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Naturally occurring protease inhibitors (PI) of the Bowman-Birk type constitute a major PI family in cereal and legume seeds. The family name is derived from the names of the two investigators who characterised the first inhibitor of this... more
Naturally occurring protease inhibitors (PI) of the Bowman-Birk type constitute a major PI family in cereal and legume seeds. The family name is derived from the names of the two investigators who characterised the first inhibitor of this type, the Bowman-Birk inhibitor from soybean (BBI). These proteins have the capacity to inhibit one or more of a range of serine proteases, including the digestive enzymes trypsin and chymotrypsin. PI from this family interact with the active sites of serine proteases in a 'canonical', i.e. substrate-like, manner via exposed reactive site loops of conserved conformation within the inhibitor. Multiple BBI variants can be found within and among species. A limited number of amino acids located within the inhibitory domain is responsible for the primary functional and biological activities of BBI-like proteins. However, sequence variation in binding loops, post-translational modifications at the amino- and carboxy-terminal ends, as well as differences in the multimeric nature of the inhibitors may act in combination to influence the functional properties and the physiological role of BBI-like proteins. Recently, BBI and proteins homologous to BBI (BBI-like proteins) have emerged as highly promising cancer chemopreventive agents. BBI has been shown to be capable of preventing or suppressing carcinogenic processes in a wide variety of in vitro and in vivo animal model systems. The potential exploitation of BBI-like proteins in human health-promotion programmes will depend on elucidating in detail the molecular basis for the variation in biological activities among the many variant forms. New knowledge, derived both from the use of synthetic cyclic peptides that mimic the inhibitory loops of BBI-like proteins, and from genomic data pertaining to the structure of BBI gene classes, together facilitate the manipulation, screening and selection of appropriate variants through biotechnology.
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Glycation pattern of Bowman–Birk inhibitors from soybean affects their functional properties.
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Research Interests: Chemistry, Biology, Medicine, Food Allergy, Article, and 5 moreImmunoglobulin E, Immune system, Egg White, epitope, and Ovalbumin
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Resumen del trabajo presentado al IV Workshop Probioticos, Prebioticos y Salud: Evidencia Cientifica celebrado en Madrid (Espana) del 31 de enero al 1 de febrero de 2013.
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International audienceTrypsin / chymotrypsin inhibitors in the seeds of many legume crop species are regarded as antinutritional proteins often leading to a requirement for heat-treatment of seed products prior to their use in feed. A... more
International audienceTrypsin / chymotrypsin inhibitors in the seeds of many legume crop species are regarded as antinutritional proteins often leading to a requirement for heat-treatment of seed products prior to their use in feed. A TILLING resource developed in Pisum sativum L. (pea) was exploited to identify mutants in the major seed-expressed trypsin / chymotrypsin inhibitor gene, TI1, where the inhibition of either or both of the target enzymes may be reduced. Three lines with missense mutations in TI1, predicted to affect activity through alteration of (a) a conserved cysteine residue, (b) the P1′ serine within the active site of the chymotrypsin inhibitory domain or (c) overall charge of the carboxy-terminal domain involved in protein dimerization, were identified. The mutants were back-crossed to the parent cultivar, Cameor, to generate mutant and wild type segregant lines, where the effects of the mutations on overall activity, isoform charge and the association properties of the encoded proteins could be examined and compared. The data show that one mutation (C77Y) leads to a significant reduction in both trypsin and chymotrypsin inhibitory activity, associated with the loss of activity of two TI1 isoforms. The second and third classes of TI1 mutations showed lesser effects on overall inhibitory activity, despite their likely impact on protein structure. The resource provides potential for providing novel germplasm with improved properties for feed and food uses
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Protease inhibitors (PI) have long been considered to be antinutritional compounds in legume crops due to their ability to decrease the protein digestibility and adsorption of dietary proteins. In recent years, renewed interest in PI has... more
Protease inhibitors (PI) have long been considered to be antinutritional compounds in legume crops due to their ability to decrease the protein digestibility and adsorption of dietary proteins. In recent years, renewed interest in PI has followed the recognition that certain plant PI are effective at preventing or suppressing carcinogenic processes in a wide variety of in vitro and in vivo animal model systems (Kennedy, 1998). These reports suggest a positive contribution of PI to the nutritional value of dietary proteins from vegetable sources. The selection of plant PI for plant breeding programs aimed at human health depends on elucidation of the molecular basis for variation in their biological activity. Distinct PI have been identified in legumes with many seed inhibitors inhibiting trypsin and chymotrypsin at distinct sites (Figure 1). The significance of the two active sites is likely to be different in terms of their exploitation. The trypsin inhibitory site has been implicated in the negative effect on bioavailability of dietary proteins in food and feed (Liener, 1994) and in the protection of plants against insects (Volpicella et al., 2000) and fungi (Giudici et al., 2000). The chymotrypsin inhibitory site has been implicated in the anticarcinogenic effect of the soy-derived Bowman-Birk inhibitor (BBI) (Kennedy et. al., 1993). We are investigating the properties of a range of PI homologous to BBI that include sequence variants as well as variants that arise from post-translational processing. We have expressed two pea variant PI and their corresponding C-terminally processed forms as recombinant proteins and are assessing the antiproliferative properties of the variants on colon cancer cells using an in vitro cell assay system designed for high-throughput and rapid screening.
