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Food Research International 43 (2010) 432442

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Food Research International


journal homepage: www.elsevier.com/locate/foodres

Review

Bioactive proteins and peptides in pulse crops: Pea, chickpea and lentil
F. Roy a, J.I. Boye b,*, B.K. Simpson a
a b

Food Science & Agricultural Chemistry Department, McGill University, Macdonald Campus, 21,111 Lakeshore Road, Ste. Anne de Bellevue, Quebec, Canada H9X 3V9 Agriculture and Agri-Food Canada, 3600 Casavant Boul, West Saint-Hyacinthe, Quebec, Canada J2S 8E3

a r t i c l e

i n f o

a b s t r a c t
Pulse crops are cool season, annually grown legume crops, which are harvested for their seeds. They are invaluable agricultural commodities which are produced and imported by many regions of the world. Pulse seeds are a valuable source of dietary protein, carbohydrates, ber and an important source of essential vitamins and minerals. Their nutritional characteristics have been associated with a reduction in the incidence of various cancers, HDL cholesterol, type-2 diabetes and heart disease. Pulses also contain protein and non-protein antinutritional factors, which may cause deleterious effects on the host when the seeds or processed seeds are consumed raw. Conversely, recent studies have demonstrated that protein antinutritional compounds such as lectins, protease inhibitors and the non-antinutritional component, angiotensin I-converting enzyme (ACE) inhibitor may have benecial properties. Lectins have been associated with reducing certain forms of cancer, activating innate defense mechanisms and managing obesity. Protease inhibitors such as trypsin and chymotrypsin inhibitors have been demonstrated to reduce the incidence of certain cancers and demonstrate potent anti-inammatory properties. Angiotensin I-converting enzyme (ACE) inhibitor has been associated with a reduction in hypertension. 2009 Elsevier Ltd. All rights reserved.

Keywords: Pulse crop Lentil Field pea Chickpea Trypsin inhibitor Chymotrypsin inhibitor Lectin Angiotensin I-converting enzyme

Contents 1. 2. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Pea, chickpea and lentil: world production and consumption. . . . . . . . . . . . . . . . . . . . . . . . . 2.1. Dry pea . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.2. Chickpea. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.3. Lentil . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Protein content of pulses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Antinutritional compounds of pulse crops . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.1. Lectin characteristics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.2. Deleterious effects of lectins on human health . . . . . . . . . . . . . . . . . . . . . . . . . . 4.3. Nutraceutical potential of pulse lectins in human health . . . . . . . . . . . . . . . . . . . 4.4. Protease inhibitors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.5. Deleterious effects of protease inhibitors on human health . . . . . . . . . . . . . . . . . . 4.6. Methods for reducing protease inhibitory compounds in pulse seeds . . . . . . . . . . . 4.7. Beneficial properties of denatured protease inhibitors on human health . . . . . . . . . Angiotensin I-converting enzyme (ACE) inhibitory peptides . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ........ ........ . . . . . . . . . . . . . . . . . . . . . ........ ........ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ........ ........ ....... ....... . . . . . . . . . . . . . . . . . . ....... ....... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ....... ....... . . . . . . . . . . . . . . . . . ....... ....... . . . . . . . . . . . . . . . . . . ....... ....... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ....... ....... ....... ....... . . . . . . . . . . . . . . . . . . ....... ....... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ....... ....... . . . . . . . . . . . . . . . . . ....... ....... . . . . . . . . . . . . . . . . . . ....... ....... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ....... ....... . . . . . . . . . . . . . . . . . 432 433 433 433 434 434 434 435 435 436 436 437 438 438 438 440 440

3. 4.

5. 6.

1. Introduction Pulse crops belong to the family of cool season, annually grown leguminous crops (Maiti & Wesche-Ebeling, 2001). They are legume crops that are harvested for their seed only and do not include legumes which are grown for oil, such as soybean (Nwok-

* Corresponding author. E-mail address: boyej@agr.gc.ca (J.I. Boye). 0963-9969/$ - see front matter 2009 Elsevier Ltd. All rights reserved. doi:10.1016/j.foodres.2009.09.002

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olo & Smartt, 1996). Consequently, pulse crops include dry pea, chickpea, lentil and lupin, along with various types of dry bean such as kidney and lima bean. These crops are produced on many continents worldwide. North America, specically Canada, and areas within Asia and the Middle East are responsible for the majority of pulse crop production and exportation. Importation of pulses occurs most frequently in populated countries such as India and Egypt, where pulses are a staple of the diet. Pulse crops are an excellent source of protein, carbohydrates, and ber, and provide many essential vitamins and minerals. Their highly nutritional properties have been associated with many benecial health-promoting properties, such as managing high cholesterol and type-2 diabetes and in the prevention of various forms of cancer. However, pulse crops and other leguminous crops also contain many antinutritional proteins, such as lectins, protease inhibitors and the non-antinutritional compound, angiotensin I-converting enzyme (ACE) inhibitor. Various deleterious effects may occur following the ingestion of raw pulse seeds or ours, such as hemagglutination, bloating, vomiting and pancreatic enlargement, due to the activity of the antinutritional compounds inside the host. Conversely, antinutritional compounds in pulses may have many benecial properties in the treatment and/or prevention of disease when properly processed. This review will focus on the global production and exportation of dry pea, chickpea and lentil, and the deleterious and benecial health effects of bioactive proteins and peptides present in pulse crops.

Pea, similar to other pulse and grain commodities, is relatively inexpensive and highly nutritious. It is high in ber (soluble and insoluble) and protein (especially rich in the essential amino acids tryptophan and lysine), low in sodium and fat, and is an excellent source of complex carbohydrates, B vitamins, folate, and minerals such as calcium, iron, and potassium. In addition, a diet high in dry pea has been demonstrated to be effective in lowering the incidence of colon cancer, type-2 diabetes, LDL-cholesterol and heart disease (Agriculture & Agri-Food Canada, 2008). Due to the nutritional value described above, pea is considered to be an important agricultural commodity. As such, it is commonly used in soups, or processed into pea our, pea starch, or pea protein concentrates. The processed pea products can be used in baked goods, soup mixes, breakfast cereals, processed meats, health foods, pastas and purees. Commercially, dry pea is available canned or dried in either whole or split pea varieties (Agriculture & Agri-Food Canada, 2008; Slinkard, Bhatty, Drew, & Morrall, 1990). In addition, pea is one of the most commonly utilized pulse crops in animal feed for poultry, sheep, cattle and swine feed rations, and is used as a feed additive in the aquaculture industry (Schatz, 2002). Finally, dry pea varieties such as AC Trapper can be seeded as a green manure crop as an alternative to summer-fallow in organic farming practices (Lawley & Shirtliffe, 2004). 2.2. Chickpea The chickpea is a member of the cool season Fabaceae (Leguminosae) family of legumes (Nwokolo & Smartt, 1996). Similar to dry pea, chickpea is one of the earliest cultivated vegetables, as it is believed to have originated in the Middle East approximately 7450 years ago (Maiti & Wesche-Ebeling, 2001). Chickpea has since been grown in temperate and semi-arid regions of the world such as Asia, Europe, Australia and North America. In 2004, 45 countries were actively producing chickpea, and together produced a total of 8.6 million metric tonnes. India was the leading producer of chickpea accounting for approximately 66% of the worlds production. Turkey was the second largest producer, producing approximately 7% of the world supply, followed by Pakistan and Iran at approximately 6% and 4%, respectively. In contrast, Canada and the United States contribute very little to the total quantity of chickpea produced worldwide, as they account for approximately 1% and less than 1% of the world production, respectively (Smith & Jimmerson, 2005a,b). The majority of the Canadian production comes from the Prairie Provinces with Saskatchewan producing approximately 81% of Canadas chickpeas, while Alberta produces the rest. Chickpea is the third most important pulse crop commodity in the world based on total production (Yust et al., 2003). In 2003, India was both the leading producer and importer of chickpea, with approximately 259 thousand metric tonnes imported (30% of the total amount imported worldwide). Pakistan and Bangladesh were the second and third largest importers of chickpea worldwide, with approximately 123 thousand (14%) and 84 thousand metric tonnnes (10%), respectively. Similar to chickpea production, the United States and Canada only marginally contributed to the total quantity of chickpea imported worldwide. In 20032004, the United States imported approximately 17 thousand metric tonnes (1%) (Smith & Jimmerson, 2005a,b), while Canada imported only 2 thousand metric tonnes (Agriculture & Agri-Food Canada, 2006a,b). There are two main commercially available types of chickpea grown worldwide: the desi and the kabuli chickpea. Desi chickpea seed is small with a dark irregular-shaped seed coat and is grown on semi-arid land. Kabuli chickpea (Garbanzo beans) is larger than desi chickpea, has a thin light-colored seed coat and is normally grown in temperate regions of the world (Agriculture & Agri-Food Canada, 2008). A variety of desi and kabuli chickpeas have been

2. Pea, chickpea and lentil: world production and consumption 2.1. Dry pea Dry pea is among the worlds oldest crops, as records indicate it was grown in the Middle East approximately 9000 years ago. In addition, it has been harvested in Europe for several thousand years (Goodwin, 2003a,b), and has since been grown in 84 countries including Australia, Canada, China and the United States (Smith & Jimmerson, 2005a,b; McKay, Schatz, & Enders, 2003). Dry pea is adapted to brown, dark brown and black soil zones in semi-arid and non-irrigated areas of the world, and has been produced in the Canadian Prairies for over 100 years (Goodwin, 2003a,b). The ve main types of pea grown worldwide are Austrian winter pea, green pea, maple pea, marrowfat pea and yellow pea. More than 60 varieties of pea have been developed for production in Canada. Canada primarily produces green and yellow pea, with only small quantities of maple, marrowfat and Austrian pea produced (Pulse Canada, 2008a,b). In 2004, 12 million metric tonnes of dry pea were produced worldwide. Canada was the leading country in pea production producing 28% of the total yield, followed by France and Russia at 14% and 10%, respectively (Smith & Jimmerson, 2005a,b). In Canada, Saskatchewan produces 68% of Canadas dry pea crop, while Alberta and Manitoba produce approximately 22% and 10%, respectively (Goodwin, 2003a,b). In addition to being the worlds largest producer of dry pea, Canada is also the leading exporter of dry pea. In 2007, Canada exported over 2 million metric tonnes of dry pea, followed by the United States and France with 469 thousand and 350 thousand metric tonnes, respectively. Conversely, in 2006, India was the largest importer of dry pea as it imported over 1.1 million metric tonnes, followed by Spain and Belgium with 663 thousand and 334 thousand metric tonnes imported, respectively. Presently, Canada and the United States are the 8th and 11th largest dry pea importing countries, importing 77 thousand and 58 thousand metric tonnes, respectively (Agriculture & Agri-Food Canada, 2008).

