the rats from the effects of toxicity, and that the immune response was a result of continuous absorption of lectin throughout the feeding period. The extent of toxicity was not affected by the age of the animals.

Their nutritional significance and physiological functions were earlier reviewed [143, 144]. In the biological system, they have a unique property of binding saccharides and saccharides-containing proteins. It was speculated [254, 255] that they (a) act as antibodies to counteract soil bacteria; (b) serve to protect plants against fungal attack by inhibiting fungal polysaccharides; (c) serve to transport or store sugars; (d) act as glue for attaching glycoprotein enzymes in organised multi-enzyme systems; and (e) play a key role in the development and differentiation of embryonic cells. It has also been proposed that they are involved in the symbiotic relationship between leguminous plants and bacteria in binding to the root nodules of the plant and in the interaction of legumes with bacteria in the nitrogen fixing system as they were detected in the roots of Phaseolus vulgaris [92], and were also found to be associated with bacteria nodulating soybean [23]. It has been shown that kidneybean lectins protect plants against insect [72, 113], fungal [165] and pathogenic bacterial attack [227]. Gatehouse et al. [72] have shown that both albumin and globulin protein

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fractions from kidneybean containing hemagglutinating activity were toxic when ingested by the developing larvae of the bruchid beetle (Caltosobruchus maculatus), implicating the seed lectins as being involved in seed resistance. Jayne-Williams & Burgess [116] have postulated that binding of lectins to the cells lining the intestine may interfere with the normal defence mechanism of the cells.

There are over 600 species of leguminosae family which exhibit hemagglutinating activity [255]. These lectins are found to be localised in the cytoplasm of the cotyledon and embryonic cells, and greatly reduce on germination. These have been isolated and characterised from different sources which include Phaseolus species--kidneybean, navybean, limabean, waxbean, etc. [6, 27, 46, 77, 107, 110, 111,158, 159, 197, 226, 229, 247]; lentil [69, 76, 117, 241]; pea [29, 30, 62, 63, 160, 186, 256, 257, 262]; horsegram [158, 187]; soybean [151]; fieldbean [206, 218]; broadbean [158]; j ackbean [182] and wingedbean [74, 108, 133, 195, 251].

Kidneybean lectins are of considerable nutritional significance as they constitute an important source of dietary protein in several parts of the world but they are toxic to both mammals and birds when ingested [65, 116, 198] and are poorly digested by rats [129, 199]. Lectins from this source are found to differ in their specificity depending on animal species, human blood group and pre- treatment of cells with proteolytic enzymes [27, 107, 110, 229]. The presence of five heterogenous proteins in kidneybeans [164] each consisting of isomeric noncovalently bound tetramere is reported to be responsible for differences in the lectin activity in its different species. Others [27, 111, 149] identified two groups to distinguish specificity of lectin activity. One isolated from P. vulgaris was non-specifc with human erythrocytes of all blood groups and the other isolated from P. lunatus was specific for A blood group cells. Wingedbean hemagglutinins showed strong reaction with human and rabbit bloods and caused higher toxicity [74, 108, 133, 195] than that of soybean [79]. Gillespie et al. [74] identified two distinct groups of wingedbean hemagglutinins (isolectins) differing in specificity towards erythrocytes. Kortt & Cardwell [133(a)] isolated acidic and basic lectins from six varieties of wingedbeans collected from different regions of south-east Asia, and found no difference in their properties, particularly in respect of their amino acid composition. These isolated lectins slightly differed in their specificity. The acidic lectins agglutinated trypsinised human type A, B and O erythrocytes but not trypsinised rabbit erythrocytes, whereas the basic lectins did not agglutinate trypsinised human type O erythrocytes but were able to agglutinate trypsinised rabbit and human type A and B erythrocyte. There was little varietal differences in the wingedbean hemagglutinin activity [133]. Its activity varied from 40 to 320 units per mg of wingedbean [251]. Rasanen et al. [210] obtained a leuco-agglutinin from kidneybean in a crystaltised form having a molecular weight of 126,000 consisiting of four identical units. Other lectins isolated from waxbean, navybean, limabean, lentil, soybean and horsegram had a molecular weight of 30,000, 32,000, 124,000, 52,000, 122,000 and 120,000, respectively [6, 69, 77, 151, 187, 226].

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There were two principal lectins in pea [63, 160] and the third one which was identified, was due to hybridization of the other two [62]. They were found to require Ca + + and Mn + + for their activity [186] and a tryptophan residue played an important role [29]. Pea also contained two mitogenic lectins having similar properties to the other ones [256]. Pea lectins [257] were similar to that of soybean [138] and lentil [241] having two binding sites for mannose and methyl ct-D-glucoside.

Their growth inhibitory property in beans is well recognised [106, 146]. Lectin isolated from soybean was shown to inhibit growth of rats [140]. It accounted for 25 per cent of the growth inhibition which raw soybean produced in rats. On the other hand, feeding trials with soybean [260] showed that its lectin had little direct effect on the nutritive properties of its protein. Soybean lectin was first isolated by Liener [139], and its characteristics were studied by Sharon and his co-workers [150, 151 ]. Fieldbean tectin had a similar growth inhibiting effect on rats. It caused zonal necrosis of the liver [206, 218]. Bhatty & Christison [17] obtained similar findings with fababean and lentil. They concluded that lectins were responsible for the poor growth obtained with fababean and lentil meals, concentrates and isolates.

There were more than 200 Phaseolus species which exhibit hemagglutinating activity in varying degrees. The larger group, represented by Phaseolus vulgaris, reacts nonspecifically with human erythrocytes of all blood groups and the smaller group, represented by Phaseolus lunatus, is specific for A blood group cells [144]. Kidneybean and blackbean were found to display a significant level of hemagglutinin activity of 3560 and 2450units/ml (Table 5) [103]. They inhibited growth at levels as low as 0.5 per cent of the diet; kidneybean lectin being more effective than the blackbean lectin. Kidneybean lectin caused 100 per cent mortality at 0.5 per cent level which blackbean lectin produced a similar mortality rate at 1.2 per cent level. A high level of hemagglutinin in kidneybeans

Table 5. Hemagglutinating and antitryptic activities of crude extracts* of raw legumes.

Legume Hemagglutinating activity (units/ml)

Antitryptic activity (units/ml)

Phaseolus vulgaris Black bean 2450 2050 Kidney bean 3560 1552

Cicer arietinum 0 220 (Bengalgram) Cajanus eajan 0 418 (Redgram) Phaseolus aureus 0 260 (Greengram)

* A 10 per cent suspension of the finely ground meal in one per cent NaC1, clarified by centrifuga- tion. Source: Honavar et al. [103].

214

Table 6. Effect of heat treatment* on the hemagglutinating activity in different legumes.

Species Common Hemagglutinin (units/g) name

Raw Heat treated

Phaseolus Natal round 1,55,000 50 vulgaris yellow bean Phaseolus Umzumbi bean 45,000 0 vulgai"is P. vulgaris Haricotbeans 40,000 20 Glyeine max. Soybean 30,000 0 Vicia sativa Common vetch 120 10 Vicia faba Broadbean 90 11 Pisum sativum Green pea 80 4 Phaseolus aureus Mungbean 78 0 Vigna sinensis Cowpea 6 2

* Heat treatment---autoclaving for 30 minutes at 121 °C. Source: DeMuelenaere [46].

(over 10 per cent of total protein) was also found by Pusztai et al. [199]. Yellow bean, umzumbibean, haricotbean and soybean contained very high hemagglutinin; their activity being 155,000, 45,000, 40,000 and 30,000 units per gm, respectively [46], whereas Vicia sativa, broadbean, greenpea, cowpea, mungbean, bengalgram and redgram were relatively either very low or almost free (Tables 5 and 6). Lectins from limabean, bengalgram, greengram, redgram and castorbean were non-toxic [46, 103, 158, 182]. However, Manage et al. [158] observed that oral administration of limabean lectin had a depressing effect on the growth of rats.

