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Takács K, Szabó EE, Nagy A, Cserhalmi Z, Falusi J, Gelencsér É. The effect of radiofrequency heat treatment on trypsin inhibitor activity and in vitro digestibility of soybean varieties (Glycine max. (L.) Merr.). JOURNAL OF FOOD SCIENCE AND TECHNOLOGY 2022; 59:4436-4445. [PMID: 36193476 PMCID: PMC9525500 DOI: 10.1007/s13197-022-05523-z] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Revised: 05/21/2022] [Accepted: 05/30/2022] [Indexed: 11/30/2022]
Abstract
Kunitz (KTI) and Bowman-Birk (BBI) trypsin inhibitors were characterized in soybean seeds. Cultivars having KTI/BBI (Pannónia Kincse, PK) or lacking KTI (Aries; Hilario; Bahia) were assessed with well-characterized soybean varieties having Ti-a or ti types of KTI mobility. The TIA values of Pannónia Kincse (9.8 ± 0.48 mg/g) were not significantly different (p ≤ 0.05) from Ti-a samples (10.07 ± 1.86 mg/g), while of Aires, Bahia, Hilario (6.19 ± 1.89) were identical (p ≤ 0.05) with ti samples (6.63 ± 1.99). Radiofrequency heat treatment (RF) decreased TIA values (p ≤ 0.05) at ≥ 100 °C. However, in the traditional soybean variety, the RF at 110 °C was more effective in eliminating the residual KTI activity. The remaining or the disapperaing bioactive form of trypsin inhibitors were succesfully characterized by the means of a standardized in vitro digestion model. It showed that residual BBI-originated trypsin inhibitor activity was in the stomach even after RF at 110 °C, whereas its chymotrypsin inhibitor activity was not detectable at all. Although PK and KTI null types of soybean seeds still required an energy-saving, gentle heat treatment to inactive the trypsin inhibitors before using them as food or feed, the physicochemical properties and processing quality of soybean products were protected, improved.
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Affiliation(s)
- Krisztina Takács
- Institute of Food Science and Technology, Department of Nutrition Science, Hungarian University of Agriculture and Life Sciences, Somlói Road 14-16, 1118 Budapest, Hungary
| | - Erika E. Szabó
- Institute of Food Science and Technology, Department of Nutrition Science, Hungarian University of Agriculture and Life Sciences, Somlói Road 14-16, 1118 Budapest, Hungary
| | - András Nagy
- Institute of Food Science and Technology, Department of Nutrition Science, Hungarian University of Agriculture and Life Sciences, Somlói Road 14-16, 1118 Budapest, Hungary
| | - Zsuzsanna Cserhalmi
- Institute of Food Science and Technology, Department of Nutrition Science, Hungarian University of Agriculture and Life Sciences, Somlói Road 14-16, 1118 Budapest, Hungary
| | - János Falusi
- Cereal Research Non-Profit Ltd, Alsókikötősor 9, 6726 Szeged, Hungary
| | - Éva Gelencsér
- Institute of Food Science and Technology, Department of Nutrition Science, Hungarian University of Agriculture and Life Sciences, Somlói Road 14-16, 1118 Budapest, Hungary
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Thompson HJ. Improving Human Dietary Choices Through Understanding of the Tolerance and Toxicity of Pulse Crop Constituents. Curr Opin Food Sci 2019; 30:93-97. [PMID: 32864345 PMCID: PMC7449238 DOI: 10.1016/j.cofs.2019.01.001] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/19/2023]
Abstract
Chickpea, dry bean, dry pea, and lentil are prominent dietary grain legumes commonly referred to as pulses. Pulses have been a staple component of the human diet for more than 8,000 years; however, in the last 70 years they have virtually disappeared from most Western diets. Reduced intake has occurred concomitantly with inadequate dietary fiber consumption and the onset of the obesity pandemic. Misinformation about tolerance and toxicity of several pulse crop constituents remains a barrier to public health efforts to increase dietary intake. Of particular concern are lectins which participate in agglutination reactions with cell surface proteins and galacto-oligosaccharides which have been associated with intestinal discomfort and flatulence. The scientific basis of these concerns is reviewed.