Research Interests: Biochemistry, Biology, Protease, Trypsin, Legume, and 3 moreProteases, Chymotrypsin, and Springer Ebooks
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This eBook is a collection of articles from a Frontiers Research Topic. Frontiers Research Topics are very popular trademarks of the Frontiers Journals Series: they are collections of at least ten articles, all centered on a particular... more
This eBook is a collection of articles from a Frontiers Research Topic. Frontiers Research Topics are very popular trademarks of the Frontiers Journals Series: they are collections of at least ten articles, all centered on a particular subject. With their unique mix of varied contributions from Original Research to Review Articles, Frontiers Research Topics unify the most influential researchers, the latest key findings and historical advances in a hot research area! Find out more on how to host your own Frontiers Research Topic or contribute to one as an author by contacting the Frontiers Editorial Office: frontiersin.org/about/contac
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Resumen del trabajo presentado al IV Workshop Probioticos, Prebioticos y Salud: Evidencia Cientifica celebrado en Madrid (Espana) del 31 de enero al 1 de febrero de 2013.
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Resumen del trabajo presentado a la 4th International Conference of Food Digestion, celebrada en Napoles (Italia) del 17 al 19 de marzo de 2015.
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Resumen del trabajo presentado al 20th International Congress of Nutrition, celebrado en Granada (Espana) del 15 al 20 de septiembre de 2013.
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Resumen del trabajo presentado al III Workshop Probioticos, Prebioticos y Salud. Evidencia Cientifica, celebrado en Castelldefels (Barcelona) del 15 al 16 de diciembre de 2011.
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Caco-2 cell monolayers to study the transport of lupin conglutin gamma across human intestinal epithelium / A. SCARAFONI, J. CAPRARO, C. MAGNI, A. CLEMENTE, M. DURANTI. - In: Proceedings of the 53rd National Meeting of the Italian Society... more
Caco-2 cell monolayers to study the transport of lupin conglutin gamma across human intestinal epithelium / A. SCARAFONI, J. CAPRARO, C. MAGNI, A. CLEMENTE, M. DURANTI. - In: Proceedings of the 53rd National Meeting of the Italian Society of Biochemistry and Molecular Biology ( ...
Research Interests: Chemistry and Epithelium
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Research Interests: Nutrition and Dietetics, Cancer, Biology, Cytotoxicity, Medicine, and 15 moreColorectal cancer, Humans, Sequence alignment, Aptitude, Soybean, Food Sciences, Alkylation, Protein isoforms, Cell Proliferation, Glycine max, Serine Protease, Cell Survival, Adenocarcinoma, Chymotrypsin, and Colorectal Neoplasms
Bowman–Birk inhibitor (BBI) from soyabeans is a naturally occurring protease inhibitor with potential anti-inflammatory and chemopreventive properties within the gastrointestinal tract (GIT). In a previous paper, we reported that... more
Bowman–Birk inhibitor (BBI) from soyabeans is a naturally occurring protease inhibitor with potential anti-inflammatory and chemopreventive properties within the gastrointestinal tract (GIT). In a previous paper, we reported that significant amounts of BBI-related proteins reach the terminal ileum functionally and biologically active. We have now investigated: (a) if soyabean BBI is biotransformed by faecal microbiota which would reduce its potential colorectal chemopreventive properties and (b) the potential influence of this protease inhibitor on the modulation of faecal microbiota. In vitro incubation studies of native soyabean BBI at a physiological level (93 μm) with mixed faecal samples of pigs for 24 h at 37°C demonstrated that BBI remains active and its intrinsic trypsin and chymotrypsin inhibitory activities were not significantly influenced by the enzymic or metabolic activity of faecal microbiota. Soyabean BBI did not affect the growth of the different bacterial groups st...