These lectins resist inactivation by dry heat [45, 251]. Moist-heat treatment however eliminates the toxicity [109]. Similar findings were obtained by Sitren et al. [232]. They observed that raw and dry-heated soybean contained 112.1 ktg/ g and 94.7/zg/g lectin, respectively, whereas moist-heated soybean was completely free of lectin. Autoclaving for a shorter period (5-30 minutes) completely destroyed the bean activity [45, 46, 118, 133, 251] (Table 6). ManciniFilho et al. [t 59] reported that ionizing radiations with a dose of 50 Krad destroyed 50 per cent of the activity.

Favism

Favism is a disease characterised by hemolytic anaemia affecting certain individuals consuming raw or cooked broadbeans (Viciafaba) [15, 146, 155]. The disease is common among the inhabitants of the Middle East, North Africa, Italy and Greece [15]. The blood cells of the affected individuals exhibit a number of biochemical abnormalities, the most significant of which are a diminished level of reduced glutathione and glucose-6-phosphate dehyd-

215

rogenase activity. These beans contain anti-nutritional compounds which are thermolabile [31, 161] and thermostable [179, 180].

Various studies [16, 39, 43, 148, 152, 153, 168, 169, 179, 181] have implicated vicine and convicine occurring naturally as glucosides and their respective aglycons divicine and isouramil pyrimidines as the causative agents of favism. It has been reported [70, 97] that these aglycones are produced in the large intestines on consumption of fababeans, and are transported to the blood, where they form certain products in the presence of oxygen [2, 39], causing favism in sensitive individuals. Muduuli et al. [168] demonstrated that these compounds elevated the levels of plasma lipid and lipid peroxides, liver peroxides and glutathione and erythrocyte hemolysis in vitro and depressed plasma vitamin E levels. In vitro studies revealed that divicine and isouramil pyrimidines caused a rapid decrease in the glutathione content of glucose-6- phosphate dehydrogenase deficient red blood cells, an effect which was accounted for the hemolytic activity exerted by broadbeans [142]. Divicine pyrimidine was also found to be toxic when injected into experimental animals, and because of its occurrence in Vicia sativa, it was, at one time, considered as the causative factor of lathyrism. Divicine compound as the causative factor of favism is not final as its occurrence is not confined to broadbeans.

The concentration of vicine and convicine compounds in raw fababeans has been found to be 5.3 mg and 2.3 mg per gm material, respectively [9]. A new method for quantitation of these compounds was developed [38(a)]. These compounds were earlier isolated and their properties were studied [163]. Arbid & Marquardt [10] have developed a procedure for the purification of these compounds while a rapid reverse-phase liquid chromatography method was developed for their determination [162].

Certain workers [26, 98, 112] have suggested that it may be possible to screen fababean cultivars to effectively eliminate these compounds because except for the toxic principle, these beans are of good nutritional quality [50, 51]. A number of procedures have been proposed to reduce or eliminate these compounds. These include development of cultivars free from these compounds, selective destruction of these compounds, and extraction of the compounds from the seeds. Gardiner et al. [71] screened a number of fababean genotypes having variation in the concentration of these compounds, but no strain, free of these compounds, has been identified. Hegazy & Marquardt [96] succeeded in extracting these compounds completely, but the seeds after extraction were rendered unpalatable. Mager et al. [153] have earlier demonstrated that these compounds could be easily hydrolysed by/%glucosidase enzyme from almonds. Recent studies by Arbid & Marquardt [t9] have shown that 88 to 89 per cent of these compounds were hydrolysed when one gram of fababean paste was mixed with 0.1 gm of almond powder and 0.1 ml of lemon juice, and incubated at 30 °C for three hours. Their studies have revealed that the concentration of these compounds in the fababean food preparation is greatly reduced by enzym- ic hydrolysis.

216

Table 7. Cyanide content of certain plants.

Plant HCN yield (rag/100 g)

1. Limabean (Phaseolus lunatus) 210.~312.0 Samples incriminated in fatal human poisoning Normal levels 14.4-16.7

2. Sorghum 250.0 3. Cassava 113.0 4. Linseed meal 53.0 5. Black-eyed pea (Vigna sinensis) 2.1 6. Garden pea (Pisum sativum) 2.3 7. Kidneybean (Phaseolus vulgaris) 2.0 8. Bengalgram (Cicer arietinum) 0.8 9. Redgram (Cajanus cajan) 0.5

Source: Liener [146] and Montgomery 1167].

Cyanogenetic glycosides

Legumes also exhibit toxicity because of their cyanide producing potential. They contain certain cyanogenetic glycosides which release HCN on hydrolysis [41, 42, 142, 146, 167, 225]. Cassava and limabeans predominantly contain these glycosides. Limabeans (Phaseolus lunatus) were reported to be responsible for serious outbreak of human poisoning and cases of human intoxication. The yield of HCN from limabeans varied from 210.0 to 312.0mg/100g (Table 7). Such a high level of HCN has been the causative factor for fatal human poisoning. However, the yield of HCN from bengalgram, redgram, peas and kidneybean is negligible ranging from 0.5 to 2.3 mg/100 g, and therefore do not exhibit toxicity. A recent report [107] showed that soy protein products and soybean hulls contained 0.07-0.3 and 1.24/~g/g of HCN, respectively, whereas in the earlier report [125], soybean was shown to be free of cyanogenetic compounds. HCN in cereal grains and products is negligible ranging from 0.001 to 0.45 #g/gm [41] and is of little nutritional significance.

It has been shown that most of the liberated HCN is lost by volatilization during cooking of the limabean, and cyanide is rapidly converted to thiocyana- tes or other compounds, but cases of human intoxication and chronic neurological effects occurred even with cooked limabeans [167].

Conclusion

Some other substaflces like toxic amino acids, goiterogens, anti-vitamin factors, etc., which also interfere and cause toxicity, have not been covered. The present write-up indicates that these substances are widely common in diferent food legumes consumed by human beings, but their ill-effects are not often manifest,

217

suggesting thereby that foods producing immediate ill-effects are either avoided or ways and means have been devised to eliminate them. For instance, cooking itself and other common means of preparation have proved effective in destroying many of the toxic constituents. However, in some cases, complete detoxification does not take place. Inadequate commercial processing of soybean products is an example. The incidence of lathyrism from consumption of lathyrus in times of famine, and development of favism disease from consuming broadbeans are common.

References

1. Adiga PR, Padmanabhan G, Rao SLN, Sarma PS (1962) The isolation of a toxic principle from Lathyrus sativus seeds. J Sci Industr Res 21C: 284-286

2. Albano E, Tomasi A, Mannuzzu L, Arese, P (1984) Detection of a free radical intermediate from divicine of Viciafaba. Biochem Pharmacol 33:1701-1704

3. Albrecht WJ, Mustakes GC, McGhee JE (1966) Rate studies on atmospheric steaming and immersion cooking of soybean. Cereal Chem 43:400-407

4. Ambe KS, Sohonie K (1956) Studies on trypsin inhibitors in Indian foodstuffs. Part II. Trypsin inhibitor in germinating doublebean and fieldbean, and their growing plants. J Sci Industr Res 15C: 136-140

*5. Anderson RL, Rackis JJ, Tallent WH (1979) In: Wilcke HL, Hopkins DT, Waggle DH (eds) Soy Protein and Human Nutrition pp 209 233. New York: Academic Press

6. Andrews AT (1974) Navy (Haricot) bean (Phaseolus vulgaris) lectin isolation and characterisation of two components from a toxic agglutinating extract. Biochem J 139:421~429

7. Antunes PL, Sgarbieri VC (1980) Effect of heat treatment on the toxicity and nutritive value of dry bean (Phaseolus vulgaris) protein. J Agric Fd Chem 28: 935-938.