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Affiliation(s)
- Henry J Thompson
- Cancer Prevention Laboratory, Colorado State University, Fort Collins, CO 80523-1173
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Clemente A, Arques MDC. Bowman-Birk inhibitors from legumes as colorectal chemopreventive agents. World J Gastroenterol 2014; 20:10305-10315. [PMID: 25132747 PMCID: PMC4130838 DOI: 10.3748/wjg.v20.i30.10305] [Citation(s) in RCA: 65] [Impact Index Per Article: 5.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/13/2013] [Revised: 02/21/2014] [Accepted: 04/23/2014] [Indexed: 02/06/2023] Open
Abstract
Aberrant functioning of serine proteases in inflammatory and carcinogenic processes within the gastrointestinal tract (GIT) has prompted scientists to investigate the potential of serine protease inhibitors, both natural and synthetic, as modulators of their proteolytic activities. Protease inhibitors of the Bowman-Birk type, a major protease inhibitor family in legume seeds, which inhibit potently and specifically trypsin- and chymotrypsin-like proteases, are currently being investigated as colorectal chemopreventive agents. Physiologically relevant amounts of Bowman-Birk inhibitors (BBI) can reach the large intestine in active form due to their extraordinary resistance to extreme conditions within the GIT. Studies in animal models have proven that dietary BBI from several legume sources, including soybean, pea, lentil and chickpea, can prevent or suppress carcinogenic and inflammatory processes within the GIT. Although the therapeutic targets and the action mechanism of BBI have not yet been elucidated, the emerging evidence suggests that BBI exert their preventive properties via protease inhibition; in this sense, serine proteases should be considered as primary targets in early stages of carcinogenesis. The validation of candidate serine proteases as therapeutic targets together with the identification, within the wide array of natural BBI variants, of the most potent and specific protease inhibitors, are necessary to better understand the potential of this protein family as colorectal chemopreventive agents.
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Abstract
The Bowman-Birk inhibitor (BBI) is a small water-soluble protein present in soybean and almost all monocotyledonous and dicotyledonous seeds. The molecular size of BBI ranges from 1,513 Da to about 20,000 Da. BBI is to seeds what alpha(1)-antitrypsin is to humans. Soy-based food products rich in BBI include soybean grits, soymilk, oilcake, soybean isolate, and soybean protein concentrate. BBI is stable within the pH range encountered in most foods, can withstand boiling water temperature for 10 min, resistant to the pH range and proteolytic enzymes of the gastrointestinal tract, bioavailable, and not allergenic. BBI reduces the proteolytic activities of trypsin, chymotrypsin, elastase, cathepsin G, and chymase, serine protease-dependent matrix metalloproteinases, urokinase protein activator, mitogen activated protein kinase, and PI3 kinase, and upregulates connexin 43 (Cx43) expression. Several studies have demonstrated the efficacy of BBI against tumor cells in vitro, animal models, and human phase IIa clinical trials. FDA considers BBI as a drug. In 1999, FDA allowed a health claim on food labels stating that a daily diet containing 25 grams of soy protein, also low in saturated fat and cholesterol, may reduce the risk of heart disease [corrected] This review highlights the biochemical and functional food properties of the Bowman-Birk inhibitor.
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Affiliation(s)
- Jack N Losso
- Food Protein Biotechnology Laboratory, Department of Food Science, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA.
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Frokiær H, Mortensen K, Sorensen H, Sorensen S. Characterization of Proteintype Proteinase Inhibitors by High Performance Capillary Electrophoresis. J LIQ CHROMATOGR R T 2006. [DOI: 10.1080/10826079608006289] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/23/2022]
Affiliation(s)
- H. Frokiær
- a Department of Biochemistry and Nutrition Technical , University of Denmark , Building 224 DK-2800, Lyngby, Denmark
| | - K. Mortensen
- b Chemistry Department Royal Veterinary and Agricultural University , 40 Thorvaldsensvej DK-1871 Frederiksberg C, Denmark
| | - H. Sorensen
- b Chemistry Department Royal Veterinary and Agricultural University , 40 Thorvaldsensvej DK-1871 Frederiksberg C, Denmark
| | - S. Sorensen
- a Department of Biochemistry and Nutrition Technical , University of Denmark , Building 224 DK-2800, Lyngby, Denmark
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Devaraj VR, Manjunath NH. Effect of cooking on proteinase inhibitors of Dolichos lablab bean (Dolichos lablab perpureus L.). PLANT FOODS FOR HUMAN NUTRITION (DORDRECHT, NETHERLANDS) 1995; 48:107-112. [PMID: 8837868 DOI: 10.1007/bf01088305] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/22/2023]
Abstract
Proteinase inhibitory activity in ten different varieties of Dolichos lablab perpureus. L. was determined. All the varieties tested exhibited appreciable level of proteinase inhibitory activity (PIA). The trypsin inhibitory activity (TIA) (Mean: 20170 TIU/g) was relatively higher than the chymotrypsin inhibitory activity (CIA) (Mean: 15380 CIU/g). Effect of temperature and cooking on PIA was studied. The nature of cooking medium and duration of cooking had profound effect on the PIA. The dry fried seeds lost their PIA very rapidly (91% in 20 min). Seeds cooked in slightly alkaline medium lost their PIA quickly (89% in 30 min) compared to those cooked in acidic (80% in 30 min) and neutral pH (83% in 30 min). The PIA in green pods was also determined and they had only one third of the PIA (8200 TIU/g and 8125 CIU/g) found in the dry seeds.