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Bowman–Birk inhibitors (BBI) from legumes, such as soyabean, pea, lentil and chickpea, are naturally occurring plant protease inhibitors which have potential health-promoting properties within the mammalian gastrointestinal tract. BBI can... more
Bowman–Birk inhibitors (BBI) from legumes, such as soyabean, pea, lentil and chickpea, are naturally occurring plant protease inhibitors which have potential health-promoting properties within the mammalian gastrointestinal tract. BBI can survive both acidic conditions and the action of proteolytic enzymes within the stomach and small intestine, permitting significant amounts to reach the large intestine in active form to exert their reported anti-carcinogenic and anti-inflammatory properties. In a previous study, we reported the ability of a recombinant form of TI1B (rTI1B), representing a major BBI isoinhibitor from pea, to influence negatively the growth of human colorectal adenocarcinoma HT29 cells in vitro. In the present study, we investigate if this effect is related directly to the intrinsic ability of BBI to inhibit serine proteases. rTI1B and a novel engineered mutant, having amino acid substitutions at the P1 positions in the two inhibitory domains, were expressed in the ...
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ABSTRACT In order to exert an effect, either local or systemic, putative beneficial dietary substances have to survive at least to some extent the digestive process within the gastrointestinal tract. For that purpose, five castrated male... more
ABSTRACT In order to exert an effect, either local or systemic, putative beneficial dietary substances have to survive at least to some extent the digestive process within the gastrointestinal tract. For that purpose, five castrated male pigs (100 ± 2 kg mean live b.w.) fitted with T-shaped ileal cannulas were used to determine the recovery up to the terminal ileum of various non-nutritional factors (NNF) from legumes (defatted soybean, raw lupin, raw chickpea and raw or autoclaved kidney beans). Kidney bean lectin (PHA) recovery within the small intestine ranged between 2.5 and 4.8 mg from 5.96 mg ingested. Two different methods were used to determine ileal digestibility of protease inhibitors (PI) in pigs. According to the enzyme-linked immunosorbent assay (ELISA) method, apparent digestibility of Kunitz trypsin inhibitor and Bowman–Birk inhibitor for pigs fed the defatted soybean-based diet were 0 and 58%, respectively. On the other hand, the apparent digestibility of PI along the small intestine, expressed in terms of trypsin inhibitory activity, was 96, 98 and 95% of activity in the diet, while data expressed in terms of chymotrypsin inhibitory activity were 95, 99 and 79% of activity in the diet for defatted soybean, raw chickpea and autoclaved kidney bean diets respectively. Differences found in ileal digestibility values of PI using both methodologies, ELISA and enzymatic inhibition, might be explained by the detection of inactive forms by specific antibodies. Intestinal apparent digestibility of phytate was 0% for defatted soybean and autoclaved kidney bean diets, whereas values for raw lupin and chickpea diets were 4.1 and 24.5%, respectively. In conclusion, significant amounts of several NNF survived the passage through the stomach and small intestine of cannulated pigs, which might have biological relevance. Copyright © 2006 Society of Chemical Industry
Research Interests: Engineering and Recovery
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Research Interests: Chemistry, Food Chemistry, Medicine, Multidisciplinary, Iu, and 4 morePlant extracts, Soybeans, CRC, and Soy milk
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Abstract: 2S albumin storage proteins are becoming of increasing interest in nutritional and clinical studies as they have been reported as major food allergens in seeds of many mono- and di-cotyledonous plants. This review describes the... more
Abstract: 2S albumin storage proteins are becoming of increasing interest in nutritional and clinical studies as they have been reported as major food allergens in seeds of many mono- and di-cotyledonous plants. This review describes the main biochemical, structural and functional properties of these proteins thought to play a role in determining their potential allergenicity. 2S albumins are considered to sensitize directly via the gastrointestinal tract (GIT). The high stability of their intrinsic protein structure, dominated by a well-conserved skeleton of cysteine residues, to the harsh conditions present in the GIT suggests that these proteins are able to cross the gut mucosal barrier to sensitize the mucosal immune system and/or elicit an allergic response. The flexible and solvent-exposed hypervariable region of these proteins is immunodominant and has the ability to bind IgE from allergic patients ´ sera. Several linear IgE-binding epitopes of 2S albumins spanning this regio...
