FNS  Vol.3 No.4 , April 2012
Lima Bean (Phaseolus lunatus) Protein Hydrolysates with ACE-I Inhibitory Activity
ABSTRACT
Several protein sources can be used to produce bioactive peptides with angiotensin I-converting enzyme (ACE) inhibittory activity. Protein concentrates from ungerminated and germinated lima bean Phaseolus lunatus seed flours were hydrolyzed with Alcalase 2.4 L or pepsin-pancreatin sequential hydrolysis, and ACE inhibitory activity measured in the different hydrolysis treatments. Protein hydrolysate production was analyzed with a 23 factorial design with four replicates of the central treatment. Evaluated factors were protein concentrate source (ungerminated seeds, PC1; germinated seeds, PC2), enzyme/substrate ratio E/S (1/50 or 1/10) and hydrolysis time (0.5 or 2.0 h for Alcalase; 1 or 3 h for pepsin-pancreatin). Degree of hydrolysis (DH) was high for the Alcalase hydrolysates (24.12% 58.94%), but the pepsin-pancreatin hydrolysates exhibited the highest ACE inhibitory activity (IC50 = 0.250 0.692 mg/mL). Under the tested conditions, the hydrolysates with the highest ACE inhibitory activity were produced with sequential pepsin-pancreatin using either PC1 at 1 h hydrolysis time and a 1/10 E/S ratio or PC2 at 1 h hydrolysis time and a 1/50 E/S ratio. Lima bean protein hydrolysates prepared with Alcalase or pepsin-pancreatin are a potential ingredient in the production of physiologically functional foods with antihypertensive activity.

Cite this paper
L. Chel-Guerrero, M. Domínguez-Magaña, A. Martínez-Ayala, G. Dávila-Ortiz and D. Betancur-Ancona, "Lima Bean (Phaseolus lunatus) Protein Hydrolysates with ACE-I Inhibitory Activity," Food and Nutrition Sciences, Vol. 3 No. 4, 2012, pp. 511-521. doi: 10.4236/fns.2012.34072.
References
[1]   K. S. Mark and T. P. Davis, “Stroke: Development, Prevention and Treatment with Peptidase Inhibitors,” Peptides, Vol. 21, No. 12, 2000, pp. 1965-1973. doi:10.1016/S0196-9781(00)00346-6

[2]   R. Varin, P. Mulder, F. Tamion, V. Richard, J. P. Henry, F. Lallemand, G. Lerebours and C. Thuillez, “Improvement of Endothelial Function by Chronic Angiotensin Converting Enzyme Inhibition in Heart Failure: Role of Nitric Oxide, Prostanoids, Oxidant Stress, and Bradykinin,” Circulation, Vol. 102, 2000, pp. 351-356.

[3]   A. F. Lever, D. J. Hole, C. R. Gillis, I. R. McCallum, G. T. McInnes, P. L. McKinnon, P. A. Meredith, L. S. Murray, J. L. Reid and J. W. Robertson, “Do Inhibitors of Angiotensin-I-Converting Enzyme Protect against Risk of Cancer?” The Lancet, Vol. 352, No. 9123, 1998, pp. 179184. doi:10.1016/S0140-6736(98)03228-0

[4]   A. F. Lever, D. J. Hole, C. R. Gillis, G. T. McInnes, P. A. Meredith, L. S. Murray and J. L. Reid, “Is Cancer Related to Hypertension or to Its Treatment?” Clinical Experimental Hypertension, Vol. 21, 1999, pp. 937-946. doi:10.3109/10641969909061022

[5]   H. Gavras and I. Gavras, “Bioactive Peptides in the Treatment of Hypertension and Heart Failure,” Biomedical and Health Research, Vol. 22, 1999, pp. 41-50.