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Affiliation(s)
- V R Devaraj
- Department of Chemistry, Bangalore University, India
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Weder JK, Haussner K, Bokor MV. Use of fluorogenic substrates to visualize trypsin and chymotrypsin inhibitors after electrophoresis. Electrophoresis 1993; 14:220-6. [PMID: 8486134 DOI: 10.1002/elps.1150140136] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/31/2023]
Abstract
Fluorogenic substrates were tested as a means of increasing both the sensitivity and the selectivity of trypsin and chymotrypsin inhibitor detection after electrophoretic separation. Out of six substrates applied to cellulose acetate membranes, N alpha-benzyloxycarbonyl-L-arginine-4-methylcoumarinyl-7-amide (Z-Arg-MCA) and benzyloxycarbonyl-glycyl-glycyl-L-arginine-4-trifluoromethylcoumariny l-7-amide (Z-Gly-Gly-Arg-TFMCA) were found to be suitable for trypsin, and L-alanyl-L-alanyl-L-phenylalanine-4-methylcoumarinyl-7-amide (Ala-Ala-Phe-MCA) was suitable for chymotrypsin. A procedure to detect trypsin and chymotrypsin inhibitors, and to discriminate between them, was developed. After electrophoresis, slab gels were first incubated with the enzyme (bovine trypsin, bovine chymotrypsin, or human duodenal juice) at 37 degrees C, and then covered with the respective substrate membrane and incubated at room temperature while being observed under UV light. Dark blue inhibitor bands on a light-blue-fluorescent background were obtained with Z-Arg-MCA/trypsin and Ala-Ala-Phe-MCA/chymotrypsin, whereas Z-Gly-Gly-Arg-TFMCA/trypsin resulted in dark inhibitor bands on a fluorescent green background. The "inhibitor overlay membrane technique" (IOM technique) was used after polyacrylamide gel isoelectric focusing with carrier ampholytes and immobilized pH gradients, pore-gradient polyacrylamide gel electrophoresis, and sodium dodecyl sulfate pore-gradient polyacrylamide gel electrophoresis.
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Affiliation(s)
- J K Weder
- Institut für Lebensmittelchemie, Technische Universität München, Garching, Germany
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Abstract
Perhaps with the notable exception, and that only in recent years, of red meat, which contributes dietary saturated fats and cholesterol, two well-known reasons in the etiology of heart-related disorders, no single group of foods has been portrayed in such negative terms as the food legumes traditionally have been during the last 50 years of research in food science and human nutrition. Even more alarming are the trends of continued research on such aspects as the deficiency of sulfur-amino acids (both by amino acid analyses as well as rat feeding studies), and the heat lability/stability of proteinase inhibitors and phytohemagglutinins in various legume species. A survey of literature indicates that over 100 research papers were published during the 1981 to 1990 period alone, in just three journals (Journal of Food Science, Journal of Agriculture and Food Chemistry, and Journal of the Science of Food and Agriculture) having the highest citation ratings in food sciences on these three topics, with a general consensus about the facts that were well established as early as the late 1950s. Considering the proliferation of journals publishing food science and human nutrition related work, especially in the Third World countries, the actual number probably would be much higher. This trend also indicates that we are repeating certain aspects of research on the importance of food legumes in human nutrition. Are we really any closer today in our understanding and appreciation of why the nomadic human made such a choice for their very existence during the transition to a more civilized society? This is a high time to project the image of legumes in human nutrition in proper perspective. The validity of our continued research on certain aspects of legumes in human nutrition, at a time when worldwide the research dollars are becoming increasingly harder to come by, is challenged in this review. Essentially, it is a journey through the author's personal diary that raises several questions in justifying the continued research support for at least some nutrition-related work on legumes and an account of what research areas perhaps need to be targeted in the 21st century.
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Affiliation(s)
- S S Deshpande
- Agriculture Canada Research Station, Morden, Manitoba
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Thorsen LI, Holm H, Reseland JE, Bjørnsen S, Hanssen LE, Brosstad F. Characterization of a human trypsin resistant to Kunitz soybean trypsin inhibitor. Studies of duodenal juices after tube instillation of raw soybean extract. Scand J Gastroenterol 1991; 26:589-98. [PMID: 1713706 DOI: 10.3109/00365529109043632] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/04/2023]
Abstract
Human duodenal juices collected during tube instillation of raw soybean extract into the duodenum contained free trypsin and free Kuntiz soybean trypsin inhibitor (KTI) in the simultaneous presence of trypsin-KTI complexes. It has previously been suggested that this KTI-non-inhibitable trypsin has a general resistance to serine protease inhibitors. Four different trypsin forms have been found and partly characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and isoelectric focusing followed by Western immunoblotting or enzyme staining. In addition, crossed immunoelectrophoresis and affinity chromatography with antibody-coupled gels have been used for identification of free and inhibitor-complexed trypsin.