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developed and the characteristics of these cultivars may vary depending on the producing region. For example, some varieties such as CDC Frontier (Kabuli variety) have been developed for improved performance in the brown and dark soil zones of the Canadian Prairies. These varieties were specically designed to express certain traits such as early maturation and the ability to resist particular diseases, as this region in Canada has a short growing season and a high incidence of the soil-borne disease Ascochyta Blight (Barker, 2008). Regardless of the variety of chickpea seeded in Canada, approximately 50% of Canadas chickpea production is of the kabuli type, while the remaining 50% is of the desi type (Goodwin, 2003a,b). There is a high demand for world production, exports and imports of chickpeas due to the crops nutritional value. Chickpea is high in protein, low in fat and sodium, cholesterol free and is an excellent source of both soluble and insoluble ber, complex carbohydrates, vitamins, folate, and minerals, especially calcium, phosphorous, iron, and magnesium (Agriculture & Agri-Food Canada, 2006a,b; Nwokolo & Smartt, 1996). Chickpea is a good source of dietary protein due to its well-balanced amino acid composition and protein bioavailability (Yust et al., 2003). It is relatively inexpensive, and has been associated with the prevention of cardiovascular disease, managing type-2 diabetes and lowering LDLcholesterol levels. Insoluble dietary ber present in chickpea has been associated with reducing the incidence of colon cancer, whereas soluble ber has been demonstrated to have a benecial effect on weight loss and weight management (Agriculture & Agri-Food Canada, 2006a,b). The nutritional benets of chickpea have led to its use in various culinary applications such as hummus. In addition, chickpea is used in stews, soups and salads, and can be processed into our. 2.3. Lentil Lentil (also known as red dahl, masur, massar, masuri tillseed and split pea) is grown annually on a variety of soil types ranging from sand to clay loan soils in semi-arid regions of the world. Lentil production originated in the Near East more than 8500 years ago and has since spread to the Mediterranean, parts of Asia and was subsequently introduced into North America by the early 1900s (Oplinger, Hardman, Kaminski, Kelling, & Doll, 1990). Specically, Canada began producing lentil in 1970 (Agriculture & Agri-Food Canada, 2006a,b). Lentil is now produced in over 48 countries. In 2002, India, Turkey and Canada were the rst, second, and third largest producers, of lentil, accounting for approximately 33%, 16%, and 12%, respectively of the worlds production (Johnson & Jimmerson, 2003). Saskatchewan produces approximately 95% of the Canadian lentil crop, with the remaining 5% produced in Manitoba and Alberta (2Agriculture & Agri-Food Canada, 2006a,b). In 2001, Canada was the largest exporter of lentil with over 490 thousand metric tonnes exported, which was approximately 70% of the total quantity of lentil produced in Canada that year. Egypt was the largest importer of lentil with over 113 thousand metric tonnes. Turkey and Sri Lanka were the second and third largest importers, with over 98 and 90 thousand metric tonnes imported, respectively in 2001 (Agriculture & Agri-Food Canada, 2006; Johnson & Jimmerson, 2003). Six types of lentil are commonly grown worldwide: Eston class, French green, Laird class, red, Richlea class and Spanish brown. These six types can be further subdivided into different varieties, which are adapted to various growing environments and conditions. Some varieties are better able to adapt to certain geographical areas than others. For example, lentil varieties adapted to the Western Canadian Prairies mature earlier due the short growing season (Pulse Canada, 2008a,b). Canada produces mainly green

lentil (Laird varieties) accounting for approximately 70% of the worlds green lentil production. However, it is estimated that 70% of the lentil produced worldwide is of the red lentil type, with 25% being of the green type and 5% of the brown and other types (Agriculture & Agri-Food Canada, 2006a,b). The nutritional value of lentil and its use in a variety of culinary applications make it an important commodity in terms of production, and trade. Lentil is high in protein especially rich in lysine and leucine, low in fat, and is an excellent source of dietary ber and complex carbohydrates. Lentil also contains vitamins and minerals such as B vitamins, calcium, phosphorous and potassium, along with oleic, linoleic and palmitic acid (Adsule, 1996; Agriculture & Agri-Food Canada, 2006a,b). The nutritional characteristics of lentil have been associated with cholesterol and lipid lowering effects in humans, along with reducing the incidence of colon cancer and type-2 diabetes (Agriculture & Agri-Food Canada, 2006a,b). Lentil is not commonly used in animal feed rations; however lentil straw is frequently used as animal feed (Saskatchewan Pulse Growers, 2000). Similar to some dry pea varieties, lentil such as Indian Head black lentil, is commonly grown as a green manure crop in organic farming practices (Lawley & Shirtliffe, 2004).

3. Protein content of pulses Pulse seeds accumulate protein throughout their development, hence mature pulses seeds are normally high in protein. Chickpea, lentil, and dry pea contain approximately 22%, 28.6%, and 23.3% protein, respectively on a dry weight basis (DW) (Pulse Canada, 2004; Sotelo & Adsule, 1996). However, these percentages may vary slightly depending on plant species, variety, maturity and growing conditions. The majority of the protein found within pulse seeds is in the form of storage proteins, which are classied as albumins, globulins, and glutelins based on their solubility properties. Globulins, soluble in salt-water solutions, represent approximately 70% of the total protein found in pulses. One of two types of globulin usually predominates in pulses based on their sedimentation coefcient, vicilin or legumin which sediments at 7S or 11S, respectively. Albumins, soluble in water, account for 1020% of the total protein in pulses. Finally, glutelins, soluble in dilute acid and base, account for 1020% of the total protein found in the seeds of pulses (Duranti, 2006; Nwokolo & Smartt, 1996). Nutritionally, pulse storage proteins are relatively low in sulfur-containing amino acids, such as methionine, cysteine and tryptophan. However, the lysine content is relatively high compared to cereal crops. For this reason, combining pulse crops and grains such as rice, in the diet provides the essential amino acids required for proper human nutrition (Duranti, 2006). There are many other types of protein found in legumes including various enzymes, protease inhibitors and lectins, which are collectively known as antinutritional compounds (ANCs). The majority of these proteins are within the water-soluble albumin class of legume proteins (Nwokolo & Smartt, 1996). In addition, they are distinct from other storage proteins with respect to functionality, as they have evolved within the seed as a protective mechanism. The interest in the biological activity of pulse crop ANCs and their potential use as a nutraceutical for promoting health and wellness and preventing or managing certain diseases has increased in recent years. 4. Antinutritional compounds of pulse crops Antinutritional compounds (ANCs) are molecules that disrupt the digestion process when raw seed or our is consumed by monogastric species rendering the seed unpalatable (Domoney,

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1999). On the other hand, ruminants are not affected by protein ANCs, as they have host-specic microorganisms capable of digesting feed rich in pulse seed or pulse our. The accumulation of these antinutritional compounds within the seeds of pulses is thought to have evolved as a protective mechanism to help the plant complete its life cycle under adverse conditions, such as predation from parasites, insects, fungi and herbivores, by causing sufcient intestinal tract discomfort within the host to deter predation (Duranti, 2006). The antinutritional compounds found in pulse crops are classied into two categories: protein ANCs and non-protein ANCs (Duranti & Gius, 1997), and range in effect from relatively inoffensive polyphenols to the relatively harmful protease inhibitors. Non-protein ANCs include alkaloids (Markievicz, Kolanowka, & Gulewics, 1988), phytic acid, phenolic compounds such as tannins (Davis, 1981) and saponins (Hudson & El-Difrawi, 1979). For an in-depth review of non-protein ANCs, the reader is referred to the paper by Champ (2002) as a review of this type of ANC is beyond the scope of this paper. Protein ANCs commonly present in pulse crops include lectins or agglutinins, trypsin inhibitors, chymotrypsin inhibitors, antifungal peptide (Ye & Ng, 2002) and ribosome-inactivating proteins (Wang & Ng, 2007). The protein ANCs from chickpea, dry pea and lentil may have mild to severe deleterious effects on human and animal health. However, recent studies have identied that certain protein ANCs may also have benecial effects on human health after adequate processing procedures. 4.1. Lectin characteristics Lectins (agglutinins) are ubiquitous carbohydrate-binding proteins which are found in a wide variety of important crop plants. Lectins are dened as all plant protein possessing at least one non-catalytic domain that binds reversibly to a specic mono- or oligosaccharide (Ryan, 1990). Several hundred plant lectins have been identied, with most classied into four groups based on their molecular structure, carbohydrate binding characteristics, and biological activity (Peumans & Van Damme, 1996). The four main groups of lectins are: legume lectins, monocot mannosebinding lectins, chitin-binding lectins and the type-2 ribosomeinactivating proteins. Plant lectins are commonly found in food consumed without processing, such as fruit and vegetables (Nachbar & Oppenheim, 1980). Pea, chickpea, lentil, and other pulse crops seeds, contain higher proportions of legume lectins than other lectin groups. The biological role of legume lectins has yet to be elucidated; however it is thought that lectins act as a defense mechanism to protect the plant from predation. These protective properties of lectins have resulted in several developments in plant biotechnology and genetic engineering in exploiting lectins in certain plant cultivars as bioactive insecticides (Sadeghi, Van Damme, Peumans, & Smagghe, 2006; Shukla, Arora, & Sharma, 2005). However, despite their benecial properties as bioactive insecticides, when pulses are consumed raw, they can resist gastrointestinal digestion causing various adverse physiological effects in the host (Kelsall et al., 2002).