[6]   G. H. Li, Y. H. Shi, H. Liu and G. W. Le, “Antihypertensive Effect of Alcalase Generated Mung Bean Protein Hydrolysates in Spontaneously Hypertensive Rats,” European Food Research and Technology, Vol. 222, No. 5-6, 2006, pp. 733-736. doi:10.1007/s00217-005-0147-2

[7]   G. H. Li, G. W. Le, Y. H. Shi and Y. Shrestha, “Angiotensin I-Converting Enzyme Inhibitory Peptides Derived From Food Proteins and Their Physiological and Pharmacological Effects,” Nutrition Research, Vol. 24, 2004, pp. 469-486.

[8]   A. Quirós, R. Chichón, I. Recio and R. López-Fandi?o, “The Use of High Hydrostatic Pressure to Promote the Proteolysis and Release of Bioactive Peptides from Ovalbumin,” Food Chemistry, Vol. 54, 2006, pp. 19-27.

[9]   M. Miguel, M. J. Alonso, M. Salaices, A. Aleixandre and R. López-Fandi?o, “Antihypertensive, ACE-inhibitory and Vasodilator Properties of an Egg White Hydrolysate: Effect of a Simulated Intestinal Digestion,” Food Chemistry, Vol. 104, No. 1, 2007, pp. 163-168. doi:10.1016/j.foodchem.2006.11.016

[10]   G. H. Li, G. W. Le, H. Liu and Y. H. Shi, “Mung-bean Protein Hydrolysates Obtained with Alcalase Exhibit Angiotensin I-Converting Enzyme Inhibitory Activity,” Food Science and Technology International, Vol. 11, No. 4, 2005, pp. 281-287. doi:10.1177/1082013205056781

[11]   Z. I. Shin, C. W. Ahn, H. S. Nam, H. J. Lee and T. H. Moon, “Fractionation of Angiotensin Converting Enzyme Inhibitory Peptide from Soybean Paste,” Korean Journal of Food Science and Technology, Vol. 27, 1995, pp. 230-234.

[12]   Z. I. Shin, R. Yu, S. A. Park, D. K. Chung, H. S. Nam, K. S. Kim and H. J. Lee. “His-His-Leu, an Angiotensin I Converting Enzyme Inhibitory Peptide Derived from Korean Soybean Paste, Exerts Antihypertensive Activity in Vivo,” Journal of Agricultural and Food Chemistry, Vol. 49, No. 6, 2001, pp. 3004-3009. doi:10.1021/jf001135r

[13]   V. Vermeirssen, C. P. Van, L. Devos and W. Verstraete, “Release of Angiotensin I Converting Enzyme (ACE). Inhibitory Activity during in Vitro Gastrointestinal Digestion: From Batch Experiment to Semicontinuous Model,” Journal of Agriculture and Food Chemistry, Vol. 51, No. 19, 2003, pp. 5680-5687. doi:10.1021/jf034097v

[14]   M. M. Yust, J. Pedroche, J. Girón-Calle, M. Alaiz, F. Millán and J. Vioque, “Production of ACE Inhibitory Peptides by Digestion of Chickpea Legumin with Alcalase,” Food Chemistry, Vol. 81, No. 3, 2003, pp. 363-369. doi:10.1016/S0308-8146(02)00431-4

[15]   J. Pedroche, M. M. Yust, C. Megías, H. Lqari, M. Alaiz, J. Girón-Calle, F. Millán and J. Vioque, “Utilization of Rapeseed Protein Isolates for Production of Peptides with Angiotensin I-Converting Enzyme (ACE)-Inhibitory Activity,” Grasas y Aceites, Vol. 55, 2004, pp. 354-358.

[16]   Y. Yoshie-Stark, Y. Wada, M. Schott and A. W?she, “Functional and Bioactive Properties of Rapeseed Protein Concentrates and Sensory Analysis of Food Application with Rapeseed Protein Concentrates,” LWT—Food Science and Technology, Vol. 39, No. 5, 2006, pp. 503-512. doi:10.1016/j.lwt.2005.03.006

[17]   A. Manninen, “Protein Hydrolysates in Sports and Exercise: A Brief Review,” Journal of Sports Science and Medicine, Vol. 3, 2004, pp. 60-63.