8. Applebaum SW, Marco S, Birk Y (1969) Saponins as possible factors of resistance of legume seeds to the attack of insects. J Agric Fd Chem 17:618~i22

9. Arbid MSS, Marquardt RR (1985) Hydrolysis of the toxic constituents (vicine and convicine) in fababean (Vicia faba L.) food preparations following treatment with /~-glucosidase. J Sci Fd Agr 36:839-846

10. Arbid M SS, Marquardt RR (I 985) A modified procedure for the purification of vicine and convicine from fababeans (Viciafaba L.). J Sci Fd Agr 36:1266-1270

11. Attal HC, Kulkarni SW, Choubey BS, Palkar ND, Deotale PG (1978) A field study of lathyrism--some clinical aspects. Indian J Med Res 67:608415

* 12. Banwell JG, Boldt DH, Meyers J, Weber FL, Miller B, Howard R (1983) Phytohemaggluti- nin derived from red kidneybean (Phaseolus vulgaris): A cause for intestinal malabsorption associated with bacterial overgrowth in the rat. Gastroenterology 84: 506.

13. Baumgarten B, Chrispeels MJ (1976) Partial characterization of a protease inhibitor which inhibits the major endopeptidase in cotyledons of the navybean. Plant Physiol 58: 1 ~

14. Belew M, Porath J, Sundbert L (1975) The trypsin and chymotrypsin inhibitors in chickpeas (Citer arietinum L.)--purification and properties of the inhibitors. Eur J Biochem 60: 247-258

15. Belsey MZ (1973) The epidemiology of favism. Bull W.H.O. 48:1-13 16. Benatti U, Guida L, DeFlora A (1984) The interaction of divicine and glutathione and

pyrimidine nucleotides. Biochem Biophys Res Commun 120:747-753 17. Bhatty RS, Christison GI (1984) Composition and nutritional quality of pea (Pisum stivum

*Originals not available.

218

L.), fababean (Vicia culinaris Medik.) meals, protein concentrates and isolates. Qual Plant Plant Fds Hum Nutr 34:41-51

18. Birk Y (1968) Chemistry and nutritional significance of proteinase inhibitors from plant sources. Ann NY Acad Sci 146:388-399

I9. Birk Y (1976) Trypsin and ch}a-notrypsin inhibitors from soybeans. Methods Enzymol 45: 700707

20. Birk Y (1976) Limabean trypsin inhibitors. Methods Enzymol 45:707-709 21. Birk Y (1980) Saponins. In: Liener IE (ed.) Toxic Constituents of Plant Foodstuffs, 2nd edn.

pp 169-211. New York: Academic Press 22. Birk Y, Gertler A (1961) Effect of mild chemical and enzymatic treatment of soybean meal

and soybean trypsin inhibitors on their nutritional and biochemical properties. J Nutr 75: 379-387

23. Bohlool BB, Schmidt EL (I971) Lectins: A possible basis for specificity in the Rhizobium --legume root nodule symbiosis. Science, 185:269--271

24. Borchers R, Manage LD, Nelson SO, Stetsen L (1972) Rapid improvement in nutritional quality of soybeans by dielectric heating. J Fd Sci 37:333-334

25. Bradbear N, Boulter D (1984) The use of enzymatic hydrolysis in vitro to study the digestibility of some Phaseolus seed proteins. Qual Plant Plant Fds Hum Nutr 34:3-13

26. Brown EG, Roberts RM (1972) Formation of vicine and convicine by Viciafaba. Phytoch- em II: 32023206

27. Brucher O, Wecksler M, Levy A, Polozzo A, Jaffe WG (1969) Comparison of phytochemag- glutinin in wild beans (Phaseolus aborigineus) and in common beans (Phaseolus vulgaris) and their inheritance. Phytochem 8:1739-1743

28. Buera MP, Pilosof AMR, Bartholomai GB (1984) Kinetics of trypsin inhibitory activity loss in heated flour from bean, Phaseolus vulgaris. J Fd Sci 49:124-126

29. Bures L, Entlicher G, Kocourek J (1972) Studies on phytohemagglutinins. XI. Importance of tryptophan residues for the activity of pea phytohemagglutinin. Biochim Biophys Acta 285:235-242

30. Bures L, Entlicher G, Ticha M, Kocourek J (1973) Studies on phytohemagglutinins. XV. Hemagglutinins of the pea and lentil: Advantage of their application in comparison to concanavalin A in some agglutination reactions. Experientia, 29:1546-1547

31. Cansfield PE, Marquardt PR, Campbell LD (1980) Condensed proanthocyanidine of fababeans. J Sci Fd Agr 31:802-812

32. Chan J, deLumen BO (1982) Properties of trypsin inhibitor from wingedbean seed isolated by affinity chromatography. J Agric Fd Chem 30:4246

33. Chatterbuck KL, Kehrberg NL, Marable NL (1980) Solubility and in vitro digestibility of soy flours, concentrates and isolates. J Fd Sci 48:931-935

34. Cheema PS, Padmanabhan G, Sarma PS (1969) Neurotoxic action of/LN-oxalyl-L-c~, /~-diaminopropionic acid in acidotic adult rats. Indian J Biochem 6:146-147

35. Cheema PS, Padmanabhan G, Sarma PS (1970) Biochemical characteristion of/~-N-oxalyl- L- c~, /Ldiaminopropionic acid, Lathyrus sativus neurotoxin as an exitent amino acid. J Neurochem 17:1295-1298

36. Cheema PS, Padmanabhan G, Sarma PS (1971) Transamination of/~-N-oxalyl-L-c~,/~-diaminopropionic acid--Lathyrus sativus neurotoxin in tissues of rat. Indian J Biochem 8: 1619

37. Chemma PS, Padmanabhan G, Sarrna PS (1971) Mechanism of action of/~-N-oxalyl-L-c~, /Ldiaminopropionic acid, the Lathyrus sativus neurotoxin. J Neurochem 18:213%2144

38. Cheema PS, Padmanabhan G, Malathi K, Sarma PS (1969) The neurotoxicity of fl-N- oxalyl- L-a, /Ldiaminopropionic acid, the neurotoxin from the pulse Lathyrus sativus. Biochem J 112:29-33 Chevion M, Navol T (1983) A novel method for quantitation of favism involving agents in legumes. Anal Biochem 128:15~158 Chevion M, Navol T, Glaser G, Mager J (1982) The chemistry of favism-inducing corn-

38(a).

39.

219

pounds. The properties of isouramil and divicine and their reaction with glutathione. Eur J Biochem 127:405-409

40. Collins JL, Sanders GG (1976) Changes in trypsin inhibitor activity in some soybean varieties during maturation and germination. J Fd Sci 41:168-172

"41. Corm EE (I979) In: Neuberger A, Jukes TR (eds) Biochemistry of Nutrition, Vol. 27, pp 21-42. Baltimore, MD: University Park Press

*42. Conn EE (1981) In: Ayres JC, Kirschman JC (eds) Impact of Toxicology on Food Process- ing, pp 105-121, Westport, CT: Avi

43. D'Aquino M, Gaetani S, Spadoni MA (1983) Effect of factors of favism on the protein and lipid compounds of rat erythrocyte membrane. Biochim Biophys Acta 731:161-167

44. deLumen BO, Saiamat LA (1980) Trypsin inhibitor activity in wingedbean (Psophocarpus tetragonolobus) and the possible role of tannin. J Agric Fd Chem 28:533-536

45. DeMuetenaere HJH (1964) Effect of heat treatment on the hemagglutinating activity of legumes. Nature (Lond.), 201:1029-1030

46. DeMuelenaere HJH (1965) Toxicity and hemagglutinating activity of legumes. Nature (Lond.), 206:827-828

*47. Deshpande SS, Sathe SK, Cheryan M (1983) In vitro proteolytic digestibility of dry bean Phaseolus vulgaris L. protein concentrates. Nutr Rep Intl 27:931-936

48. Diwaker NG, Nagarajarao UN (I 972) A paper chromatographic method for the detection of Lathyrus sativus in presence of some common edible pulses using ultraviolet light. J Instn Chem (India) 44:163-165

49. Doell BH, Ebden CJ, Smith CA (1981) Trypsin inhibitor activity of conventional foods which are part of the British diet and some soya products. Qual Plant Plant Fds Hum Nutr 31:139-150

50. Duthrie IF, Porter PD, Gadaly B 91972). Nutritional value of a protein isolate from the Throws M.S. variety of fieldbeans (Viciafaba L.) grown in the U.K. Proc Nutr Soc 31: 1980-1981