4.2. Deleterious effects of lectins on human health In humans and livestock, growth suppression, diarrhea, bloating, vomiting and red blood cell agglutination, when sufcient quantities of raw seed or our are consumed have been reported (Liener, Sharon, & Goldstein, 2002). While some lectins are heatlabile, others may be resistant to mild-heat treatments. Ingestion of active lectins can potentially cause red blood cell agglutination (Etzler, 1985), which may lead to complications such as hemolysis and in extreme cases, death (Liener et al., 2002). In addition to the type of heat treatment (i.e., dry versus wet heating), the hemagglutinating activity (HA) of pulses depends on the temperature of the process and the crop variety. Bender and Reaidi (1982) have reported that the HA in pulse seeds may actually increase after mild-heat treatment. In some instances, they found that heating increased the HA by sevenfold after 10 min at 80 C. Hemagglutinins also show variable heat resistance depending on cultivar (Ayyagari, Narasinga, & Roy, 1989). Lectins in chickpea, lentil and dry pea are hemagglutinins of all human blood types (A, B, AB, O), although they exhibit different agglutination activity (Table 1). Specically, the HA in lentil and eld pea is higher than in other pulse crops. Chickpea lectins have no HA in red blood cells from humans (A, B, O types), rats, rabbits or monkeys (Ayyagari et al., 1989). However, lectins from chickpea agglutinated red blood cells from cows, although not at toxic levels (Contreras & Tagle, 1974). HA of lectins is affected by several factors, such as the molecular properties of the lectin, cell surface properties, metabolic state of the cells and the conditions of the assay such as temperature and assay preparation (Lis & Sharon, 1986). In addition, the cultivar, cultivation area, and method of harvest may also affect the concentration of lectins within a particular sample (Mekbungwan, 2007). These factors may contribute to the discrepancy in the HA of chickpea compared to pea and lentil. To the authors knowledge, there have been no reports comparing the concentration of lectins in pea, lentil and chickpea, or among different cultivars of these species. The concentration of lectins in raw lentil and pea samples was 0.11 (g/kg) and 0.22 (g/kg), respectively, according to Peumans and Van Damme (1996), however chickpea was not evaluated in this study. Furthermore, the concentration of lectins in any chickpea variety remains unclear. It is possible that a lower concentration of lectins in chickpea than either pea or lentil may be responsible for its low HA. In addition to hemagglutination, mild food poisoning cases have caused vomiting, bloating, and diarrhea in humans (Duranti & Gius, 1997). Most cases are self-limiting, however some people may require intravenous uid replacement after consuming raw or inadequately processed pulse seeds. In the livestock industry, feeding monogastric animals raw pulse seeds reduces growth and performance as a result of lectins ability to bind to intestinal mucosal cells, which affects blood glucose response (Bardocz, Grant, & Pusztai, 1996). When ingested orally, lectins enhance the shedding of brush border membranes and decrease villus length in the small intestine, which in turn reduces the surface area and interferes with normal gastric secretion for nutrient absorption (DMello, 2002). Lectins also bind glycan receptors along the intestinal tract

Table 1 The agglutination activity of lectins from chickpea, lentil and eld pea. Common name Scientic name Part tested Agglutination activity on blood type A Chickpea Lentil Field pea Cicer arietinum Lens culinaris Pisum sativum Seed Seed Seed + +++ +++ B + +++ +++ AB + +++ +++ O + +++ +++

Adapted from Schuler, March 19, 2006 ABO blood type specic lectins in edible foods. http://www.owenfoundation.com/Health_Science/Lectins_in_Foods.html.

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causing discomfort and self-limiting ailments (Peumans & Van Damme, 1996). Animals fed a soybean-rich diet can develop enlarged spleen, reduced serum insulin levels and have disruption of the normal protein, fat and carbohydrate intermediary metabolism due to the activity of lectin (Sharon & Lis, 2004). The lectin activity of pea, chickpea and lentil has not been shown to produce the same pathological effects as raw soybean and kidney bean, although mild self-limiting ailments may arise in human populations and livestock. However, there are conicting reports on their toxicity. Peumans and Van Damme (1996) reported that lectins from lentil and eld pea may be harmful to humans if consumed raw, while others have found that lectins from lentil, pea and chickpea are non-toxic (Gonzalez, Mejia, & Prisecaru, 2005). A study on the lectin activity of several leguminous crops demonstrated that chickpea and pigeon pea had lectin activity, although the activity was reported to be below toxic levels (Singh, 1988). Although there is a risk of food poisoning from consuming raw pulse seeds, it should be emphasized that under standard operating processing conditions the antinutritional effect of lectins is relatively minor. 4.3. Nutraceutical potential of pulse lectins in human health Lectins present in pulses have long been considered antinutritional components, which must be denatured by thermal processing prior to human consumption. Over the past several years, scientic data has demonstrated that lectins may play a key role in preventing certain cancers, and in the activation of certain innate defense mechanisms and can be utilized as a therapeutic agent for preventing or controlling obesity (Ewen, Bardocz, Pusztai, & Pryme, 2006; Hartmann & Meisel, 2007; Lima, Sampaio, Henriques, & BarjaFidalgo, 1999; Pusztai & Bardocz 1996; Sames et al., 2001; Wang, Ng, Ooi, & Liu, 2000). The potential use of pulse lectins as a nutraceutical for controlling obesity can be attributed to their ability to resist gastric digestion and by their ability to be absorbed into the blood stream while remaining biologically active. These properties have also resulted in increased research activity in the use of pulse lectins as a cancer preventative. Although research remains in its infancy, several animal models have been described. Mice with non-Hodgkins lymphoma fed 10 mg of mistletoe lectin per day were shown to have tumors with 75% decrease in mitotic activity and 21% reduction in the nuclear area (Ewen et al., 2006). Lentil lectins have a strong effect on reducing the onset of human hepatoma (H3B) (Wang et al., 2000). Lentil agglutinins are also known to have considerable effect on human Merkel skin carcinomas (Sames et al., 2001). Pisum sativum agglutinin (PSA) has high afnity for tumor cells, which may in turn reduce the tumorigenicity of these cells (Bures, Moytycka, Bostik, Slavik, & Jirasek, 1986). Several factors can affect the efcacy of lectins in reducing the onset of certain types of tumors. The mode of action of lectins on tumor cells depends on the type of tumor, the source of lectin and its biological activity. They are thought to bind to cell membranes or cell receptors causing cytotoxicity and apoptosis. Some theories suggest agglutinin causes reduction in cell division, increase in the number of macrophages which increases the susceptibility of tumor cells to macrophage attack, and improves the host immunocompetence (Barac, Stanojevic, & Pesic, 2005). Lectins may also penetrate the cell, resulting in cancer cell agglutination and many other antitumor properties, such as the activation of certain protein kinases (Gonzalez et al., 2005). Most research on lectins/agglutinins as a potential adjunct to cancer treatment use lectin sources other than pea, lentil and chickpea. Bean varieties, amaranth, wheat, potatoes, mistletoe, vetch, soybean and many animal lectins are commonly studied. For further information regarding the use of lectins from