[18]   H. G. Kristinsson and B. A. Rasco, “Biochemical and Functional Properties of Atlantic Salmon (Salmo salar) Muscle Proteins Hydrolyzed with Various Alkaline Proteases,” Journal of Agricultural and Food Chemistry, Vol. 48, No. 3, 2000, pp. 657-666. doi:10.1021/jf990447v

[19]   D. Doucet, D. E. Otter, S. F. Gauthier and E. A. Foegeding, “Enzyme-induced Gelation of Extensively Hydrolyzed Whey Proteins by Alcalase: Peptide Identification and Determination of Enzyme Specificity,” Journal of Agricultural and Food Chemistry, Vol. 51, No. 21, 2003, pp. 6300-6308. doi:10.1021/jf026242v

[20]   P. W. Tardioli, R. Fernández-Lafuente, J. M. Guisán and R. L. C. Giordano, “Design of New Immobilized-Sabilized Carboxypeptidase a Derivative for Production of Aromatic Free Hydrolysates of Proteins,” Biotechnology Progress, Vol. 19, No. 2, 2003, pp. 565-574. doi:10.1021/bp0256364

[21]   L. Chel-Guerrero, V. Pérez-Flores, D. Betancur-Ancona and G. Dávila-Ortíz, “Functional Properties of Flours and Protein Isolates from Phaseolus lunatus and Canavalia ensiformis Seeds,” Journal of Agricultural and Food Chemistry, Vol. 5, 2002, pp. 584-591. doi:10.1021/jf010778j

[22]   N. Lecomte, J. Zayas and C. Kastner, “Soy Proteins Functional and Sensory Characteristics Improved in Comminuted Meals,” Journal of Food Science, Vol. 58, No. 3, 1993, pp. 464-466. doi:10.1111/j.1365-2621.1993.tb04300.x

[23]   Y. H. Kuo, P. Rosan, F. Lambein, J. Frias and C. Vidal-Valverde, “Effects of Different Germination Conditions on the Contents of Free Protein and Non-protein Amino Acids of Commercial Legumes,” Journal of Food Chemistry, Vol. 86, No. 4, 2004, pp. 537-545. doi:10.1016/j.foodchem.2003.09.042

[24]   A. A. Oshodi and V. A. Aletor, “Functional Properties of Haemagglutinins (Lectins) Extracted from Some Edible Varieties of Lima Beans (Phaseolus lunatus L),” International Journal of Food Science and Nutrition, Vol. 44, 1993, pp. 133-136. doi:10.3109/09637489309017431

[25]   R. Sanchez-Vioque, A. Clemente, J. Vioque, J. Bautista and F. Millan, “Protein Isolates from Chickpea (Cicer arietinum L.): Chemical Composition, Functional Properties and Protein Characterization,” Food Chemistry, Vol. 64, No. 2, 1999, pp. 237-243. doi:10.1016/S0308-8146(98)00133-2

[26]   Anonymous, “International Rules for Seeds Testing,” The International Seed Testing Association (ISTA), Switzerland, 1999.

[27]   A.O.A.C, “Official Methods of Analysis of the Association of Analytical Chemists,” 15th Edition, National Academy Press, Washington DC, 1995.

[28]   M. Alaiz, J. L. Navarro, J. Giron and J. Vioque, “Amino Acid Analysis by High Performance Liquid Chromatography after Derivatization with Diethyletoxymethylenemalonate,” Journal of Chromatography, Vol. 591, No. 1-2, 1992, pp. 181-186. doi:10.1016/0021-9673(92)80236-N

[29]   C. Megías, M. M. Yust, J. Pedroche, H. Lquari, J. Girón-Calle, M. Alaiz, F. Millán and J. Vioque, “Purification of an ACE Inhibitory Peptide Alter Hydrolysis of Sunflower (Helianthus annuus L.) Protein Isolates,” Journal of Agriculture and Food Chemistry, Vol. 52, No. 7, 2004, pp. 1928-1932. doi:10.1021/jf034707r

