MOLECULAR PROFILE, PURITY AND PRESENCE OF TRYPSIN INHIBITORS IN COWPEA PROTEIN ISOLATES
DOI:
https://doi.org/10.1590/1983-21252018v31n123rcKeywords:
Vigna unguiculata. Trypsin inhibitor. Protein isolate.Abstract
The most-used preparation process of protein isolates (PI) involves the isoelectric precipitation of the protein. Heating shortens the preparation time but this procedure may affect the purity, yield, molecular profile of the protein, and the activity of the trypsin inhibitor. This study aimed to investigate the effect of heating in the production of cowpea protein isolates. Crude whole beans (WB) were defatted with hexane, and the protein isolates obtained by isoelectric precipitation with (HPI), and without (NHPI) heating. The protein content of the WB and the PI was determined by the micro-Kjeldahl method, and the extraction yield estimated from the protein content at the end of extraction in relation to this content in the raw material. Possible losses of protein fractions were followed by SDS-PAGE, and the trypsin inhibitor activity determined by an enzymatic assay (BAPNA: benzoyl-DL-arginine-p-nitroanilide). Protein content in HPI was 83.3%, less than in the NHPI (92.2%). The HPI yield was lower (40.0%) as compared to the NHPI (42.3%). Electrophoresis indicated bands ranging from 13 to 262 kDa in WB; and the NHPI presented a protein fraction’s profile closer to that of the WB than to the HPI. The WB had the trypsin inhibitor activity, expressed as Trypsin Inhibitory Units (TIU), of 32.5±0.5 TIU /mg-protein; HPI showed 12.7±0.5 TIU /mg-protein (39% of that observed in WB) and the NHPI, 8.3±0.2 TIU /mg-protein (25.5%). Heating reduces the yield and purity of proteins in the isolates. However, the inhibitory activity of trypsin cowpea is most affected by the isolation procedure.Downloads
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References
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WANG, M. Optimization of extraction process of protein isolate from mung bean Procedia Engineering, Amsterdam, v. 15, n. 1, p. 5250-5258, 2011.
WITHANA-GAMAGE, T. S. et al. Physicochemical. thermal and functional characterization of protein isolates from Kabuli and Desi chickpea (Cicer arietinum L.): a comparative study with soy (Glycine max) and pea (Pisum sativum L.). Journal of the Science of Food and Agriculture, New Jersey, v. 91, n. 6, p. 1022-1031, 2011.
WRIGHT, D. J.; BUMSTEAD, M. R. Legume proteins in food technology. Philosophical Transactions of the Royal Society of London, London, v. 304, n. 1120, p. 381-393, 1984.
BOYE, J.; ZARE, F.; PLETCH, A. Molecular. functional and processing characteristics of whole pulses and pulse fractions and their emerging food and nutraceutical applications. Food Research International, Amsterdam, v. 43, n. 2, p. 414-431, 2010.
BUTT, M. S.; BATOOL, R. Nutritional and functional properties of some promising legumes protein isolates. Pakistan Journal of Nutrition, Dubai, v. 9, n. 4, p. 373-379, 2010.
DEAK, N. A.; JOHNSON, L. A. Effects of extraction temperature and preservation method on functionality of soy protein. Journal of the American Oil Chemists Society, Chicago, v. 84, n. 3, p. 259-268, 2007.
DUARTE, M. S. L. et al. Determination of the trypsin inhibitor activity in vitro of the black bean (Phaseolus vulgaris L.) albumin and globulin. Alimentos e Nutrição, Araraquara, v. 21, n. 3, p. 373-376, 2010.
ELHARDALLOU, S. et al. Amino acid composition of cowpea (Vigna ungiculata L. Walp.) flour and its protein isolates. Food and Nutrition Sciences, Irvine, v. 6, n. 9, p. 790-797, 2015.
FRIEDMAN, M.; BRANDON, D. L. Nutritional and health benefits of soy proteins. Journal of Agricultural and Food Chemistry, Washington, v. 49, n. 3, p. 1069-1086, 2001.
FROTA, K. M. G. et al. Cholesterol-lowering properties of whole cowpea seed and its protein isolate in hamsters. Journal of Food Science. New Jersey, v. 73, n. 9, p. 235-240, 2008.
FROTA, K. M. et al. Cowpea protein reduces LDL-cholesterol and apolipoprotein B concentrations. but does not improve biomarkers of inflammation or endothelial dysfunction in adults with moderate hypercholesterolemia. Nutrición Hospitalaria, Barcelona, v. 31, n. 4, p. 1611-1619, 2015.
FROTA, K. M. G.; SOARES, R. A. M.; ARÊAS, J. A. G. Composição química do feijão caupi (Vigna unguiculata L. Walp), cultivar BRS-Milênio. Ciência e Tecnologia de Alimentos, Campinas, v. 28, n. 2, p. 470-476, 2008.
HONIG, D. H.; RACKIS, J. J.; WOLF, W. J. Effects of pH and salt on yields. trypsin inhibitor contends. and mineral levels of soybean protein isolate and whey. Journal of Agricultural and Food Chemistry, Washington, v. 35, n. 6, p. 967-971, 1987.