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Resumen del trabajo presentado al 5th International Food Legumes Research Conference (IFLRC V) y al 7th European Conference on Grain Legumes celebrados en Antalya (Turquia) del 26 al 30 de abril de 2010.
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Proceedings of the I Congress PIIISA celebrado en la Estacion Experimental del Zaidin (Granada), en mayo de 2013.
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Resumen del poster presentado al 35th International Congress of the Society for Microbial Ecology and Disease (SOMED) celebrado en VAlencia (Espana) del 15 al 17 de mayo de 2012.
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The Bowman-Birk trypsin-chymotrypsin inhibitor (BBI) from soybean has been described as a potential cancer chemopreventive agent. We have compared the effects of BBI with those of two variant recombinant pea (Pisum sativum L.) seed... more
The Bowman-Birk trypsin-chymotrypsin inhibitor (BBI) from soybean has been described as a potential cancer chemopreventive agent. We have compared the effects of BBI with those of two variant recombinant pea (Pisum sativum L.) seed protease inhibitors, rTI1B and rTI2B, homologous to BBI but differing in inhibitory activity, on the growth of human colorectal adenocarcinoma HT29 cells in vitro. A significant and dose-dependent decrease in the growth of HT29 cells was observed using all protease inhibitors, with rTI1B showing the largest decrease (IC50 = 46 microM). Inclusion of the pan-caspase inhibitor, Boc-D-FMK, did not negate the effects of rTI1B or rTI2B in the cell assays. The relative effectiveness of rTI1B and rTI2B may correlate with a variant amino acid sequence within their respective chymotrypsin inhibitory domain, in agreement with a chymotrypsin-like protease as a potential target.
Research Interests: Engineering, Biology, Medicine, Cell Division, Humans, and 13 moreAgricultural, CHEMICAL SCIENCES, Seeds, Protease Inhibitors, Agricultural and Food Chemistry, Amino Acid Sequence, Recombinant Proteins, PLANT PROTEINS, PISUM SATIVUM, Adenocarcinoma, Colorectal Neoplasms, Molecular Sequence Data, and Peas
ABSTRACT: The neutral lipids composition of defatted chickpea flour and two types of protein isolates has been studied. The main compounds in neutral lipids are triacylglycerols, free fatty acids, and diacylglycerols. Other compounds... more
ABSTRACT: The neutral lipids composition of defatted chickpea flour and two types of protein isolates has been studied. The main compounds in neutral lipids are triacylglycerols, free fatty acids, and diacylglycerols. Other compounds present are wax esters, free fatty alcohols, and free sterols. The main fatty acids in neutral lipids are C 18:2 and C 18:1 among the unsaturated, and C 16:0 and C 18:0 among the saturated acids. Free and esterified alcohols range from C 16:0 to C 28:0 , the majority being those with an even number of carbon atoms. Sterols observed are β-sitosterol, campesterol, stigmasterol, and δ-5-avenasterol. Triacylglycerols are partially hydrolyzed, and the amounts of unsaturated sterols and unsaturated fatty acids are reduced as a result of the chemical treatment during production of the protein isolates. JAOCS 75, 851-855 (1998).