[30]   P. M. Nielsen, D. Petersen and C. Dambmann, “Improved Method for Determining Food Protein Degree of Hydrolysis,” Journal of Food Science, Vol. 66, No. 5, 2001, pp. 642-646. doi:10.1111/j.1365-2621.2001.tb04614.x

[31]   M. Hayakari, Y. Kondo and H. Izumi, “A Rapid and Simple Spectrophotometric Assay of Angiotensin-Converting Enzyme,” Analytical Biochemistry, Vol. 84, No. 2, 1978, pp. 361-369. doi:10.1016/0003-2697(78)90053-2

[32]   D. C. Montgomery, “Dise?o y Análisis de Experimentos,” Editorial Limusa-Wiley, Segunda Edición, 2004, pp. 228-241.

[33]   A. N. Dibofori, P. N. Okoh and A. O. Onigbinde, “Effect of Germination on the Cyanide and Oligosaccharide Content of Lima Beans (Phaseolus lunatus),” Journal of Food Chemistry, Vol. 51, No. 2, 1994, pp. 133-136. doi:10.1016/0308-8146(94)90246-1

[34]   C. Rodríguez, J. Frias, C. Vidal-Valverde and A. Hernández, “Correlation between Some Nitrogen Fractions, Lysine, Histidine, Tyrosine, and Ornithine Contents During Germination of Peas, Beans, and Lentils,” Journal of Food Chemistry, Vol. 108, No. 1, 2008, pp. 245-252. doi:10.1016/j.foodchem.2007.10.073

[35]   H. Bau, C. Villaume, J. Nicolas and L. Méjean, “Effect of Germination on Chemical Composition, Biochemical Constituents and Antinutritional Factors of Soya Bean (Glycine max) Seeds,” Journal of Science Food and Agriculture, Vol. 73, No. 1, 1997, pp. 1-9. doi:10.1002/(SICI)1097-0010(199701)73:1<1::AID-JSFA694>3.0.CO;2-B

[36]   E. Sangronis, M. Rodríguez, R. Cava and A. Torres, “Protein Qality of Germinated Phaseolus vulgaris,” European Food Research and Technology, Vol. 222, No. 1-2, 2006, pp. 144-148. doi:10.1007/s00217-005-0137-4

[37]   J. Augustin and P. Klein, “Nutrient Composition of Raw, Cooked, Canned and Sprouted Legumes,” Matthews and Dekker, New York, 1989, pp. 187-204.

[38]   D. Betancur-Ancona, R. Martinez-Rosado, A. CoronaCruz, A. Castellanos-Ruelas, M. E. Jaramillo-Flores and L. Chel-Guerrero, “Functional Properties of Hydrolysates From Phaseolus lunatus Seeds,” International Journal of Food Science and Technology, Vol. 44, No. 1, 2009, pp. 128-137. doi:10.1111/j.1365-2621.2007.01690.x

[39]   D. Betancur-Ancona, S. Gallegos-Tintoré and L. ChelGuerrero, “Wet-fractionation of Phaseolus lunatus Seeds: Partial Characterization of Starch and Protein,” Journal of Science and Food and Agriculture, Vol. 84, No. 10, 2004, pp. 1193-1201. doi:10.1002/jsfa.1804

[40]   S. D. Arntfield, M. A. H. Ismond and E. D. Murray, “The Fate of Antinutritional Factors During the Preparation of Faba Bean Protein Isolate Using a Micellization Technique,” Canadian Institute of Food Science Technology Journal, Vol. 18, 1985, pp. 137-143.