51. Duthrie IF, Owen E, Miller, EL, Laws BM, Owers MJ (1977) Preparation of fieldbean (Vicia faba) cotyledons as a substitute for dried skim-milk in calf feeding. Proc Nutr Soc 35: A 1t5-116

52. Dutta AB (1965) A chemical method for the detection of khesari pulse (Lathyrus sativus). J Proc Instn Chem (India) 37:16

53. Dwivedi MP, Prasad BG (1964) An epidemiological study of lathyrism in the district of Rewa (Madhya Pradesh). Indian J Med Res 52:81-116

54. Dwivedi MP, Mishra SS (1975) Recent outbreak oflathyrism and experience with propaga- tion of detoxified Lathyrus sat#us. Proc Nutr Soc (India) 19:23-30

55. Egberg DC, Potter RH, Honald GR (1975) The semiautomated determination of trypsin inhibitors in textured soy protein. J Agric Fd Chem 23:603-605

*56. Ekpenyong TE, Borchere RL (1978) Nutritional aspects of wingedbean In: The Winged Bean. Phillippines Council for Agriculture and Resources Research, Los Banos, Laguna, Phillippines, pp 301-312

57. E1Faki HA, Venkataraman LV, Desikachar HSR (1984) Effect of processing on the in vitro digestibility of proteins and carbohydrates in some Indian legumes. Qual Plant Plant Fds Hum Nutr 34:127-133

58. El-Hag N, Haard NF, Morse RE (1978) Influence of sprouting as the digestibility coef- ficient, trypsin inhibitor and globulin proteins of red kidneybeans. J Fd Sci 43:t874-1875

*59. Elias LG, Hernandez M, Bressani R (1976) The nutritive value of precooked legume flours processed by different methods. Nutr Rep Intl 14:385

60. Elias LG, DeFernadez DG, Bressani R (1979) Possible effects of seed coat polyphenolics on the nutritional quality of bean protein. J Fd Sci 44:524-527

61. Ellendrieder G, Geronazzo H, DeBojarski AB (1980) Thermal inactivation of trypsin inhibitors in aqueous extracts of soybeans, peanuts and kidneybeans - - presence of sub- stance that accelerate inactivation. Cereal Chem 57:25-27

* Originals not available.

220

62. Entlicher G, Kocourek J (1975) Studies on phytohemagglutinins. XXIV. Isoelectric point and hybridization of the pea (Pisum sativum) isophytohemagglutinins. Biochim Biophys Acta 393:165-169

63. Entlicher G, Kostir JV, Kocourek J (1970) Studies on phytohemagglutinins. III. Isolation and charcterization of hemagglutinins from the pea (Pisum sativum). Biochim Biophys Acta 221:272~81

64. Etzler ME, Branstractor ML (1974) Differential localization of cell surface and secretory components in rat intestinal epithelium by use of lectin. J Cell Biol 62:329-343

65. Evans RJ, Pusztai A, Watt WB, Bauer DH (1973) Isolation and properties of protein fractions from navybeans (Phaseolus vulgaris) which inhibit growth of rats. Biochim Biophys Acta 303:175-184

66. Faber JL, Zimmerman DR (1973) Evaluation of infrared roasted and extruder-processed soybeans in baby pig diets. J Anita Sci 36:902-907

67. Fenwick DE, Oakenfull DG (1981) Saponin content of soybeans and some commercial soybean products. J Sci Fd Agr 32:273-278

68. Fenwick DE, Oakenfull DG (1983) Saponin content of food plants and some prepared foods. J Sci Fd Agr 34:186-191

69. Fliegerova O, Selvetova A, Ticha M, Kocourek J (1974). Studies on phytohemagglutinins. XIX. Subunit structure of the lentil isophytochemagglutinins. Biochim Biophys Acta, 35 I: 416-426

70. Frohlich AA, Marquardt RR (1983) Turnover and hydrolysis of vicine and convicine in avian tissues and digesta. J Sci Fd Agr 34:153-163

71. Gardiner EE, Marquardt RR, Kemp G (1982) Variation in vicine and convicine concentration of fababean genotypes. Canad J P1 Sci 62:589-592

72. Gatehouse AMR, Deway GM, Dove J, Fenton KA, Pusztai A (1984) Effect of seed lectins from Phaseolus vulgaris on the development of larvae of Callosobruchus maculatus mechanism of toxicity. J Sci Fd Agr 35:373-380

73. Gennis LS, Cantor CR (1976) Double-headed protease inhibitors from black-eyed peas. I. Purification of two new protease inhibitors and the endogenous protease by affinity chromatography. J Biol Chem 251:734-740

*74. Gillespie JM, Blagrove RJ, Kortt AA (1981) Characterization of wingedbean seed proteins. Proc 2nd Intern Syrup 'Developing the potential of the Winged Bean', 19-23 January, Colombo, Sri Lanka

75. Goldstein JL, Swain T (1965) The inhibition of enzymes by tannins. Phytochem 4:185-192 76. Goldstein IJ, Hayes CE (1978) The lectins: Carbohydrate-binding proteins of plants and

animals. Adv Carbohyd Chem Biochem 35:127-340 77. Goldstein IJ, Reichert CM, Misaki A (1974) Interaction of concanavalin A with model

substrates. Ann NY Acad Sci 234:283-296 78. Grant G, Greet F, Mckenzie NH, Pusztai A (1985) Nutritional response of mature rats to

kidneybean (Phaseolus vulgaris) lectins. J Sci Fd Agr 36:409-414 79. Gross R (1982) Composition and protein quality of wingedbean (Psophoearpus te-

tragonolobus). Qual Plant Plant Fds Hum Nutr 32:117-124 80. Gupta AK, Deodhar AD (1975) Variation in trypsin inhibitor (TI) activity in soybeans

(Glycin max.). Indian J Nutr Dieter 12:81-84 81. Gupta AK, Wahie N, Deodhar AN (1976) Protein quality and digestibility in vitro of

vegetable and grain type soybeans. Indian J Nutr Dietet 13:244-251 82. Gupta DP, Gupta AK (1977) Biochemical evaluation and cooking qualities of some soybean

varieties grown at different locations of Madhya Pradesh (India). Curr Agric 1:50-55 83. Gupta K, Wagle DS (1978) Antinutritional factors of Phaseolus mungoreous (Phaseolus

mungo vat. M~-I Phaseolus aureus var. Tl). J Fd Sci Technol (India) 15:133-136 84. Gupta K, Wagle DS (1980) Changes in antinutritional factors during germination in


221

100.

iOI.

102.

103.

104.

105.

106.

107.

"108.

109.

110.

Phaseolus mungoreous, a cross between Phaseolus mungo (M t -1) and Phaseolus aureus (T1). J Fd Sci 45:394-397

85. Gupta YP (1980) Khesari dal consumption - - a health hazard. Indian Farming 30:7-9 86. Gupta YP (1981) Toxic substances in raw pulses. Indian Farming, 31: 9-I t 87. Gupta YP (I981) Flatulence and digestibility of pulses. Pulse Crops Newsletter, 1(3): 63 88. Gupta YP (1982) Nutritive value of food legumes. In: Arora SK (ed.) Chemistry and

Biochemistry of Legumes, pp 287-327 New Delhi: Oxford & IBH Publishing Co 89. Gupta YP (1983) Neurotoxin in khesari dal (Lathyrus sativus) Intern J Trop Agri 1:175-185 90. Hafez YS, Mohamed AI (1983) Presence of nonprotein trypsin inhibitors in soy and

wingedbeans. J Fd Sci 48:75-76 91. Hafez YS, Mohamed AI (1983) Biochemical study on the nonprotein trypsin and chymo-

trypsin inhibitors in soybeans. J Fd Sci 48:1265-1268 92. Hamblin J, Kent SP (1973) Possible role of Phytohemagglutinin in Phaseolus vulgaris.