This study investigates the preventive effects of two pea (Pisum sativum) seed albumin extracts, either in the presence (pea seed extract) (PSE) or absence (albumin fraction from pea seed extract) (AF-PSE) of soluble polysaccharides, in... more
This study investigates the preventive effects of two pea (Pisum sativum) seed albumin extracts, either in the presence (pea seed extract) (PSE) or absence (albumin fraction from pea seed extract) (AF-PSE) of soluble polysaccharides, in the dextran sodium sulfate (DSS)-induced colitis in mice. Male C57BL/6J mice were assigned to five groups: one non-colitic and four colitic. Colitis was induced by incorporating DSS (3.5%) in the drinking water for four days, after which DSS was removed. Treated groups received orally PSE (15 g/kg•day), or AF-PSE (1.5 g/kg•day), or pure soy Bowman-Birk inhibitor (BBI) (50 mg/kg•day), starting two weeks before colitis induction, and maintained for nine days after. All treated groups showed intestinal anti-inflammatory effect, evidenced by reduced microscopic histological damage in comparison with untreated colitic mice. The treatments ameliorated the colonic mRNA expression of different pro-inflammatory markers: cytokines, inducible enzymes, metalloproteinases, adhesion molecules, and toll-like receptors, as well as proteins involved in maintaining the epithelial barrier function. Furthermore, the administration of PSE, AF-PSE or soy BBI restored bacterial counts, partially or totally, to values in healthy mice. PSE and AF-PSE ameliorated DSS-induced damage to mice, being their effects due, at least partially, to the presence of active BBI. This article is protected by copyright. All rights reserved.
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The interest for naturally-occurring oligosaccharides from plant origin having prebiotic properties is growing, with special focus being paid to supplemented products for infants. Currently, non-fructosylated α-galactooligosaccharides... more
The interest for naturally-occurring oligosaccharides from plant origin having prebiotic properties is growing, with special focus being paid to supplemented products for infants. Currently, non-fructosylated α-galactooligosaccharides (α-GOS) from peas have peaked interest as a result of their prebiotic activity in adults and their mitigated side-effects on gas production from colonic bacterial fermentation. In this study, commercially available non-fructosylated α-GOS from peas and β-galactooligosaccharides (β-GOS) derived from lactose were fermented using fecal slurries from children aged 11 to 24 months old during 6 and 24 h. The modulatory effect of both GOS on different bacterial groups and bifidobacteria species was assessed; non-fructosylated α-GOS consumption was monitored throughout the fermentation process and the amounts of lactic acid and short-chain fatty acids (SCFA) generated were analyzed. Non-fructosylated α-GOS, composed mainly of manninotriose and verbascotetraose...
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Legumes are major ingredients in the Mediterranean diet, playing an essential role in developing countries. Grain legumes, such as lentil, chickpea, pea, lupin and beans, among others, are recognized as good sources of proteins, starch,... more
Legumes are major ingredients in the Mediterranean diet, playing an essential role in developing countries. Grain legumes, such as lentil, chickpea, pea, lupin and beans, among others, are recognized as good sources of proteins, starch, fiber, vitamins and minerals for human nutrition, being an essential food crop for people worldwide. Due to their nutritional and techno-functional properties, legumes are widely used by the food industry as ingredients in a wide range of products for general and specific groups of the population, including vegetarians, diabetics or celiac patients. The Special Issue “Legumes as Food Ingredients: Characterization, Processing, and Applications” covers key aspects regarding the nutritional quality of legume flours and their derived products, as well as the health benefits of some of their bioactive components. The amounts of antinutritional components, such as certain allergens that might pose risks to sensitized consumers, are reported to be reduced b...
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Legume consumption has been reported to induce beneficial effects on obesity-associated metabolic disorders, but the underlying mechanisms have not been fully clarified. In the current work, pea (Pisum sativum L.) seed meal proteins... more
Legume consumption has been reported to induce beneficial effects on obesity-associated metabolic disorders, but the underlying mechanisms have not been fully clarified. In the current work, pea (Pisum sativum L.) seed meal proteins (albumins, legumins and vicilins) were isolated, submitted to a simulated gastrointestinal digestion, and the effects of their hydrolysates (pea albumins hydrolysates (PAH), pea legumins hydrolysates (PLH) and pea vicilin hydrolysates (PVH), respectively) on 3T3-L1 murine pre-adipocytes were investigated. The pea vicilin hydrolysate (PVH), but not native pea vicilins, increased lipid accumulation during adipocyte differentiation. PVH also increased the mRNA expression levels of the adipocyte fatty acid-binding protein (aP2) and decreased that of pre-adipocyte factor-1 (Pref-1) (a pre-adipocyte marker gene), suggesting that PVH promotes adipocyte differentiation. Moreover, PVH induced adiponectin and insulin-responsive glucose transporter 4 (GLUT4) and st...