HORAX, R. et al. Functional properties of protein isolate from cowpea (Vigna unguiculata L. Walp.). Journal of Food Science, Oxford, v. 69, n. 2, p. 119–121, 2004.
JOURDAN, G. A.; NOREÑA, C. P. Z.; BRANDELLI, A. Inactivation of trypsin inhibitor activity from Brazilian varieties of beans (Phaseolus vulgaris L.). Food Science and Technology International, London, v. 13, n. 3, p. 195–198, 2007.
KAKADE, M. L.; SIMONS, N.; LIENER, I. E. An evaluation of natural vs synthetic substrates for measuring the antitryptic activity of soybean samples. Cereal Chemistry, Sydney, v. 46, n. 5, p. 518-526, 1969.
KALPANADEVI, V.; MOHAN, V. R. Effect of processing on antinutrients and in vitro protein digestibility of the underutilized legume. Vigna unguiculata (L.) Walp. subsp. unguiculata. LWT-Food Science and Technology, Amsterdam, v. 51, n. 2, p. 455-461, 2013
LAEMMLI, U. K. Cleavage of structural proteins during assembly of the heat of bacteriophage T4. Nature, London, v. 227, n. 5259, p. 680-685, 1970.
LAJOLO, F. M.; GENOVESE, M. Nutritional significance of lectins and enzyme inhibitors from legumes. Journal of Agricultural and Food Chemistry, Washington, v. 50, n. 22, p. 6592-6598, 2002.
LIENER, I. E. Implications of antinutritional components in soybean foods. Critical Reviews in Food Science and Nutrition, Philadelphia, v. 34, n. 1, p. 31-67, 1994.
MARQUES, M. R. et al. Proteolytic hydrolysis of cowpea proteins is able to release peptides with hypocholesterolemic activity. Food Research International, Amsterdam, v. 77, n. 1, p. 43-49, 2015.
PEYRANO, F.; SPERONI, F.; AVANZA, M. V. Physicochemical and functional properties of cowpea protein isolates treated with temperature or high hydrostatic pressure. Innovative Food Science and Emerging Technologies, Amsterdam, v. 33, n. 1, p. 38-46, 2016.
RANGEL, A. et al. Biological evaluation of a protein isolate from cowpea (Vigna unguiculata) seeds. Food Chemistry, Washington, v. 87, n. 4, p. 491-499, 2004.
RAYAS-DUARTE, P.; BERGERON, D.; NIELSEN, S. Screening of heat-stable trypsin inhibitors in dry beans and their partial purification from great Northern beans (Phaseolus vulgaris) using anhydrotrypsin-sepharose affinity chromatography. Journal of Agricultural and Food Chemistry, Washington, v. 40, n. 1, p. 32-42, 1992.
RIVAS-VEGA, M. E. et al. Nutritional value of cowpea (Vigna unguiculata L. Walp.) meals as ingredients in diets for Pacific white shrimp (Litopenaeus vannamei Boone). Food Chemistry, Washington, v. 97, n. 1, p. 41-49, 2006.
RUSSO, R.; REGGIANI, R. Evaluation of protein concentration. amino acid profile and antinutritional compounds in hempseed meal from dioecious and monoecious varieties. American Journal of Plant Sciences, Irvine, v. 6, n. 1, p. 14-22, 2015.
SARWAR, G.; XIAO, C.; COCKELL, K. A. Impact of antinutritional factors in food proteins on the digestibility of protein and the bioavailability of amino acids and on protein quality. The British Journal of Nutrition, London, v. 108, n. 2, p. 315-332, 2012.
SING, B. B. et al. Cowpea genetics: a review of the recent literature. In: Advances in cowpea research. Ibadan: IITA-JIRCAS. 1997. cap. 2. p. 13-15.
UZZAN, A. Vegetable protein products from seeds: technology and uses in food industry. In: HUDSON. B. J. F. (Ed.). Development in Food Proteins. New York: Elsevier Applied Science. 1988. p. 73-118.
WAGNER, J. R.; SORGENTINI, D. A.; ANÕN, M. C. Relation between Solubility and Surface Hydrophobicity as an Indicator of Modifications during Preparation Processes of Commercial and Laboratory-Prepared Soy Protein Isolates. Journal of Agricultural and Food Chemistry, Washington, v. 48, n. 8, p. 3159-3165, 2000.
WANG, M. Optimization of extraction process of protein isolate from mung bean Procedia Engineering, Amsterdam, v. 15, n. 1, p. 5250-5258, 2011.
WITHANA-GAMAGE, T. S. et al. Physicochemical. thermal and functional characterization of protein isolates from Kabuli and Desi chickpea (Cicer arietinum L.): a comparative study with soy (Glycine max) and pea (Pisum sativum L.). Journal of the Science of Food and Agriculture, New Jersey, v. 91, n. 6, p. 1022-1031, 2011.
WRIGHT, D. J.; BUMSTEAD, M. R. Legume proteins in food technology. Philosophical Transactions of the Royal Society of London, London, v. 304, n. 1120, p. 381-393, 1984.
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