Nature (New Biol Ser), 245:28 30 93. Hartman CP, Saroja P (197t) A chemical method for detection of khesari dal in tur dal or

bengatgram in presence of metanil yellow. J Proc Instn (7hem (India), 43:71-73 94. Hartman CP, Divakar NG, Nagarajarao UN (1974) A study on Lathyrus sati~ls. Indian J

Nutr Dietet 11:178-191 95. Haynes R, Feeney RE (1967) Fractionation and properties of trypsin and chymotrypsin

inhibitors from limabeans. J Biol Chem 242:5378-5385 96. Hegazy MI, Marquardt RR (1983) Development of a simple procedure for the complete

extraction of vicine and convicine from fababeans (Vieiafaba L.). J Sci Fd Agr 34:100-108 97. Hegazy MI, Marquardt RR (1984) Metabolism of vicine and convicine in rat tissues.

Absorption and excretion patterns and sites of hydrolysis. J Sci Fd Agr 35:139-146 98. Hegazy MI, Read WWC (1974) Method for the determination of vicine in plant material

and in blood. J Agric Fd Chem 22:570-571 99. Hernandez-Infante M, Herrador-Pena G, Sotelo-Lopez A (1979) Nutritive value of two

different beans (Phaseolus vulgaris) supplemented with methionine. J Agric Fd Chem 27: 965-968 Hilderbrand DF, Hettiarachchy NS, Hymowitz T, Erdman JW (1981) Electrophoretic separation and properties of wingedbean seed trypsin inhibitor. J Sci Fd Agric 32:443-450 Honig DH, Hockridge ME, Gould RM, Rackis JJ (1983) Determination of cyanide in soybeans and soybean products. J Agric Fd Chem 31:272-275 Hobday SM, Thurman DA, Barber DJ (1973) Proteolytic and trypsin inhibitory activities in extracts of germinating Pisum sativum seeds. Phytochem 12:1041-1046 Honavar PM, Shih CV, Liener IE (1962) Inhibition of the growth of rats by purified hemagglutinin fractions isolated from Phaseolus vulgaris J Nutr 77:109 114 Hova EL, King S (1979) Trypsin inhibitor contents of lupine seeds and other grain legumes. New Zealand J Agri Res 22:41-42 Inamdar AN, Sohonie K (1961) Studies on nutritive value of doublebean. Part I. Digestibil- ity of doublebean flour in vitro. Ann Biochem 21:191-198 Jaffe WG (1980) Phytohemagglutinins In: Liener IE (ed.) Toxic constituents of Plant Foodstuffs, 2nd edn., pp 69 101. New York: Academic Press Jaffe WG, Gomez MJ (1975) Beans of high or low toxicity. Qual Plant Plant Fds Hum Nutr 24:359-365 Jaffe WC, Korte R (1976) Nutritional characteristics of the wingedbean in rats. Nutr Pep Intl 14:449-455 Jaffe WG, VegaLette CL (1968) Heat labile growth inhibitory factors in beans (Phaseolus vulgaris). J Nutr 94:203-210 Jaffe WG, Brucher O, Palozza A (I 972) Detection of tbur types of specific phytohemagglutinins in different lines of beans (Phaseolus vulgaris), Z Immunitat Forsch t42:439-447


222

111. Jaffe WG, Levy A, Gonzalez DI (1974) Isolation and partial characterization of bean phytohemagglutinins. Phytochem 13:2685 2693

112. Jammalian J, Aylward F, Hudson BJF (1976) Favism inducing toxins in broadbeans (Vicia faba). Examination of bean extracts for pyrimidine glucosides. Qual Plant Plant Fds Hum Nutr 26:331-339

113. Janzen DH, Juster HB, Liener IE (1976) Insecticidal action of the phytohemagglutinins in blackbeans on a bruchid beetle. Science, 192:795 796

114. Jaya TV, Venkatraman LV (1981) Influence of germination on the carbohydrate digestibility (in vitro) of chickpea (Cicer arietinum) and greengram (Phaseolus aureus). Indian J Nutr Dietet 18:62-68

115. Jayne-Williams D J, Hewitt D (1972) The relationship between the intestinal microflora and the effects of diets containing raw navybeans (Phaseolus vulgaris) on the growth of Japanese Quail (Corturnix coturnia japonica). J Appl Bact 35:331-344

116. Jayne-Williams D J, Burgess CD (1974) Further observation on the toxicity of navybeans (Phaseolus vulgaris) for Japanese quail (Corturnix coturnix Japonica). J. Appl Bact 37: 149-169 Jindal S, Soni GL, Singh R (1982) Effect of binding of lectins from lentils and peas on the intestinal and hepatic enzymes of albino rats. J Plant Foods, 4:95 Kaifu R, Osawa T, Jeanloz RW (1975) Carbohydr Res 40:111-117 Kakade ML, Evans RJ (1965) Nutritive value of navybeans (Phaseolus vulgaris). Brit J Nutr 19:269 276 Kakade ML, Evans RJ (1966) Effect of soaking and germinating on the nutritive value of navybeans. J Fd Sci 31:781-783 Kakade ML, Simons N, Liener IE (1969) An evaluation of natural Vs synthetic substrates for measuring the antitryptic activity of soybean samples. Cereal Chem 46:518 526 Kakade ML, Simons N, Liener IE, Lambart JW (1972) Biochemical and nutritional assessment of different varieties of soybeans. J Agric Fd Chem 20:87-90 Kakade ML, Rackis JJ, Mcghee JE, Puski G (1974) Determination of trypsin inhibitor activity of soyproducts: A collaborative analysis of an improved procedure. Cereal Chem 51: 376-382

123. Kamalakannan V, Sathyamoorthy V, Motlag DE (1984) Purification, charcterization and kinetics of a trypsin inhibitor from blackgram (Vigna mungo). J Sci Fd Agr 35:199-206

124. Kapoor AC, Gupta YP (1978) Trypsin inhibitor activity in soybean seed as influenced by stage of its development and different treatments and the distribution in its anatomical parts. Indian J Nutr Dietet 15:429-433 Katsuki Y, Yasuda K, Ueda K, Naoi Y (1978) Ann Rep Tokyo Metrop Res Lab Bublic Health, 29:261 Kaur P, Bhatia IS (1984) Proteins and trypsin inhibitor activity of guar (Cyamopsis te- tragonoloba L. Taub.) seed. J Sci Fd Agr 35:987-995 Kies C (1981) Bioavailability--a factor in protein quality. J Agric Fd Chem 29:435-440 Kies C, Fox JM (1977) Dietary hemicellulose interactions influencing serum lipid patterns and protein nutritional status of adult men. J Fd Sci 42:440-443 King TP, Pusztai A, Clark EMW (1980) Kidneybean (Phaseolus vulgaris) lectin-induced lesions in rat small intestine. I. Light microscopic studies. J Comp Path 90:585 595 Kortt AA (1979) Isolation and characterization of the trypsin inhibitors from wingedbean seed (Psophocarpus tetragonolobus (L) DC). Biochim Biophys Acta, 577:371 382 Kortt AA (1980) Isolation and properties ofa chymotrypsin inhibitor from wingedbean seed (P. tetragonolobus (L) DC). Biochim Biophys Acta, 624:237 248 Kortt AA (1981) Specificity and stability of the chymotrypsin inhibitor from wingedbean seed (P. tetragonolobus (L) DC). Biochim Biophys Acta 657:212 221 Kortt AA (1983) Comparative studies on the storage proteins and anti-nutritional factors

116(a).

"117. 118.

119.

120.

121.

122.

"125.

126.

127. 128.

"129.

130.

131.

132.

133.


133(a).

134.

135.

136.

137.

138.

139.

140.

"141.

142.

143.

144. 145.

146.

147.

"148.

149.

150.

151.

152.

153.

154.

155.