sources other than pulse crops, Gonzalez et al. (2005) provide a detailed review on the uses of important plant lectins as a potential cancer treatment in their paper entitled, Lectins as bioactive plant proteins: a potential in cancer treatment. Most studies investigating anticancer effects of lectins found in pulse crops have been performed with lectins from lentil and various pea varieties. In general, pea, lentil and chickpea are not well studied sources of lectins used as cancer therapeutic agents; however, these pulse lectins may have great potential as a nutraceutical used to decrease the risk of certain cancers. Lectins from various sources have been identied as active immunomodulatory agents that can enhance the immune system, such as lymphocyte proliferation, natural killer cell activity, antibody synthesis and cytokine regulation (Hartmann & Meisel, 2007). Lectins from mistletoe induce cytokine activity by mononuclear cells which can improve the effect of chemotherapeutic drugs (Ewen et al., 2006). The use of mistletoe lectin as an adjunct during cancer treatment is currently commercially available in Europe. This has inspired research to discover lectins with immunomodulatory properties from various other sources such as soybean, fungi and fruit. Soybean agglutinin lectins induce neutrophil and lymphocyte migration in vivo and activate mononuclear cells (Benjamin, Figueiredo, Henriques, & Barja-Fidalo, 1997). Recently, lectins from mushrooms (Dalloul, Lillehoj, Lee, Lee, & Chung, 2005; Wang, Liu, Ng, Ooi, & Chang, 1996) and Jackfruit (Artocarpus Integrifolia) (Coltri, Casabona-Fortunato, Panuto-Castelo, 2004) have been demonstrated to have immunomodulatory properties. The current knowledge regarding pulse crops containing lectins with immunomodulatory properties is based on pea. Pisum sativum agglutinin (PSA) studies (Lima et al., 1999) demonstrated that PSA induces immunomodulatory effects, activating spleen lymphocytes in mice. More research is needed to characterize the effects of PSA in humans, as well as the use of lentil and chickpea agglutinins as potential immunomodulatory compounds. Obesity is often considered the root cause of other illness such as heart disease, cancer and diabetes. Pulse seed lectins may also be efcacious in treating obesity. Pusztai and coworkers (Pusztai et al., 1998) demonstrated that lectins from kidney bean could decrease fat accumulation in rats, which was attributed to a decrease in insulin levels secondary to lectin activity. They also suggest that it may be possible to use bean lectin as a dietary adjunct or primary therapeutic agent in human trials to stimulate gastrointestinal function and reduce the incidence of obesity, if a safe and effective dose range were to be established (Pusztai & Bardocz, 1996). The main source of lectin used for treating/preventing obesity appears to be from bean. To the authors knowledge there are no reports suggesting that lectins isolated from chickpea, dry pea or lentil have a benecial role in treating obesity. However, since lectins isolated from closely related pulse crops such as bean have been demonstrated to treat and/or prevent obesity there may be a potential for the use of lectins isolated from other pulses as nutraceutical components. 4.4. Protease inhibitors Similar to lectins, protease inhibitors are protein ANCs that have been isolated from a wide variety of fruit, vegetable and pulse crops including pea, chickpea and lentil. In the 1970s and 1980s, considerable attention was given to protease inhibitors in pulsebased feed, as they interfered with digestion, growth and performance in domestic livestock (Champ, 2002). The level of protease inhibitors within pulses has since been considered an important parameter in determining the quality of feed and food. Two wellcharacterized protease inhibitors found in pulse seeds are trypsin and chymotrypsin inhibitors, belonging to the Bowman-Birk inhibitor (BBI) family. Protease inhibitors from the BBI family con-

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tain two active sites which inhibit the proteolytic enzymes trypsin and chymotrypsin. Due to the double inactivation of typsin and chymotrypsin, BBIs have been termed double-headed inhibitors (Domoney, Welham, & Sidebottom, 1993). When isolated from pigeon pea, trypsin and chymotrypsin inhibitors had an average molecular mass of 10,500 and 15,000 Da, respectively (Godbole, Krishna, & Bhatia, 1994). However, the molecular mass may vary between different species and environmental growing conditions. Ragg et al. (2006), Ferrasson, Quillien, and Gueguen (1997) and Smirnoff, Khalef, Birk, and Applebaum (1976), provide detailed information on the characteristics of trypsin and chymotrypsin inhibitors from pea, chickpea and lentil for further review of this subject. All protease inhibitors present in pea, chickpea and lentil belong to the BBI family, whereas protease inhibitors from soybeans may belong to the BBI or Kunitz family of protease inhibitors (Guillamon et al., 2008). The main difference between the BBI and the Kunitz inhibitors is the polypeptide make-up. The polypeptide in Kunitz inhibitors consists of 181 residues with only two disulde bridges and one active site. The BBI polypeptide consists of 70 residues and seven disulde bridges with a double-headed inhibition mechanism (Laskowski & Kato, 1980). The differences between the BBI and the Kunitz families of protease inhibitors have a bearing on their respective inhibitory activities. As protease inhibitors from pea, chickpea and lentil are from the BBI family, their inhibitory characteristics are relatively similar. However, differences in the concentration of inhibitor and their activity exist between pulse crop species (Champ, 2002) and varieties (Alonso, Orue, & Marzo, 1998). The concentration and inhibitory activity of trypsin and chymotrypsin inhibitors in three pea varieties grown in Spain were found to vary (Table 2). Protease inhibitor concentrations also demonstrated varying degrees of susceptibility to various processing treatments. This indicates that the processing technique and conditions used to denature protease antinutritional compounds may vary depending on the seed variety being processed (Alonso et al., 1998). A similar study (Morrison, Savage, Morton, & Russell, 2007) also demonstrated variable trypsin inhibitory activity between cultivars of raw peas grown in New Zealand. This study also illustrated that the decrease in activity varied between cultivars after thermal processing. Mean values of trypsin and chymotrypsin inhibitors in chickpea have also been shown to vary. It was found that the concentration of both trypsin and chymotrypsin was higher in desi than in kabuli chickpea (Singh & Jambunathan, 1981). A review by Champ (2002) compared the trypsin (trypsin inhibitory units; TIU/mg DM) and chymotrypsin (chymotrypsin inhibitory activity; CIA/g) inhibitory activity of various pulses. The results indicated that there was no TIA activity in chickpea or CIA activity in lentil or chickpea. However, other researchers found that the trypsin and chymotrypsin inhibitory activity in lentil seed meal was 1.2 and 0.9 mg/g, respectively (Weder, Hegarty, Holzner,

& Kern-Dirndorfer, 1983). Winter pulse crop varieties may also have more protease inhibitory activity than spring varieties (Champ, 2002). The results reported are variable when comparing species due to irregularities in units used and methods of analysis. A more comprehensive research is needed to analyze the activity of protease inhibition between species and varieties to further improve processing conditions and our understanding of the benecial and detrimental effects of protease inhibitors in human and animal nutrition. To date, the best characterized and studied BBIs are found in pea cultivars (Duranti & Gius, 1997), therefore more research is needed on lentil and chickpea varieties.

4.5. Deleterious effects of protease inhibitors on human health Although concentration and activity of protease inhibitors in pulse seeds are variable, detrimental effects of their inhibitory properties have been demonstrated (Champ, 2002; Duranti, 2006; Messina, 1999). Similar to lectins, protease inhibitors interfere with digestion when pulse seeds or pulse our are consumed raw. Protease inhibitors are resistant to pepsin and the acidic pH of the human digestive tract and, therefore, interfere with digestion by inhibiting trypsin and chymotrypsin through irreversible binding to the enzymes. The mechanism by which BBI and Kunitz inhibitor act has been described by Guillamon et al. (2008) as suppressing the negative feedback regulation of pancreatic secretions through the release of the hormone cholecystokinin from the intestinal mucosa. The negative feedback regulation may stimulate pancreatic hypertrophy and decrease growth and performance of the host due to reduced ability of the digestive enzymes to properly hydrolyze dietary protein, thereby decreasing the amino acid absorption and de novo proteins synthesis. The reduction in growth and performance can also be attributed to the loss of the sulfur-rich components of trypsin and chymotrypsin due to their inhibition by protease inhibitors. In addition to pancreatic enlargement secondary to the presence of protease inhibitors in raw pulse seeds, chemically induced pancreatic tumors have also been reported in some animal species from soybean trypsin inhibitors (Messina, 1999). Unlike lectins, gastrointestinal ailments such as diarrhea, bloating and vomiting associated with protease inhibitors have not been reported, and the only harmful effects on humans occurred when seeds were consumed raw or processed inadequately. The deleterious effects of protease inhibitors on humans are usually performed in in vitro trials (Duranti, 2006), however the potential benets of denatured trypsin and chymotrypsin inhibitors are being studied in in vivo models (Kennedy, 1993). In addition to affecting the host, protease inhibitors can negatively affect the properties of foodstuffs where protease enzymes are commonly used. Gel-formation, water-holding capacity, foaming and the whipping ability of a product are all negatively affected by protease inhibitors from pulse seeds (Garcia-Cerreno, 1996).

Table 2 Trypsin and chymotrypsin activity of three pea cultivars raw and after various processing treatments. Treatment Trypsin inhibitor (IU mg DM) Renata Raw seed Dehulling Soaking Germination (24 h) Germination (48 h) Germination (72 h) Extrusion 3.80 0.24 4.02 0.08 3.74 0.09 3.57 0.11 3.32 0.08 2.76 0.11 0.19 0.02 Solara 2.80 0.09 2.82 0.06 2.52 0.06 1.18 0.02 1.18 0.02 0.70 0.03 0.16 0.06 Ballet 6.32 0.23 6.47 0.08 5.56 0.19 4.64 0.14 2.81 0.06 1.55 0.09 0.34 0.02 Chymotrypsin inhibitor (IU mg DM) Renata 2.88 0.05 2.92 0.03 2.49 0.06 2.38 0.07 2.01 0.06 1.80 0.05 1.05 0.02 Solara 2.73 0.10 2.76 0.05 2.25 0.07 2.20 0.08 2.12 0.02 1.87 0.05 0.96 0.04 Ballet 4.85 0.09 4.91 0.12 4.20 0.08 4.09 0.05 3.27 0.19 2.28 0.08 1.68 0.07

DM = dry matter. IU = international units. Adapted from: Alonso et al. (1998).

