Synthetic angiotensin I-converting enzyme (ACE-I) inhibitors can have undesirable side effects, while natural inhibitors have no side effects and are potential nutraceuticals. A protein-rich fraction from chia (Salvia hispanica L.) seed was hydrolyzed with an Alcalase-Flavourzyme sequential system and the hydrolysate ultrafiltered through four molecular weight cut-off membranes (1？kDa, 3？kDa, 5？kDa, and 10？kDa). ACE-I inhibitory activity was quantified in the hydrolysate and ultrafiltered fractions. The hydrolysate was extensive (DH = 51.64%) and had 58.46% ACE-inhibitory activity. Inhibition ranged from 53.84% to 69.31% in the five ultrafiltered fractions and was highest in the <1？kDa fraction (69.31%). This fraction’s amino acid composition was identified and then it was purified by gel filtration chromatography and ACE-I inhibition measured in the purified fractions. Amino acid composition suggested that hydrophobic residues contributed substantially to chia peptide ACE-I inhibitory strength, probably by blocking angiotensin II production. Inhibitory activity ranged from 48.41% to 62.58% in the purified fractions, but fraction F1 (1.5–2.5？kDa) exhibited the highest inhibition (IC50 = 3.97？μg/mL; 427–455？mL elution volume). The results point out the possibility of obtaining bioactive peptides from chia proteins by means of a controlled protein hydrolysis using Alcalase-Flavourzyme sequentional system. 1. Introduction Cardiovascular disease (CVD) affects the heart and blood vessels and is the principal cause of death worldwide. Considered the primary risk factor for CVD, high blood pressure, or hypertension, consists of a sustained increase in blood pressure levels. In 2000, >25% of the population worldwide (approximately 1 billion) suffered from hypertension, a figure predicted to increase to 1.56 billion by 2025 [1]. Angiotensin I-converting enzyme (peptidyl carboxy peptidase, EC 3.4.15.1, ACE) belongs to the class of zinc proteases that require zinc and chloride for activation. ACE plays an important role in blood pressure regulation via the renin-angiotensin system (RAS) and the kallikrein-kinnin system (KKS). In the KKS, ACE inactivates the vasodilator bradykinin, while in the RAS, ACE acts as an exopeptidase cleaving His-Leu from the C-terminal of decapeptide angiotensin I and producing the potent vasoconstrictor octapeptide angiotensin II [2]. Use of enzyme technologies for protein recovery and modification has led to production of a broad spectrum of food ingredients and industrial products. Hydrolysis selectivity is commonly manipulated by
References
[1]
C. L. Jao, S. L. Huang, and K. C. Hsu, “Angiotensin I-converting enzyme inhibitory peptides: inhibition mode, bioavailability, and antihypertensive effects,” BioMedicine, vol. 2, no. 4, pp. 130–136, 2012.
[2]
S. C. Ko, N. Kang, E. A. Kim et al., “A novel angiotensin I-converting enzyme (ACE) inhibitory peptide from a marine Chlorella ellipsoidea and its antihypertensive effect in spontaneously hypertensive rats,” Process Biochemistry, vol. 47, pp. 2005–2011, 2012.
[3]
S. Nalinanon, S. Benjakul, H. Kishimura, and F. Shahidi, “Functionalities and antioxidant properties of protein hydrolysates from the muscle of ornate threadfin bream treated with pepsin from skipjack tuna,” Food Chemistry, vol. 124, no. 4, pp. 1354–1362, 2011.
[4]
S. H. Ferreira, D. C. Bartelt, and L. J. Greene, “Isolation of bradykinin-potentiating peptides from bothrops jararaca venom,” Biochemistry, vol. 9, no. 13, pp. 2583–2593, 1970.
[5]
H. Ni, L. Li, G. Liu, and S. Q. Hu, “Inhibition Mechanism and Model of an Angiotensin I-Converting Enzyme (ACE) Inhibitory hexapeptide from yeast (Saccharomyces cerevisiae),” PLOS ONE, vol. 7, no. 5, Article ID e37077, 2012.
[6]
M. Kuba, C. Tana, S. Tawata, and M. Yasuda, “Production of angiotensin I-converting enzyme inhibitory peptides from soybean protein with Monascus purpureus acid proteinase,” Process Biochemistry, vol. 40, no. 6, pp. 2191–2196, 2005.
[7]
A. J. Mossi, R. L. Cansian, N. Paroul et al., “Morphological characterisation and agronomical parameters of different species of Salvia sp. (Lamiaceae),” Brazilian Journal of Biology, vol. 71, no. 1, pp. 121–129, 2011.
[8]
J. A. Vázquez-Ovando, J. G. Rosado-Rubio, L. A. Chel-Guerrero, and D. A. Betancur-Ancona, “Dry processing of chia (Salvia hispanica L.) flour: chemical and characterization of fiber and protein,” CYTA Journal of Food, vol. 8, no. 2, pp. 117–127, 2010.
[9]
M. R. Segura-Campos, I. M. Salazar-Vega, L. A. Chel-Guerrero, and D. A. Betancur-Ancona, “Biological potential of chia (Salvia hispanica L.) protein hydrolysates and their incorporation into functional foods,” LWT- Food Science and Technology, vol. 50, pp. 723–731, 2013.
[10]
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, pp. 642–646, 2001.
[11]
M. Alaiz, J. L. Navarro, J. Giron, and E. Vioque, “Amino acid analysis of high-performance liquid chromatography after derivatization with diethyl ethoxymethylenemalonate,” Journal of Chromatography, vol. 591, no. 1-2, pp. 181–186, 1992.