223

from seeds of Psophocarpus tetragonolobus (L) DC from south-east Asian countries. Qual Plant Plant Fds Hum Nutr 33 :2940 Kortt AA, Caldwell JB (1985) Isolation of the acidic and basic lectins from wingedbean seed (Psophocarpus tetragonolobus (L) DC). J Sci Fd Agr 36:863-870 Krogdahl A, Holm H (1979) Inhibition of human and rat pancreatic proteinases by crude and purified soybean proteinase inhibitors. J Nutr 109:551 558 Kute LS, Kadam SS, Salunkhe DK (1984) Changes in sugars, starch and trypsin inhibitor activity in wingedbean (Psophocarpus tetragonolobus (L) DC) during seed development. J Fd Sci 49:314-315 Laskowski M Jr, Ikunoshin K (1980) Protein inhibitors of proteinases. Ann Rev Biochem 49:593-626 Laxmann J, Padmanabhan G (1974) Effect of/~-N-oxalyl-L-e,/%diaminopropionic acid on glutamate uptake by synaptosomes. Nature (Lond), 249:469470 Levine D, Kaplan M J, Greenway PJ (1972) The purification and charcterization of wheat- germ agglutinin. Biochem J 129:847-856 Liener IE (1951) The intraperitoneal toxicity of concentrates of the soybean trypsin inhibitor. J Biol Chem 193:183-191 Liener IE (1953) Soyin, a toxic protein from the soybean. I Inhibition of rat growth. J Nutr 49:527-539 Liener IE (1958) Effect of heat on plant proteins. In: Altschul AM (ed.) Processed Plant Protein Foodstuffs, p. 79. New York: Academic Press Liener IE (1973) Toxic factors associated with legume proteins. Indian J Nutr Dietet 10: 303-322 Liener IE (1974) Phytohemagglutinins: Their nutritional significance. J Agric Fd Chem 22: 17-22 Liener IE (1976) Phytohemagglutinins. Ann Rev Plant Physiol 27:291-319 Liener IE (1979) Significance for humans of biologically active factors in soybeans and other food legumes. J Am Oil Chem Soc 56:121-129 Liener IE (1982) Toxic Constituents in Legumes In Arora SK (ed.) Chemistry and Bioche- mistry of Legumes, Oxford & IBH Publ. Co., New Delhi, pp. 217-257 Liener IE, Kakade ML (1980) Protease inhibitors. In: Liener IE (ed.) Toxic Constituents of Plant Foodstuffs, 2rid edn pp 7-68. New York: Academic Press Lin JY, Lee SW, Lin TI, Ling KH (1977) Divicine and favism. J Chinese Biochem Soc 6: 92-95 Lis H, Sharon N (1973) The biochemistry of plant lectins (phytohemagglutinins). Ann Rev Biochem 42:541-574 Lis H, Sharon N, Katchalski E (1964) Isolation of a mannose-containing glycopeptide from soybean hemagglutinin. Biochim Biophys Acta, 83:376378 Lotan R, Siegelman HW, Lis H, Sharon N (1974) Sub-unit structure of soybean agglutinin. J Biol Chem 249:1219 1224 Mager J, Glaser G, Razin A, Izak G, Noam M (1965) Metabolic effects of pyrimidines derived from fababean glycosides on human erythrocytes deficient in glucose-6-phosphate dehydrogenase. Biochem Biophys Res Commun 20:235-240 Mager J, Razin A, Hershko A (1980) Favism In: Liener IE (ed.) Toxic Constituents of Plant

foodstuffs, 2nd edn. pp 293-318 New York: Academic Press. Malathi K, Padmanabhan G, Sarma PS (1967) Studies on biosynthesis of/~-N-oxalyl-L-c~, /~-diaminopropionic acid, the Lathyrus sativus neurotoxin. Biochim Biophys Acta, 141: 71-78 Malathi K, Padmanabhan G, Sarma PS (1968) Oxalylation of some amino acids by an enzyme preparation from Lathyrus sativus. Indian J Biochem 5:184-185


224

156.

157.

'158.

159.

Malathi K, Padmanabhan G, Sarma PS (1970) Biosynthesis of//-N-oxalyl-L-~,/~-diaminopropionic acid, the Lathyrus sativus neurotoxin. Phytochem 9:1603-1610 Manage L, Sohonie K (1972) Proximate composition, amino acid make-up and in vitro proteolytic digestibility of horsegram. J Fd Sci Tech (India), 9:35-36 Manage L, Joshi A, Sohonie K (1972) Toxicity to rats and mice of purified phytohemagglutinins from four Indian legumes. Toxicon 10:89-91 ManciniFilho J, Lajolo FM, Vizeu DM (1979) Lectins from red kidneybeans: Radiation effect on agglutinating and mitogenic activity. J Fd Sci 44:1194-1196

160. Marik T, Entlicher G, Kocourek J (1974) Studies on phytohemagglutinins. XVI. Subunit structure of the pea isophytohemagglutinins. Biochim Biophys Acta, 336:53-61

161. Marquardt RR, Ward AT (1979) Chick performance as affected by autoclave treatment of tannin-containing and tannin-free cultivars of fababeans. Canad J Anita Sci 59:781 789

162. Marquardt RR, Frohlich AA (1981) Rapid reversed-phase high performance liquid chromatographic method for the quantitation of vicine, convicine and related compounds. J. Chrom~ttogr. 208:373-379

163. Marquardt RR, Muduuli DS, Frohlich AA (1983) Publication and some properties ofvicine and convicine isolated from fababean (Vicia faba L.) protein concentrate. J. Agric. Fd. Chem. 31:839-843

164. Miller JB, Hsu R, Heinrikson R, Yachnin S (1975) Extensive homology between the subunits of the phytohemagglutinin mitogenic proteins derived from Phaseolus vulgaris (glycoprotein/lymphocyte). Proc. Natl. Acad. Sci. (USA), 72:1388-1391

165. Mirelman D, Galun E, Sharon N, Lotan R (1975) Inhibition of fungal growth by wheat germ agglutinin. Nature (Lond.), 256:414-416

166. Mohan VS, Nagarajan V, Gopalan C (1966) Simple practical procedures for the removal of toxic factors in Lathyrus sativus (Kesari dhal). Indian J Med Res 54:410-414

167. Montogomery RD (1980) Cyanogenetic glycosides. In: Liener IE (ed.) Toxic Constituents of Plant Foodstuffs, 2nd edn., pp 143-158. New York: Academic Press

168. Muduuli DS, Marquardt RR, Guenter W (198 I) Effect of dietary vicine on the productive performance of laying chickens. Canad J Anita Sci 61:75%764

169. Muduuli DS, Marquardt RR, Guenter W (1982) Effect of dietary vicine and vitamin E supplementation on the productive performance of growing and laying chicken. Brit J Nutr 47:53-60

170. Murti VVS, Seshadri TR, Venkitasubramanian TA (1964) Neurotoxic compounds of the seeds of Lathyrus sativus. Phytochem 3:73-78

171. Nagarajan V, Gopalan C (1968) Variation in the neurotoxin (/~-N-oxalyl aminoalanine) content in Lathyrus sativus samples from Madhya Pradesh. Indian J Med Res 56:95-99

172. Nagarajan V, M ohan VS (1967) A simple and specific method for detection of adulteration with Lathyrus sativus Indian J Med Res 55:1011 1014

173. Nagarajan V, Mohan VS, Gopalan C (1965) Toxic factors in Lathyrus sativus Indian J Med Res 53:269-272

174. Nagarajan V, Mohan VS, Gopalan C (I 966) Further studies on the toxic factor in Lathyrus sativus: Potentiation of a toxic fraction from the seeds by some amino acids. Indian J Biochem 3:130-131 Natarajan KR (1976) India's poison peas. Chemistry, 49:12-13 Noor MI, Bressani R, Elias LG (1980) Changes in chemical and selected biochemical components in cotyledons of germinating Phaseolus vulgaris mungo seeds. Bot Mag (Tokyo), 92:325 Oakenfull D (1981) Saponins in food- -a review. Food Chem 7" 19-40 Oakenfull DG, Fenwick DE, Hood RL, Topping DL, Illman RJ, Storer GB (1979) Effects of saponins on bile acids and plasma lipids in the rat. Brit J Nutr 42:209-216

179. Olaboro G, Campell LD, Marquardt RR (1981) Influence of fababean fractions on egg

175. *176.

"177. 178.