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4.6. Methods for reducing protease inhibitory compounds in pulse seeds As previously mentioned, denaturation of antinutritional protease inhibitory compounds is dependant on processing conditions, pulse species and variety. Germination, extrusion cooking, dehulling and hydrothermal processing are common commercial processes used to inactivate protease inhibitors (Table 2). Studies on chicks fed processed soybean demonstrated that extrusion cooking of soybean meal at temperatures between 104 C and 120 C for 3060 s resulted in an increase in growth rates in chicks while pancreas weights remained low. The increased growth rate and the absence of pancreatic hypertrophy indicated that a specic time and temperature range was sufcient to denature the protease inhibitor in that particular soybean variety. Combining soaking with heat treatment has also been successful in decreasing the concentration of protease inhibitors. After soaking 17 different pea varieties for 18 h and then boiling them for 20 min, trypsin inhibitor content was reduced in all of the varieties by 4292% (TIU/mg DM), with an average reduction of 78% (TIU/mg DM) (Morrison et al., 2007). Other studies demonstrate variability in protease inhibitor activity after utilizing different time and temperature combinations, depending on the species, cultivar and treatment method (Alonso et al., 1998; El-Adawy, Rahma, El-Bedawy, & Sobihah, 1999; Marquez & Alonso, 1999). Agricultural practices may also have an effect on the protease inhibitor content of pulse seeds. Direct versus two-phase harvesting of different variety of pea, chickling vetch, lentil and soybean showed that two-phase harvesting reduced the trypsin inhibitory content in pea by up to 44% and up to 25% in chickling vetch. The reduction in trypsin inhibitor concentration in lentil and soybean was too small to be considered statistically relevant (Pisulewska & Pisulewski, 2000). The reduction in protease inhibitor concentrations in two-phase harvesting may be an indication that the seed accumulates protease inhibitory compounds as it reaches maturity. In a two-phase harvesting system, the plant is cut and then left in the eld to dry before the plant seed has an opportunity to accumulate the inhibitory compounds. Plant breeding programs have attempted to decrease protease inhibitor levels in pulses through genetic manipulation. France has established a protease inhibitor threshold and does not permit the registration of pea varieties with levels of trypsin inhibitors two units higher than the two ofcial control cultivars (Morrison et al., 2007). Although this policy decreases the antinutritional content in the seeds, the crops may not perform as well, resulting in decreased yields, since protease inhibitors also protect the plant from predation. Similarly, protease inhibitors also protect the seed post-harvest by protecting against fungi and other microorganisms during storage (Garcia-Cerreno, 1996). Therefore, decreasing the protease inhibitor content of pulse crops through plant breeding programs may actually be more detrimental than growing plants with a higher protease inhibitory content and subsequently inactivating the inhibitors post-farm gate through established processing techniques. 4.7. Benecial properties of denatured protease inhibitors on human health Despite the negative effects of protease inhibitors, denatured protease inhibitors have several benecial properties on human health. In 1992 BBI proteases from legume crops achieved investigational new drug status by the FDA due to their health-promoting benets in a denatured form (Kennedy, 1995). Since then numerous reports have been published on the potential health-promoting benets of protease inhibitors, such as their potential use as an anti-inammatory agent. Most research on the health benets of

protease inhibitors has been performed with soybean. A soybean extract containing soybean BBI that suppressed carcinogenesis in several animal models was developed with chymotrypsin protease inhibitor as the active component responsible for the anticarcinogenic behaviour (Kennedy, 1993). Soybean BBI suppressed colon, esophageal, liver, lung and oral carcinogenesis, and it has also been proven efcient in suppressing radiation and chemically induced carcinogenesis in in vitro transformation assays (Moy & Bilings, 1994). Studies in mice have conrmed that BBI activity was widely distributed in various tissues (blood, kidney, liver and lungs) in mice after ingestion. Human and animal broblast and epithelial cells have also been shown to internalize protease inhibitors (Moy & Bilings, 1994). These ndings indicate that BBIs are able to resist digestion and be transported to various tissues in the host. Anti-inammatory properties of protease inhibitors in pulses have also been demonstrated. Mice fed BBI concentrate demonstrated decreased inammation of the colon when ulcerative colitis was induced in comparison to mice on a standard diet (Ware, Wan, Newberne, & Kennedy, 1999). The benecial effects of BBI or other protease inhibitors as an anti-inammatory or therapeutic cancer agent depend on the pulse species and seed variety used. Protease inhibitors are suggested as potential drugs for treating various diseases such as human immunodeciency virus (HIV), hypertension and neurodegenerative disease, along with various infectious diseases, however these are often treated with synthetic peptidomimetic inhibitors rather than with natural protease inhibitors from pulses. To the best of our knowledge there have been no reports conrming the successful use of protease inhibitors from pulse crops in the treatment of the above diseases. Currently, published information regarding the use of protease inhibitors from pulse seeds as therapeutic agents is limited to the treatment of certain cancers or as an anti-inammatory agent. Further research is needed to determine if protease inhibitors derived from pulse crops can also be used in the treatment of various other diseases. Once the protease inhibitor reaches the target tissue through the blood stream, the precise mechanism of cancer and inammatory suppression is not well understood. Moy and Bilings (1994) suggest that the protease inhibitors suppress malignant transformation by inhibiting cellular enzymes involved in the induction and/or expression of the transformed phenotype. Other researchers have hypothesized that protease inhibitors suppress carcinogenesis by altering the levels of certain types of proteolytic activities, hydrolyzing activities and the expression of certain types of oncogenes that play an important role in carcinogenesis (Kennedy, 1993). Therefore, further research is needed to characterize the precise mechanism of cancer and inammation suppression.

5. Angiotensin I-converting enzyme (ACE) inhibitory peptides Angiotensin I-converting enzyme (ACE) is widely distributed in mammalian tissues, predominantly as membrane bound ectoenzymes in vascular and endothelial cells, along with several other cell types such as absorptive epithelial, neuroepithelial and male germinal cells. ACE is a zinc metallopeptidase, which is activated by chloride and has broad in vitro substrate specicity (Li, Le, Shi, & Shrestha, 2004). ACE causes high blood pressure by converting the biologically inactive angiotensin I to the potent vasoconstrictor angiotensin II, and also inactivates the vasodilator bradykinin (Hong et al., 2008) (Fig. 1). Drugs containing ACE inhibitory peptides are commonly used for treating hypertension and regulating blood pressure, however they are costly and have been associated with numerous side effects (Erdmann, Cheung, & Schroder, 2008). Since the discovery of ACE inhibitory peptides from snake venom in 1971, there has been a renewed interest in isolating ACE

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Angiotensinogen Bradykinin

Angiotensin I ACE activity

Inactivated peptide

ACE INHIBITORS Angiotensin II

Vasoconstriction
Fig. 1. Simplied schematic demonstrating the mechanism of action of ACE.

inhibitors from natural sources. ACE inhibitory peptides have been isolated from various food sources, including dairy products, casein and whey protein, soybean, eggs, sake, porcine muscle, wheat, pea and chickpea (Erdmann et al., 2008; Meisel, Walsh, Murray, & FitzGerals., 2005, chap. 13; Pedroche et al., 2002; Vermeirssen et al., 2005). ACE inhibitory peptides have proven to be effective in the prevention and treatment of hypertension, heart failure, myocardial infarctions and diabetic nephropathy in both human and animal models (Meisel et al., 2005). The efcacy of ACE inhibitory peptides is dose dependent and, therefore, dependent on their potency, which is normally measured as an IC50 value (concentration of inhibitory peptide needed for 50% inhibition of ACE activity). For example, inhibitory peptides isolated from rice, mung bean, and pea have IC50 of 18,200, 26.5 and, 0.150.23 lM, respectively (Hong et al., 2008). However, the inhibitory activity of these peptides in vitro may not have the same potency in vivo. When ingested orally, some ACE inhibitory peptides within the food matrix may be latent and must be fully or partially digested by gastrointestinal enzymes such as trypsin, chymotrypsin and pepsin. The digestion of ACE inhibitors by these enzymes may result in either inactivation or activation of the inhibitory peptide, or the peptide may remain in the latent form. Therefore, the potency measured in vitro may be vastly different from the potency in vivo. Li et al. (2004) discovered that inhibitory peptides may not always have antihypertensive activities. Peptides with a high inhibitory activity did not always translate to a decrease in blood pressure (Li et al., 2004). In addition, the amino acid sequence and the length of the inhibitory peptide chain affect the efcacy of the peptides antihypertensive properties. Inhibitory peptides with short peptide chains, usually 25 amino acids, with C-terminal proline or hydroxyproline residues have stronger inhibitory effects, as they have been shown to bind the ACE more strongly. Proline, lysine and arginine are the preferred C-terminal substrate for ACE, contributing to a more potent ACE inhibition (Erdmann et al., 2008). In addition, short peptides are more readily absorbed by the cardiovascular system than free amino acids. Larger peptides (1051 amino acids) are readily absorbed, however their ACE inhibitor potency is greatly reduced (Erdmann et al., 2008). Many food peptides and proteins derived by enzymatic hydrolysis with antihypertensive properties in vitro and in vivo have been discovered. Many in vivo studies have been performed in humans and spontaneous hypertensive rats (SHR) by intravenous and oral administration of the bioactive peptides and/or hydrolysate. Most studies have focused on the use of peptides derived from dairy products, sh, sesame seed, eggs and mung bean, among many others (Hong et al., 2008). For example, a group of hypertensive humans consuming 95 ml of Calpis sour milk daily showed