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Affiliation(s)
- L I Thorsen
- Matforsk, Norwegian Food Research Institute, Aas
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Weder JK, Haussner K. Inhibitors of human and bovine trypsin and chymotrypsin in fenugreek (Trigonella foenum-graecum L.) seeds. Isolation and characterization. ZEITSCHRIFT FUR LEBENSMITTEL-UNTERSUCHUNG UND -FORSCHUNG 1991; 192:535-40. [PMID: 1872034 DOI: 10.1007/bf01202509] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/29/2022]
Abstract
Three fenugreek inhibitors (TFI-A8, TFI-N2, and TFI-B2) were isolated from an inhibitor preparation by anion exchange chromatography and subsequent preparative isoelectric focusing using immobilized pH gradients and the canal technique. The purified inhibitors inhibited the enzymes tested differently: TFI-A8 exhibited a high inhibition of trypsin (8.2 mg human trypsin/mg and 8.1 mg bovine trypsin/mg) and a very low inhibition of chymotrypsin (0.8 mg human chymotrypsin/mg and 1.0 mg bovine chymotrypsin/mg). TFI-N2 inhibited the four enzymes to about the same extent (5.0 mg/mg human and 4.1 mg/mg bovine trypsin; 4.9 mg/mg human and 3.7 mg/mg bovine chymotrypsin). TFI-B2 displayed a high inhibition of trypsin (7.5 mg/mg human and 5.1 mg/mg bovine) and a low inhibition of chymotrypsin (1.8 mg/mg human and 1.9 mg/mg bovine). On average, the human enzymes were inhibited better than the bovine ones by the purified inhibitors. The inhibitors contained high amounts of cystine (five or six disulfide bridges per molecule), aspartic acid, threonine, serine and proline, no valine and methionine and two of them also no tryptophan. Their molecular masses were about 6 kDa. Their inclusion into the Bowman-Birk soybean proteinase inhibitor family is discussed.
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Affiliation(s)
- J K Weder
- Institut für Lebensmittelchemie, Technische Universität München, Federal Republic of Germany
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Weder JK, Haussner K. Inhibitors of human and bovine trypsin and chymotrypsin in fenugreek (Trigonella foenum-graecum L.) seeds. Demonstration and purification. ZEITSCHRIFT FUR LEBENSMITTEL-UNTERSUCHUNG UND -FORSCHUNG 1991; 192:455-9. [PMID: 2058314 DOI: 10.1007/bf01193147] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/30/2022]
Abstract
Fenugreek contained proteinase inhibitors inhibiting 5-9 mg human trypsin, 5-7 mg bovine trypsin, 2-6 mg human chymotrypsin, and 1-3 mg bovine chymotrypsin per g seed material. About 30 inhibitors were electrophoretically detected, and 23 of them, inhibiting all the four enzymes, were characterized by means of their isoelectric points: a group of acid inhibitors (TFI-A1 to A10, pI 4.48-5.12), a group of neutral inhibitors (TFI-N1 to -N6, pI 5.91-6.71), and a group of basic inhibitors (TFI-B1 to -B7, pI 7.76-9.77). To eliminate the galactomannans which complicate further purification, coarsely ground seeds were separated by density into two fractions, seed coats + endosperm and cotyledons + embryos (C + E). Isolation of the fenugreek inhibitors by extraction of fraction C + E, followed by ammonium sulfate fractionation and affinity chromatography on anhydrotrypsin-Sepharose, resulted in an about 700-fold enrichment.
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Affiliation(s)
- J K Weder
- Institut für Lebensmittelchemie, Technische Universität München, Garching, Federal Republic of Germany
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WEDER JUERGENKP, MUELLER RUDI. REACTION OF LENTIL PROTEINASE INHIBITORS WITH HUMAN AND BOVINE TRYPSIN AND CHYMOTRYPSIN. J Food Biochem 1989. [DOI: 10.1111/j.1745-4514.1989.tb00387.x] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/28/2022]
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MUELLER RUDI, WEDER JUERGENKP. ISOLATION AND CHARACTERIZATION OF TWO TRYPSIN-CHYMOTRYPSIN INHIBITORS FROM LENTIL SEEDS (LENS CULINARIS MEDIK.). J Food Biochem 1989. [DOI: 10.1111/j.1745-4514.1989.tb00384.x] [Citation(s) in RCA: 27] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/27/2022]
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