[41]   S. L. Rodríguez-Ambriz, A. L. Martínez-Ayala, F. Millán and G. Dávila-Ortiz, “Composition and Functional Properties of Lupinus campestris Protein Isolates,” Plant Foods for Human Nutrition, Vol. 60, No. 3, 2005, pp. 99107. doi:10.1007/s11130-005-6835-z

[42]   O. Paredes-López and C. Ondorica-Falomir, “Production of Safflower Protein Isolates: Composition, Yield and Protein Quality,” Journal of the Science and Food Agriculture, Vol. 37, 1986, pp. 1104-1109.

[43]   H. G. Byun and S. K. Kim, “Purification and Characterization of Angiotensin I Converting Enzyme (ACE) Inhibitory Peptides from Alaska Pollack (Theragra chalcogramma) Skin,” Journal of Process Biochemistry, Vol. 36, No. 12, 2001, pp. 1155-1162. doi:10.1016/S0032-9592(00)00297-1

[44]   J. Pedroche, M. M. Yust, J. Girón-Calle, M. Alaiz, F. Millán and J. Vioque, “Utilization of Chickpea Protein Isolates for Production of Peptides with Angiotensin Iconverting Enzyme (ACE)-Inhibitory Activity,” Journal of the Science and Food Agriculture, Vol. 82, No. 9, 2002, pp. 960-965. doi:10.1002/jsfa.1126

[45]   I. M. Mahmoud, T. W. Malone and T. C. Cordle, “Enzymatic Hydrolysis of Casein: Effect of Degree of Hydrolysis on Antigenicity and Physical Properties,” Journal of Food Science, Vol. 57, No. 5, 1992, pp. 1223-1229. doi:10.1111/j.1365-2621.1992.tb11304.x

[46]   N. J. Adamson and E. C. Reynolds, “Characterization of Multiply Phosphorylated Peptides Selectively Precipitated from Pancreatin Casein Digest,” Journal of Dairy Science, Vol. 78, No. 12, 1995, pp. 2653-2659. doi:10.3168/jds.S0022-0302(95)76895-3

[47]   J. M. Kim, J. H. Whang and H. J. Suh, “Enhancement of Angiotensin I Convering Enzyme Inhibitory Activity and Improvement of the Emulsifying and Foaming Properties of Corn Gluten Hydrolysate Using Ultrafiltration Membranes,” European Food Research and Technology, Vol. 218, No. 2, 2004, pp. 133-138. doi:10.1007/s00217-003-0825-x

[48]   V. Vermeirssen, J. Van Camp and W. Verstraete, “Optimization and Validation of an Angiotensin-Converting Enzyme Inhibition Assay for Screening of Bioactive Peptides,” Journal of Biochemical and Biophysical Methods, Vol. 51, No. 1, 2002; pp. 75-87. doi:10.1016/S0165-022X(02)00006-4

[49]   C. H. Li, T. Matsui, K. Matsumoto, R. Yamasaki and T. Kawasaki, “Latent Production of Angiotensin I-Converting Enzyme Inhibitors from Buckwheat Protein,” Journal of Peptide Science Vol. 8, No. 6, 2002, pp. 267274. doi:10.1002/psc.387

[50]   M. Gobbetti, P. Ferranti, E. Smacchi, F. Goffredi and F. Addeo, “Production of Angiotensin-I-Converting-Enzyme-Inhibitory Peptides in Fermented Milks Started by Lactobacillus delbrueckii subsp. Bulgaricus SS1 and Lactococcus lactis subsp. Cremoris FT4,” Applied and Environmental Microbiology, Vol. 66, No. 9, 2000, pp. 3898-3904. doi:10.1128/AEM.66.9.3898-3904.2000

[51]   M. Miguel, M. J. Alonso, M. Salaices, A. Aleixandre and R. López-Fandi?o, “Antihipertensive, ACE-Inhibitory and Vasodilator Properties of an Egg White Hydrolysate: Effect of a Simulated Intestinal Digestion,” Food Chemistry, Vol. 104, No. 1, 2007, pp. 163-168. doi:10.1016/j.foodchem.2006.11.016

 
 
Top