signicant reduction in blood pressure compared to placebo (Hata, Yamamoto, Ohni, Nakajima, & Takano, 1996). Fujita and Yoshikawa (1999) isolated two different peptides from thermolysin-digest of Katsuo-bushi, a traditional Japanese dish created from dried bonito. The LKPNM (100 mg/kg) and LKP (30 mg/kg) peptides decreased blood pressure in SHR by 30 and 50 mm Hg, respectively, after intravenous administration. Minimum effective doses of LKPNM and LKP were 8 and 2.25 mg/kg. Puried peptides from sesame peptide powder have been shown to substantially decrease systolic blood pressure in SHR after a single orally administered dose of either 1 or 10 mg/kg (Nakano et al., 2006). Several studies continue to demonstrate the antihypertensive characteristics of various peptides isolated from a wide variety of foodstuffs and food ingredients (Erdmann et al., 2008; Hong et al., 2008; Li et al., 2004; Meisel et al., 2005). However, to date a comparison of the ACE inhibitory properties of peptides derived from pea, chickpea and lentil to that of dairy proteins has not been adequately investigated, and further research is warranted. Investigation of pulses as a potential source of ACE inhibitory peptides has primarily focused on soybean (Kuba, Tana, Tawata, & Yasuda, 2005; Mallikarjun, Gowda, Appu, & Prakash, 2006; Wu & Ding, 2002) and mung bean, to a lesser extent (Li, Shi, Liu, & Le, 2006). However, recent research has focused on pea and chickpea (Aluko, 2008; Humiski & Aluko, 2007; Pedroche et al., 2002; Vermeirssen et al., 2005). The treatment of legumin from chickpea with alcalse produced a hydrolysate with ACE inhibitory (IC50) activity of 0.18 mg/ml. Fractionation of the hydrolysate by reverse phase chromatography also yielded six inhibitory peptides ranging in inhibitory activity (IC50) from 0.011 to 0.021 mg/ml (Yust et al., 2003). A study by Vermeirssen et al. (2005) evaluated the antihypertensive effect of pea and whey digests and investigated the success of ACE inhibitory compounds in reaching the systemic circulatory system in vitro. After intravenous administration of 50 mg protein kg -1 body weight in SHR, ACE inhibitory compounds from pea showed a transient but strong antihypertensive effect of 44.4 mm Hg, while whey digest had no effect at an equivalent dose. Peptides derived from whey were more susceptible to digestion by gastric enzymes than pea-derived ACE inhibitory peptides. In addition, pea ACE inhibitory peptides were converted to other active peptides upon gastrointestinal digestion with similar ACE inhibitory activity rather than being denatured (Vermeirssen et al., 2005). Research on the isolation and characterization of ACE inhibitory compounds from lentil varieties is limited. Work is currently ongoing in the authors laboratory to identify ACE inhibitory peptides in lentil. Presence of ACE inhibitor peptides in chickpea and pea cultivars has provided a solid foundation for the potential use of pulse seeds or pulse seed extracts in treating and preventing hypertension. However, more research is needed regarding the isolation and characterization of ACE inhibitory compounds derived from pulse crops using both in vitro and in vivo models. In addition to possessing bioactive properties that reduce hypertension, the onset of heart failure, the risk of myocardial infarctions and diabetic nephropathy, ACE inhibitors have been demonstrated to have antioxidative properties. Limited studies have demonstrated that antihypertensive drugs containing ACE inhibitors such as enalaprilat, lisinopril and captopril have antioxidative properties. These three drugs showed oxygen free radical scavenging properties by the chemiluminescence assay of oxidation of hypoxanthine by xanthine oxidase in the presence of luminal (Mira, Silva, Queiroz, & Manso, 1993). The structure of these drugs (Fig. 2) illustrates that the compounds have functional groups that react with oxygen, resulting in the scavenging of reactive hydroxyl ions through a decarboxylation reaction of the carboxyl group. Furthermore, enalaprilat and lisinopril have aromatic rings which may contribute to the compounds antioxidative actions, whereas some of the oxygen scavenging properties

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with and without encapsulation. In conclusion, additional research is needed to further investigate the antioxidative properties of pulse seeds and their potential role in human health and disease. 6. Conclusion The abundance and availability of pulse crops worldwide, coupled with their high nutritional value, have resulted in pulse crops being one of the most widely produced and consumed agricultural commodities. Pulse crops have long been known for their nutritional and health-promoting properties, such as being an excellent source of protein, ber, carbohydrates, and for their role in decreasing the risk of certain cancers, managing obesity, lowering cholesterol and type-2 diabetes. Recently, the bioactive properties of proteins and peptides derived from pulse seeds have gained increased recognition in the areas of food science and nutrition for their potential benets in treating and/or reducing the onset of disease. Lectins and protease inhibitors which were traditionally considered as protein antinutritional compounds have shown potential in the treatment and/or prevention of various cancers, obesity and hypertension which has necessitated a reconsideration of the use of the term antinutritional. Additionally, the ACE inhibitory properties of pulse peptides could make them primary therapeutic agents or adjuncts to treatment for certain cardiovascular diseases. Pulse seeds may, therefore, be potentially excellent sources of benecial bioactive proteins and peptides, and techniques for the efcient extraction and fractionation of these proteins and peptides are needed. Further research is also needed to improve our understanding of the mechanisms involved in the absorption into the blood stream, target sites and activity in various tissues of biologically active compounds derived from dry peas, chickpeas and lentils. Another area requiring research is comparative studies of the bioactivities of various pulse cultivars of the same species and between different pulse crops, in order to determine the cultivar of a specic pulse that has the greatest concentration and activity of a desired biologically active compound. References
Adsule, R. N. (1996). In E. Nwoloko & J. Smartt (Eds.), Food and feed from legumes and oilseeds (pp. 84110). Chapman & Hall Pub.. Agriculture & Agri-Food Canada (2006). Chickpeas: Situation and outlook. Bi-weekly Bulletin, 19(13). <www.agr.gc.ca> Retrieved 20.08.08. Agriculture & Agri-Food Canada (2006). Lentils: Situation and outlook. Bi-weekly Bulletin, 19(7). <www.agr.gc.ca> Retrieved 21.08.08. Agriculture & Agri-Food Canada (2008). Dry peas: Situation and outlook. Bi-weekly Bulletin, 21(2). <www.agr.gc.ca> Retrieved 20.08.08. Alonso, R., Orue, E., & Marzo, F. (1998). Effects of extrusion and conventional processing methods on protein and antinutritional factor contents in pea seed. Journal of Food Chemistry, 63(4), 505512. Aluko, R.-E. (2008). Determination of nutritional and bioactive properties of peptides in enzymatic pea, chickpea, and mung bean protein hydrolysates. Journal of AOAC International, 91, 947956. Ayyagari, R., Narasinga, B. S., & Roy, D. N. (1989). Lectins, trypsin inhibitors, BOAA and tannins in legumes and cereals and the effect of processing. Food Chemistry, 34, 229238. Barac, M. B., Stanojevic, S. P., & Pesic, M. B. (2005). Biologically active components of soybeans and soy protein products A review. APTEFF, 36, 1266. Bardocz, S., Grant, G., & Pusztai, A. (1996). The effects of phytohaemagglutinin at dietary concentrations in the growth, body composition and plasma insulin of the rat. British Journal of Nutrition, 76, 613626. Barker, B. (2008). New pea and chickpea variety for 2008. Top Crop Manager. <www.topcropmanager.com/content/view/1354/> Retrieved 20.08.08. Bender, A. E., & Reaidi, G. B. (1982). Toxicity of kidney beans (Phaseolus vulgaris) with particular reference to lectins. Journal of Plant Foods, 4, 1522. Benjamin, C. F., Figueiredo, R. C., Henriques, M. G. M., & Barja-Fidalo, C. (1997). Inammatory and anti-inammatory effect of soybean agglutinin. Brazilian Journal of Medical and Biological Research, 30, 873881. Benzie, I. F. F., & Tomlinson, B. (1998). Antioxidant power of angiotensin-converting enzyme inhibitors in vitro. British Journal of Pharmacology, 45, 168169. Bures, L., Moytycka, K., Bostik, J., Slavik, K., & Jirasek, A. (1986). The use of protein as a carrier of methotrexate for experimental cancer chemotherapy: II. Chemotherapy of gardener lymphosarchoma with pea seed lectinmethotrexate derivative. Neoplasma, 33, 409416.

Fig. 2. Chemical structure of three common antihypertensive drugs. Source: http:// www.drugbank.ca (Wishart et al., 2008).

from captopril may result from the oxidation of its sulfur group (Mira et al., 1993). In a related study of six antihypertension drugs (captopril, enalapril, fosinopril, perindopril, quinapril and ramipril), only captopril had signicant antioxidative properties when assayed by the ferric reducing antioxidant power (FRAP) assay (Benzie & Tomlinson, 1998). The comparison of antioxidative properties found in ACE inhibitor drugs to those found in raw pulse seeds or pulse our may be difcult, as ACE inhibitor potencies of pharmaceutical compounds are generally expressed in IC50 values rather than concentration. Therefore, it is difcult to denitively determine if pharmaceutical ACE inhibitors are more potent antioxidants than those derived from pulse seeds based on current information. However, it can be hypothesized that ACE inhibitor containing drugs have a higher concentration of ACE inhibitors compared to chickpea, pea and lentil, as pharmaceutical compounds are sold in capsules in a concentrated form. Furthermore, the antioxidative properties of pharmaceuticals may have a higher potency and may be metabolized more readily, as pharmaceutical compounds are typically encapsulated to protect the active ingredient from adverse conditions such as microbial activity, protease enzymes and low stomach pH. In addition, the antioxidative properties of the ACE inhibitor peptides derived from pulses may be lost due to modications to the peptide prior to being absorbed by the host. There are other components of pulse seeds, such as genistein, daidzein and other phytoestrogens, which also have been demonstrated to have strong antioxidative properties. Therefore, some of the antioxidative effects in vivo may be predominantly the result of other phenolic components and not ACE inhibitors (Mitchell, 2001). Although a small number of studies have identied certain ACE inhibitor containing drugs as having antioxidative properties, further research is needed to characterize ACE inhibitors isolated from pulse seeds. Moreover, most of the antioxidative studies described above have been performed in vitro. Further research should be undertaken to examine antioxidative potencies in vivo,