225

weight among laying hens fed test diets for a short time period. Canad J Anim Sci 61: 751-755

180. Olaboro G, Marquardt RR, Campell LD (1981) Isolation of the egg weight depressing factor in fababeans (Viciafaba L. var. minor). J Sci Fd Agr 32:1'074-1080

181. Olaboro G, Marquardt RR, Campell LD, Frohlich AA (1981) Purification, identification and quantification of an egg weight depressing factor (vicine) in fababeans (Viciafaba L.) J Sci Fd Agr 32:1163-1171

"182. Olsnes I, Pihl A (1978) Abrin and ricin--two toxic lectins. Trend Biochem Sci 3:7-10 "183. Onayemi O, Pond WG, Krook L (1976) Effects of processing on the nutritive value of

cowpeas (Vigna sinensis) for the growing rat. Nutr Rep Intl 13:197 184. Padmanabhan G, Cheema PS, Malathi K, Laxmanan J (1971) Neurolathyrism. J Sci Industr

Res 30:716-720 185. Palmer R, Mclntosh A, Pusztai A (1973) The nutritional evaluation of kidneybeans (Pha-

seolus vulgaris). The effect of nutritional value of seed germination and changes in trypsin inhibitor content. J Sci Fd Agr 24:937444

186. Panlova M, Entlicher G, Ticha M, Kostier JV, Kocourek J (1971) Studies on phytohemagglutinins. VII. Effect of Mn 2+ and C a 2+ o n hemagglutination and polysaccharide precipita- tion by phytohemagglutinin of Pisum sativum Biochim Biophys Acta, 237:513-518

187. Pere M, Font J, Bourrillon R (I 974) Poids moleculaire et structure sous-unitaire de la lectine de Dolichos biflorus. Biochim Biophys Acta 365:40-46

188. Phadke K, Sohonie K (1962) Nutritive value of fieldbean (Dolichos lab&b). In vitro digestibility of raw and autoclaved beans. J Sci Industr Res 21C: 155-158

189. Phillips RD, Chhinnan MS, Mendoza LG (1983) Effect of temperature and moisture content on the kinetics of trypsin inhibitor activity, protein in vitro digestibility and nitrogen solubility of cowpea flour. J Fd Sci 48:1863--I867

'190. Potter JD, Topping DL, Oakenfull DG (1979) Soya saponins and plasma cholesterol. Lancet, I: 223

"191. Potter JD, Illman RJ, Calvert GD, Oakenfull DG, Topping DL (1980) Soya saponins, plasma lipids, lipoproteins and fecal bile acids: a double blind cross-over study. Nutr Rep Intl 22:521-528

I92. Prabhu KS, Saldanha K, Pattabiraman TN (1984) Natural plant enzyme inhibitors: A comparative study of the action of legume inhibitors on human and bovine pancreatic proteinases. J Sci Fd Agr 35:314-321

193. Price ML, Butler LG, Rogler JC, Featherston WR (1979) Overcoming the nutritionally harmful effects of tannin in sorghum grain by treatment with inexpensive chemicals. J Agric Fd Chem 27:441-445

194. Price ML, Hagerrnan AE, Butler LG (1980) Tannin content of cowpeas, chickpeas, pigeon- peas and mungbeans. J Agric Fd Chem 28:459-460

195. Pueppke S (1979) Purification and characterisation ofa lectin from seeds of the wingedbean, P. tetragonolobus (L) DC Biochim Biophys Acta, 581:63-70

196. Pusztai A (1966) The isolation of two proteins, glycoprotein I and a trypsin inhibitor from the seeds of kidneybean (Phaseolus vulgaris). Biochem J 101:379-384

197. Pusztai A, Watt WB (I974) Isolectins of Phaseolus vulgaris: A comparative study of fractionation. Biochim Biophys Acta 365:57-71

I98. Pusztai A, Grant G, Palmer R (1975) Nutritional evaluation of kidneybean (Phaseolus vulgaris): the isolation and partial characterization of toxic constituents. J Sci Fd Agric 26: 149-156

199. Pusztai A, Clarke EMW, King TP (1979) The nutritional toxicity of Phaseolus vulgaris lectins, Proc Nutr Soc 38:115-121

200. Pusztai A, Clark EMW, Grant G, King TP (1981) The toxicity ofPhaseolus vulgaris lectins. Nitrogen balance and immunological studies. J Sci Fd Agr 32:1037-1046


226

"201.

202.

203.

204.

Pusztai A, King TP, Clark EMW (1982) Recent advances in the study of nutritional toxicity of kidneybean lectin in rats. Toxicon 20:195 Rackis JJ (t966) Soybean trypsin inhibitors. Their inactivation during meal processing. Fd Technol 20:1482-1484 Rackis J J, McGhee JE (1975) Biological threshold levels of soybean trypsin inhibitors by rat bioassay. Cereal Chem 52:85-92 Rackls J J, McGhee JE. Liener IE, Kakade ML, Puski G (1974) Problems encountered in measuring trypsin inhibitor activity of soy flour: Report of a collaborative analysis. Cereal Sci Today, t9:513-516

205. Ramachand CN, Parekh LJ, Ramakrishnan CV (1981) Studies on neurotoxin from Lathy- rus sativus. Indian J Biochem Biophys 18 (suppl.): 140

206. Ramamani S, Subramanian N (1981) Growth depressing effects of isolated fractions from fieldbean (Dolichos lablab) on albino rats. Indian J Nutr Dietet 18:243-254

207. Rao SLN (i975) Metabolism of BOAA, the Lathyrus sativus neurotoxin. Proc Nutr Soc (India), I9:31-34

*208. Rao SLN, Sarma PS (1967) Neurotoxic action of ODAP. Biochem Pharmac 16:218 219 209. Rao SLN, Sarma PS, Mani KS, Raghunatharao TR, Sriramachari S (1967) Experimental

neurolathyrism in monkeys. Nature (Lond.) 214:610~617 210. Rasanen V, Weber TH, Grasbeck R (1973) Crystalline kidneybean leucoagglutinin. Eur J

Biochem 38:193-200 "211. Rea R, Thompson LU, Jenkins DJA (1985) Lectins in foods and their relation to starch

digestibility. Nutr Res 5:919 212. Reddy NR, Goll DE, Salunkhe DK (1983) Effects of germination on proteins, raffinose

oligosaccharides and antinutritional factors in the Great Northern beans. J Fd Sci 48: 1796-1800

213. Richardson MA (1977) A review. The proteinase inhibitors of plants and micro-organisms. Phytochem 16:159-169

214. Rockland LB, Radke TM (1981) Legume protein quality. Fd Technol 35(3): 7982 215. Roy DN, Nagarajan V, Gopalan C (1963) Production of neurolathyrism in chicks by the

injection of Lathyrus sativus concentrates. Curr Sci 32:116-118 216. Rukmini C (1968) Isolation and purification of a new toxic factor from Lathyrus sativus

Indian J Biochem 5:182-184 217. Rukmini C (I969) Structure of the new toxin from the Lathyrus sativus. Indian J Chem 7:

1062-1063 218. Salgarkar S, Sohonie K (1965) Hemagglutinins of fieldbean (Dolichos lablab). II. Effect of

feeding fieldbean hemagglutinin A on rat growth. Indian J Biochem 2:197-198 219. Sarma PS, Padmanabhan G (1980) Lathyrogens. In: Liener IE (ed.) Toxic Constituents of

Plant Foodstuffs, 2nd edn., pp. 267~91. New York: Academic Press 220. Sastry S, Srinivasan KS, Rajagopalan R (1969) Studies on the dehulling and screw process-

ing of soybean to obtain optimally processed soyflour. J Fd Sci Techno] (India), 5:189-191 221. Sathe SK, Salunkhe DK (1981) Studies on trypsin and chymotrypsin inhibitory activities,

hemagglutinating activity, and sugars in the Great Northern beans (Phaseolus vulgaris L.). J Fd Sci 46:626-629

222. Sathe SK, Salunkhe DK (1981) Investigations on wingedbean Psophocarpus tetragonolobus (L) DC) proteins and antinutritional factors, J Fd Sci 46:1389-1393

223. Satterlee LD, Kendrick JG, Marshall HF, Jewell DK, All RA, Heckman MM, Steinke HF, Larson P, Phillips RD, Sarwar G, Slump P (1982) In vitro assay for predicting protein efficiency ratio as measured by rat bioassay: Collaborative study. J Assoe Off Anal Chem 65:798--809