F. Roy et al. / Food Research International 43 (2010) 432442 Champ, M. M. J. (2002). Non-nutrient bioactive substances of pulses. British Journal of Nutrition, 88(3), S307S319. Coltri, K. C., Casabona-Fortunato, A., & Panunto-Castelo, A. (2004). Immunomodulation by the lectin KM+ from Artocarpus Integrifolia, a new therapeutic approach for infections where host resistance depends on Th1 response. Trends in Glycoscience and Glycotechnology, 16, S12. Contreras, S., & Tagle, M. A. (1974). Toxic factors in Chilean legumes. Journal of Arch Latinoamer Nutrition, 24, 191199. Dalloul, R. A., Lillehoj, H. S., Lee, J. S., Lee, S. H., & Chung, K. S. (2005). Immunopotentiating effect of a Fomitella fraxinea-derived lectinon chicken immunity and resistance to coccidiosis. Poultry Science Association Inc. Davis, K. R. (1981). Effect of processing on composition of tetrahymena relative nutritive value on green and yellow peas, lentils and white pea beans. Journal of Cereal Chemistry, 58, 454460. DMello, J. P. F. (2002). Contaminants and toxins in animal feeds. Food and Agriculture Organization. <www.fao.org/agrippa> Retrieved 22.08.08. Domoney, C. (1999). Inhibitor of legume seeds. In P. R. Shewry & R. Casey (Eds.), Seed protein (pp. 635655). Amsterdam Kluwer Academic Publishers. Domoney, C., Welham, T., & Sidebottom, C. (1993). Purication and characterization of Pisum seed trypsin inhibitor. Journal of Experimental Botany, 261, 701709. Duranti, M. (2006). Grain legume proteins and nutraceutical propertie. Fitoterapia, 77, 6782. Duranti, M., & Gius, C. (1997). Legume seeds: Protein content and nutritional value. Journal of Field Crop Research, 53, 3145. El-Adawy, T. A., Rahma, E. H., El-Bedawy, A. A., & Sobihah, T. Y. (1999). Effect of soaking process on nutritional quality and protein solubility of some legume seeds. Molecular Nutrition & Food Research, 44(5), 339343. Erdmann, K., Cheung, B. W. Y., & Schroder, H. (2008). The possible roles of foodderived bioactive peptides in reducing the risk of cardiovascular disease. Journal of Nutritional Biochemistry, 19(10), 643653. Etzler, M. E. (1985). Plant lectins: Molecular and biological aspects. Annual Review of Plant Physiology, 36, 209234. Ewen, S. S. W. B., Bardocz, S., Pusztai, A., & Pryme, I. F. (2006). Suppression of growth of tumor cell lines in vitro and tumors in vivo by mistletoe lectins. Histology and Histopathology, 21(3), 285299. Ferrasson, E., Quillien, L., & Gueguen, J. (1997). Proteinase inhibitors from pea seeds: Purication and characterization. Journal of Agricultural Food Chemistry, 45, 127131. Fujita, H., & Yoshikawa, M. (1999). LKPNM: A prodrug-type ACE-inhibitory peptide derived from sh protein. Immunopharmacology, 44(12), 123127. Garcia-Cerreno, F. L. (1996). Proteinase inhibitors. Trends in Foods Science & Technology, 7, 197204. Godbole, S. A., Krishna, T. G., & Bhatia, C. R. (1994). Purication and characterization of protease inhibitors from pigeon pea (Cajanus cajan (L) Millisp) seed. Journal of the Science of Food and Agriculture, 64, 8793. Gonzalez, De., Mejia, E., & Prisecaru, V. I. (2005). Lectins as bioactive plant proteins: A potential in cancer treatment. Critical Reviews in Food Science and Nutrition, 45, 425445. Goodwin, M. (2003). Crop prole for chickpeas. <www.pulsecanada.com> Retrieved 20.08.08. Goodwin, M. (2003). Crop prole for peas, Pulse Canada. <www.pulsecanada.com> Retrieved 20.08.08. Guillamon, E., Pedrosa, M. M., Burbano, C., Cuadrado, C., de Cortes Sanchez, M., & Muzquiz, M. (2008). The trypsin inhibitors present in seed of different grain legume species and cultivar. Journal of Food Chemistry, 107, 6874. Hartmann, R., & Meisel, H. (2007). Food-derived peptides with biological activity: From research to food applications. Current Opinion in Biotechnology, 18, 163169. Hata, Y., Yamamoto, M., Ohni, M., Nakajima, Y., & Takano, Y. (1996). A placebo controlled study of the sour milk on blood pressure in hypertensive subjects. American Journal of Clinical Nutrition, 64, 767771. Hong, F., Ming, L., Yi, S., Zhanxia, L., Yongquan, W., & Chi, L. (2008). The antihypertensive effect of peptides: A novel alternative to drugs? Journal of Peptides, 29, 10621071. Hudson, B. J. F., & El-Difrawi, E. A. (1979). The Sapogenins of the seeds of four lupin species. Journal of Plant Foods, 3, 181186. Humiski, L.-. M., & Aluko, R.-. E. (2007). Physicochemical and bitterness properties of enzymatic pea protein hydrolysates. Journal of Food Science, 72, S605S611. Johnson, J. B., & Jimmerson, J. (2003). Lentils. Agricultural Marketing Policy Center Montana State University, Brieng No. 60. <http://ageconsearch.umn.edu/ bitstream/29188/1/br030061.pdf> Retrieved 21.08.08. Kelsall, A., FitzGerald, A. J., Howard, C. V., Evans, R. C., Sigh, R., Rhodes, J. M., et al. (2002). Dietary lectins can stimulate pancreatic growth in the rat. International Journal of Experimental Pathology, 83(4), 203208. Kennedy, A. R. (1993). Cancer prevention by protease inhibitors. Preventative Medicine, 22, 796811. Kennedy, A. R. (1995). The evidence for soybean products as cancer preventative agents. Journal of Nutrition, 125, 733S743S. Kuba, M., Tana, C., Tawata, S., & Yasuda, M. (2005). Production of angiotensin I-converting enzyme inhibitory peptides from soybean protein with Monascus purpureus acid proteinase. Process Biochemistry, 40(6), 21912196. Laskowski, M., Jr., & Kato, L. (1980). Protein inhibitors of proteinases. Annual Reviews, 49, 593626.

441

Lawley, Y., & Shirtliffe, S. (2004). Organic agricultural center of Canada 2004 Annual report. <www.organicagcentre.ca/DOCs/OACCAnnual%20Report2004.pdf> Retrieved 20.08.08. Li, G.-. H., Le, G.-. W., Shi, Y.-. H., & Shrestha, S. (2004). Angiotensin I-converting enzyme inhibitory peptides derived from food proteins and their physiological and pharmacological effects. Nutrition Research, 24, 469486. Li, G.-. H., Shi, Y.-. H., Liu, H., & Le, G.-. W. (2006). Antihypertensive effect of alcalase generated mung bean protein hydrolysates in spontaneously hypertensive rats. European Food Research and Technology, 222(56), 14382377. Liener, I. E., Sharon, N., & Goldstein, I. J. (2002). The lectins: Properties, functions and applications in biology and medicine. Orlando, Fl: Academic Press. Lima, J. E., Sampaio, A. L. F., Henriques, M. M. O., & BarjaFidalgo, C. (1999). Lymphocyte activation and cytokine production by Pisum sativum agglutinin (PSA) in vivo and in vitro. Immunopharmacology, 41, 147155. Lis, H., & Sharon, N. (1986). Biological properties of lectins. In I. E. Liener, N. Sharon, & I. J. Goldstein (Eds.), The Lectins: Properties and applications in biology and medicine. Orlando, Fl: Academic Press. Maiti, R., & Wesche-Ebeling, P. (2001). In R. Maiti & P. Wesche-Ebeling (Eds.), Advances in chickpea science (pp. 1). Science Publishers Inc.. Mallikarjun, G. K. G., Gowda, L. R., Appu, R. A. G., & Prakash, V. (2006). Angiotensin Iconverting enzyme inhibitory peptide derived from glycinin, the 11S globulin of soybean (Glycine max). Journal of Agricultural and Food Chemistry, 54(13), 45684573. Markievicz, M., Kolanowka, A., & Gulewics, K. (1988). The chemical composition of debittered lupine seeds and their extract. Bulletin of the Polish Academy of Science. Biological Sciences(36), 13. Marquez, M. C., & Alonso, R. (1999). Inactivation of trypsin inhibitor in chickpeas. Journal of Food Composition and Analysis, 13(3), 211217. McKay, K., Schatz, B., & Enders, G. (2003). Field pea production. North Dakota State University and US Department of Agriculture. <www.ag.ndsu.edu/pubs/ plantsci/rowcrops/a1166w.htm> Retrieved 20.08.08. Meisel, H., Walsh, D. J., Murray, B., & FitzGerals, R. J. (2005). ACE inhibitory peptides. In Y. Mine & F. Shahidi (Eds.), Nutraceutical proteins and peptides in health and disease. Taylor & Francis Publishing. Mekbungwan, A. (2007). Application of tropical legumes for pig feed. Animal Science Journal, 78, 343350. Messina, M. (1999). Legumes and soybeans: Overview of their nutritional proles and health effects. The Journal of Clinical Nutrition, 70(3), 450S493S. Mira, M. L., Silva, M. M., Queiroz, M. J., & Manso, C. F. (1993). Angiotensin converting enzyme inhibitors as oxygen free radical scavengers. Free Radical Research, 19(3), 173181. Mitchell, J. H. (2001). Phytoestrogens: Involvement in breast and prostate cancer. In R. E. C. Wildman (Ed.), Handbook of nutraceuticals and functional foods (pp. 99108). CRC Press. Morrison, S. C., Savage, G. P., Morton, J. D., & Russell, A. C. (2007). Identication and stability of trypsin inhibitor isoforms in pea (Pisum sativum L.) cultivars grown in New Zealand. Journal of Food Chemistry, 100, 17. Moy, L. Y., & Bilings, P. C. (1994). A proteolytic activity in human breast cancer cell line which is inhibited by the anticarcinogenic Bowman-Birk protease inhibitor. Cancer Letters, 85, 205210. Nachbar, M. S., & Oppenheim, J. D. (1980). Lectins in the United States diet. A survey of lectins in commonly consumed foods and a review of the literature. American Journal of Clinical Nutrition(33), 23382345. Nakano, D., Ogura, K., Miyakoshi, M., Ishii, F., Kawanishi, H., Kurumazuka, D., et al. (2006). Antihypertensive effect of angiotensin I-converting enzyme inhibitory peptides from a sesame protein hydrolasate in spontaneously hypertensive rats. Journal of Bioscience, Biotechnology and Biochemistry, 70, 11181126. Nwokolo, E., & Smartt, J. (1996). In E. Nwokolo (Ed.), Food and feed from legumes and oilseeds (pp. 45). Chapman & Hall publishing. 82 and 84. Oplinger, E. S., Hardman, L. L., Kaminski, A. R., Kelling, K. A., & Doll, J. D. (1990). Alternative eld crops manual: Lentil. <www.hort.purdue.edu/newcrop/afcm/ lentil.html> Retrieved 21.08.08. n-Calle, J., Alaiz, M., Milla n, F., & Vioque, J. (2002). Pedroche, J., Yust, M. M., Giro Utilisation of chickpea protein isolates for production of peptides with angiotensin I-converting enzyme (ACE)-inhibitory activity. Journal of the Science of Food and Agriculture, 82, 960965. Peumans, W. J., & Van Damme, E. J. M. (1996). Prevalence, biological activity and genetic manipulation of lectins in foods. Trends in Food Science & Technology, 7, 132138. Pisulewska, E., & Pisulewski, P. M. (2000). Trypsin inhibitor activity of legume seeds (peas, chickling vetch, lentils, and soya beans) as affected by the technique of harvest. Animal Feed Science and Technology, 86, 261265. Pulse Canada (2004). Canadian dry peas. <http://www.pulsecanada.com> Retrieved 21.08.08. Pulse Canada (2008). Crop prole for peas. <www.pulsecanada.com/what-arepulses/peas> Retrieved 20.08.08. Pulse Canada (2008). What are pulses: Lentil (Lens culinaris). <www.pulsecanada. com/what-are-pulses/lentils> Retrieved 21.08.08. Pusztai, A., & Bardocz, S. (1996). Biological effects of plant lectins on the gastrointestinal tract: Metabolic consequences and applications. Trends in Glycoscience and Glycotechnology, 8, 149165. Pusztai, A., Grant, G., Buchan, W. C., Bardocz, S., de Carvalho, A. F. F. U., & Ewen, S. W. B. (1998). Lipid accumulation in obese Zucker rat is reduced by inclusion of raw kidney bean (phaseolus vulgaris) in the diet. British Journal of Nutrition, 79, 213221.