*224. Satwardhar PN, Kadam SS, Salunkhe DK (1981) Effects of germination and cooking on polyphenols and in vitro protein digestibility of horsegram and mothbean. Plant Fds Hum Nutr 1:71


227

*225. Seigler DS (1977) The naturally occurring cyanogenic glycosides. Proc Phytochem 4:83-120 226. Sela BA, Lis H, Sharon N, Sachs L (1973). Isotectins from waxbean with differential

agglutination of normal and transformed mammalian cells. Biochim Biphys Acta, 310: 273-277

227. Sequeira L (1978) Lectins and their role in host-pathogen specificity. Ann Rev Phytopathol 16:453-481

228. Sgarbieri VC, Whitaker JR (1982) Physical, chemical and nutritional properties of common bean proteins. Adv Fd Res 28:93

229. Sharon N, Lis H (1972) Lectins: Cell-agglutinating and sugar-specific proteins. Science, 177: 949-959

230. Simovie R, Summers JD, Bilanski WK (1972) Heat treatment of full-fat soybeans. Canad J Anita Sci 52:183-188

231. Singh U, Eggum BO (1984) Factors affecting the protein quality of pigeonpea (Cajanus cajan L.). Qual Plant Plant Fds Hum Nutr 34:273-283

232. Sitren HS, Ahmed EM, George DE (1985) In vivo and in vitro assessment of antinutritional factors in peanut and soy. J Fd Sci 50:418-423

233. Smith C, Megen WW, Twaalfhoven L, Hitchock C (1980) The determination of trypsin inhibitor levels in foodstuffs. J Sci Fd Agr 31:341 350

234. Sodipo OA, Arinze HU (1985) Saponin content of some Nigerian foods. J Sci Fd Agr 36: 407-408

235. Sohonie K, Ambe KS (1955) Crystalline inhibitors from Indian fieldbean and doublebean. Nature(Lond.), 175:508-509

236. Sohonie K, Bhanderkar AP (1954) Trypsin inhibitors in Indian foodstuffs: Part I. Inhibitors in vegetable. J Sci Industr Res (India), 13B: 500-503

237. Sohonie K, Bhanderkar AP (1955) Trypsin inhibitors in Indian foodstuffs. II. inhibitors in pulses. J Sci Industr Res (India), t4C: 101)-104

238. Somayajulu PLN, Barat GK, Prakash S, Misra BK, Srivastava YC (1975) Breeding of low toxin containing varieties of Lathyrus sativus. Proc Nutr Soc (India), 19:35-39

239. Soni GL, Singh TP, Singh R (1978) Comparative studies on the effects of certain treatments on the antitryptic activity of the common Indian pulses. Indian J Nutr Dietet 15:341-345

240. Sosulski F (1979) Organoleptic and nutritional effects of phenolic compounds on oilseed protein products: A review. J Am Oil Chem Soc 56:711-715

241. Stein MD, Howard IK, Sage HJ (1971) Studies on a phytohemagglutinin from lentil. IV. Direct binding studies of Lens culinaris hemagglutinin with simple saccharides. Arch Bioch- em Biophys 146:353-355

242. Struthers BJ, MacDonald JR, Dahlgren RR, Hopkins DT (1983) Effects on the monkey, pig and rat pancreas of soy products with varying level of trypsin inhibitor and comparison with the administration of cholecystokinin. J Nutr 113:86--87

*243. Subbalakshmi G, Ganeshkumar K, Venkatraman LV (1976) Effect of germination on the carbohydrates, proteins, trypsin inhibitor, amylase inhibitor and hemagglutinin in horsegram and mothbean. Nutr Rep Intl 13:31-42

244. Subrahmanyan V, Narayanarao M, Swaminathan M (1957) Lathyrism. Fd Sci 6:156--160 245. Sumanthi S, Pattabiraman TN (1976) Natural plant enzyme inhibitors of seeds. Indian J

Biochem Biophys 13:52-56 246. Sumanthi S, Pattabiraman TN (1979) Natural plant enzyme inhibitors. VI. Studies on

trypsin inhibitors of Colocasia antiquorum tubers. Biochim Biophys Acta, 566:115-127 247. Takahashi T, Yagi T, Oda T, Liener IE (1974) The dissociation of waxbean hemagglutinin

by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. Agric Biol Chem 38: 865-867

248. Tamir M, Alumot E (1969) Inhibition of digestive enzymes by condensed tannins from green and ripe carobs. J Sci Fd Agr 20:199-202


228

249.

250.

251.

252.

253.

254.

Tan NH, Wong KC (1982) Thermal stability of trypsin inhibitor activity in wingedbean (P. tetragonolobus L, DC). J. Agric Fd Chem 30:1141~1143 Tan NH, Lowe ESH, Iskander M (1982) The extractability of wingedbean (P. tetragonolobus) seed trypsin inhibitors. J Sci Fd Agr 33:1327-1330 Tan NH, Rahim ZHA, Khor HT, Wong KC (1983) Wingedbean (P. tetragonolobus) tannin level, phytate content and hemagglutinating activity. J Agric Fd Chem 31:916-917 Tan-Wilson AL, Cosgriff SE, Duggan MC, Obach RS, Wilson KA (1985) Bowman-Birk proteinase isoinhibitor complements of soybean strains. J Agric Fd Chem 33:389-393 Thompson LU, Tenebaum AV, Hui H (1986) Effects of tectins and the mixing of proteins on rate of protein digestibility. J Fd Sci 51:150-152 Tomita M, Kurokawa T, Onozaki K, Ichiki N, Osawa T, Vkita T (1972) Purification of galactose-binding phytoagglutinins and phytotoxin by affinity column chromatography using sepharose. Experientia 28:84-85

*255. Toms GC (1971) In: Harborne JB, Boutter D, Turner BL (eds) Chemotaxonomy of Legumes, pp 367--462. New York: Academic Press

256. Trowbridge IS (1973) Mitogenic properties of pea lectin and its chemical derivatives (mitogenesis/chemical modification/binding affinity). Proc Natl Acad Sci (USA), 70:3650-3654

257. Trowbridge IS (1974) Isolation and chemical characterization of a mitogenic lectin from Pisum sativum. J Biol Chem 249:6004-6012

258. Tsukamoto I, Miyoshi M, Hamaguchi Y (1983) Heat inactivation of trypsin inhibitor in kintokibean (Phaseolus vutgaris). Cereal Chem 60:I94-197

259. Tsukamoto Y, Miyoshi M, Hamaguchi Y (t983) Purification and characterisation of three trypsin inhibitors from beans, Phaseolus vulgaris Kintoki. Cereal Chem. 60:282-286

260. Turner RH, Liener IE (1975) The effect of the selective removal of hemagglutinins on the nutritive value of soybeans. J Agric Fd Chem 23:484-487

261. Varshney IP (1969) Leguminous saponins. Indian J Chem 7:446-449 262. Vesely P, Entlicher G, Kocourek J (1972) Pea phytohemagglutinin selective agglutination of

tumor cells. Experientia 28:1085-1086 263. Wagner LP, Riehm JP (1967) Purification and partial characterization of a trypsin inhibitor

isolated from the navybean. Arch Biochem Biophys 121:672-677 264. Wikon BJ (1979) Naturally occurring toxicants of food. Nutr Rev 37:305-311

*265. Wilson AB, King TP, Clark EMW, Pusztai A (1980) Kidneybean lectin induced lesions in rat small intestine. 2. Microbiological studies. J Comp Path 90:597

266. Wilson KA, Laskowski M (1973) Isolation of three isoinhibitors of trypsin from garden- bean, Phaseolus vulgaris, having either lysine or arginine at reactive site. J Biol Chem 248: 756762

"267. Wing RW, Alexander JC (1971) The heating of soybean meals by microwave radiation. Nutr Rep Intl 4:387

268. Yoshida C, Yoshikawa M (1975) Purification and characterization of proteinase inhibitors from Adzukibeans (Phaseolus angularis). J Biochem 78:935-945


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