442

F. Roy et al. / Food Research International 43 (2010) 432442 Smith, V., & Jimmerson, J. (2005). Chickpeas (Garbanzo beans). Agricultural Marketing Policy Center Montana State University, Brieng No. 55. <www.ampc.montana.edu/briengs/brieng55.pdf> Retrieved 20.08.08. Smith, V., & Jimmerson, J. (2005). Dry peas. Agricultural Marketing Policy Center Montana State University, Brieng No. 57. <www.ampc.montana.edu/ briengs57.pdf> Retrieved 20.08.08. Sotelo, A., & Adsule, R. N. (1996). Chickpea. In E. Nwokolo & J. Smartt (Eds.), Food and feed from legumes and oilseeds (pp. 8289). London, UK: Chapman and Hall. Vermeirssen, V., Augustijns, P., Morel, N., Van Camp, J., Opsomer, A., & Verstraete, W. (2005). In vitro intestinal transport and antihypertensive activity of ACE inhibitory pea and whey digest. International Journal of Food Science and Nutrition, 56(6), 415430. Wang, H. X., Liu, W. K., Ng, T. B., Ooi, V. E., & Chang, S. T. (1996). The immunomodulatory and antitumor activities of lectins from the mushroom Tricholoma mongolicum. Immunopharmacology, 31(23), 205211. Wang, H. X., & Ng, T. B. (2007). An antifungal peptide from red lentil seeds. Peptides, 28(3), 547552. Wang, H., Ng, T. B., Ooi, V. E., & Liu, W. K. (2000). Effects of lectins with different carbohydrate-binding specicities on hepatoma, choriocarcinoma, melanoma and osteosarcoma cell lines. International Journal of Biochemistry & Cell Biology, 32(3), 365372. Ware, J. H., Wan, X. S., Newberne, P., & Kennedy, A. R. (1999). Bowman-Birk inhibitor concentrate reduces colon inammation in mice with dextran sulfate sodium-induced ulcerated colitis. Digestive Diseases and Sciences, 44(5), 986990. Weder, J. K. P., Hegarty, M. P., Holzner, M., & Kern-Dirndorfer, M. L. (1983). Trypsin and chymotrypsin inhibitors in leguminosae. Zeitschrift fr Lebensmitteluntersuchung und Forschung A, 109, 113. Wishart, D. S., Knox, C., Guo, A. C., Cheng, D., Shrivastava, S., & Tzur, D, et al. (2008). DrugBank: A knowledgebase for drugs, drug actions and drug targets. Nucleic Acids Research 36(database issue), D901-6 [PMID: 18048412]. Wu, J., & Ding, X. (2002). Characterization of inhibition and stability of soy-proteinderived angiotensin I-converting enzyme inhibitory peptides. Food Research International, 35(4), 367375. Ye, X. Y., & Ng, T. B. (2002). Isolation of a new cyclophilin-like protein from chickpeas with mitogenic, antifungal and anti-HIV-1 reverse transcriptase activities. Journal of Life Sciences, 70(10), 11291138. Yust, M. M., Pedroche, J., Giron-Calle, J., Alaiz, M., Millan, F., & Vioque, J. (2003). Production of ace inhibitory peptides by digestion of chickpea legumin with alcalase. Journal of Food Chemistry, 81, 363369.

Ragg, E. M., Galbusera, V., Scarafoni, A., Negri, A., Tedeschi, G., Consonni, A., et al. (2006). Inhibitory properties and solution structure of a potent Bowman-Birk protease inhibitor from lentil (Lens culinaris, L) seed. FEBS Journal, 273(17), 40244039. Ryan, C. A. (1990). Protease inhibitors in plants: Genes for improving defenses against insects and pathogen. Annual Review of Phytopathology(28), 425449. Sadeghi, A., Van Damme, E. J. M., Peumans, W. J., & Smagghe, G. (2006). Deterrent activity of plant lectins on cowpea weevil Callosobruchus maculatus (F.) oviposition. Journal of Phytochemistry, 67(18), 20782084. Sames, K., Shumacher, U., Halata, Z., Van Damme, E. J., Peumans, W. J., Asmus, B., et al. (2001). Lectins as bioactive plant proteins: A potential in cancer treatment. Critical Reviews in Food Science and Nutrition, 45, 425445. Saskatchewan Pulse Growers (2000). Pulse production manual 2000. <www.saskpulse.com/media/pdfs/ppm-lentil.pdf> Retrieved 21.08.08. Schatz, B. (2002). Feeding eld peas to livestock: Introduction to eld peas. Carrington Research Extension Center, North Dakota State University. EB-76 May 2002. <www.ag.ndsu.edu/pubs/ansci/livestoc/eb76w.htm> Retrieved 20.08.08. Schuler, F. M. (2006). ABO blood types specic to lectins in edible foods, Owen Foundation. <www.owenfoundation.com/Health_Science/Lectins_in_Foods. html> Retrieved 21.08.08. Sharon, H., & Lis, H. (2004). History of lectins: From hemagglutinins to biological recognition molecules. Journal of Glycobiology, 14(11), 53R63R. Shukla, S., Arora, R., & Sharma, H. C. (2005). Biological activity of soybean trypsin inhibitor and plant lectins against cotton bollworm/legume pod borer, Helicoverpa armigera. Plant Biotechnology, 22(1), 16. Singh, U. (1988). Antinutritional factors of chickpeas and pigeonpea and their removal by processing. Plant Foods for Human Nutrition, 38, 251261. Singh, U., & Jambunathan, R. (1981). Studies on desi and kabuli chickpeas (Cicer arietinum L.) cultivars: Levels of protease inhibitors, levels of polyphenolic compounds and in vitro protein digestibility. Journal of Food Science, 46(5), 13641367. Slinkard, A. E., Bhatty, R. S., Drew, B. N., & Morrall, R. A. A. (1990). Dry peas and lentil as new crops in Saskatchewan: A case study. In J. Janick, & J. E. Simon (Eds.), Advances in new crops (pp. 159163). Portland, Or: Timber Press <www.hort.purdue.edu/newcrop/proceedings1990/V1-164.html> Retrieved 20.08.08. Smirnoff, P., Khalef, S., Birk, Y., & Applebaum, S. W. (1976). A trypsin and chymotrypsin inhibitor from chick peas (Cicer arietinum). Biochemistry Journal, 157(3), 745751.

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