[12]
M. J. Cho, N. Unklesbay, F. H. Hsieh, and A. D. Clarke, “Hydrophobicity of bitter peptides from soy protein hydrolysates,” Journal of Agricultural and Food Chemistry, vol. 52, no. 19, pp. 5895–5901, 2004.
[13]
M. Hayakari, Y. Kondo, and H. Izumi, “A rapid and simple spectrophotometric assay of angiotensin-converting enzyme,” Analytical Biochemistry, vol. 84, no. 2, pp. 361–369, 1978.
[14]
D. Montgomery, Dise？o Y Análisis De Experimentos, Limusa-Wiley, Mexico City, Mexico, 2004.
[15]
R. He, H. Ma, W. Zhao et al., “Modeling the QSAR of ACE-Inhibitory peptides with ANN and its applied illustration,” International Journal of Peptides, vol. 2012, Article ID 620609, 9 pages, 2012.
[16]
J. Ruiz-Ruiz, G. Dávila-Ortíz, L. Chel-Guerrero, and D. Betancur-Ancona, “Angiotensin I-converting enzyme inhibitory and antioxidant peptide fractions from hard-to-cook bean enzymatic hydrolysates,” Journal of Food Biochemistry, vol. 37, no. 1, pp. 26–35, 2013.
[17]
J. Pedroche, M. M. Yust, J. Girón-Calle, M. Alaiz, F. Millán, and J. Vioque, “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, vol. 82, no. 9, pp. 960–965, 2002.
[18]
J. Vioque, R. Sánchez-Vioque, A. Clemente, J. Pedroche, J. Bautista, and F. Millan, “Production and characterization of an extensive rapeseed protein hydrolysate,” Journal of the American Oil Chemists' Society, vol. 76, no. 7, pp. 819–823, 1999.
[19]
H. Korhonen, A. Pihlanto-Lepp？l？, P. Rantam？ki, and T. Tupasela, “Impact of processing on bioactive proteins and peptides,” Trends in Food Science and Technology, vol. 9, no. 8-9, pp. 307–319, 1998.
[20]
J. Y. Je, J. Y. Park, W. K. Jung, P. J. Park, and S. K. Kim, “Isolation of angiotensin I converting enzyme (ACE) inhibitor from fermented oyster sauce, Crassostrea gigas,” Food Chemistry, vol. 90, no. 4, pp. 809–814, 2005.
[21]
S. R. Kim and H. G. Byun, “The novel angiotensin I-converting enzyme inhibitory peptides from rainbow trout muscle hydrolysate,” Fisheries and Aquatic Sciences, vol. 15, no. 3, pp. 183–190, 2012.
[22]
X. Y. Mao, J. R. Ni, W. L. Sun, P. P. Hao, and L. Fan, “Value-added utilization of yak milk casein for the production of angiotensin-I-converting enzyme inhibitory peptides,” Food Chemistry, vol. 103, no. 4, pp. 1282–1287, 2007.
[23]
E. G. Tovar-Pérez, I. Guerrero-Legarreta, A. Farrés-González, and J. Soriano-Santos, “Angiotensin I-converting enzyme-inhibitory peptide fractions from albumin 1 and globulin as obtained of amaranth grain,” Food Chemistry, vol. 116, no. 2, pp. 437–444, 2009.
[24]
J. Y. Je, P. J. Park, J. Y. Kwon, and S. K. Kim, “A novel angiotensin I converting enzyme inhibitory peptide from Alaska pollack (Theragra chalcogramma) frame protein hydrolysate,” Journal of Agricultural and Food Chemistry, vol. 52, no. 26, pp. 7842–7845, 2004.
[25]
L. Liu, B. Lu, L. Gong, L. Liu, X. Wu, and Y. Zhang, “Studies on bioactive peptides from Chinese soft-shelled turtle (Pelodiscus sinensis) with functionalities of ACE inhibition and antioxidation,” African Journal of Biotechnology, vol. 11, no. 25, pp. 6723–6729, 2012.
[26]
H. S. Cheung, F. L. Wang, M. A. Ondetti, E. F. Sabo, and D. W. Cushman, “Binding of peptide substrates and inhibitors of angiotensin-converting enzyme. Importance of the COOH-terminal dipeptide sequence,” Journal of Biological Chemistry, vol. 255, no. 2, pp. 401–407, 1980.
[27]
S. Hellberg, L. Eriksson, and J. Jonsson, “Minimum analogue peptide sets (MAPS) for quantitative structure-activity relationships,” International Journal of Peptide and Protein Research, vol. 37, no. 5, pp. 414–424, 1991.
[28]
J. Wu, R. E. Aluko, and S. Nakai, “Structural requirements of angiotensin I-converting enzyme inhibitory peptides: quantitative structure-activity relationship study of Di- and tripeptides,” Journal of Agricultural and Food Chemistry, vol. 54, no. 3, pp. 732–738, 2006.
[29]
A. H. Pripp, T. Isaksson, L. Stepaniak, T. S？rhaug, and Y. Ard？, “Quantitative structure activity relationship modelling of peptides and proteins as a tool in food science,” Trends in Food Science and Technology, vol. 16, no. 11, pp. 484–494, 2005.
[30]
M. A. Ondetti and D. W. Cushman, “Enzymes of the renin-angiotensin system and their inhibitors,” Annual Review of Biochemistry, vol. 51, pp. 283–308, 1982.
[31]
H. G. Byun and S. K. Kim, “Structure and activity of angiotensin I converting enzyme inhibitory peptides derived from alaskan pollack skin,” Journal of Biochemistry and Molecular Biology, vol. 35, no. 2, pp. 239–243, 2002.
[32]
J. S. Hwang and W. C. Ko, “Angiotensin I-converting enzyme inhibitory activity of protein hydrolysates from tuna broth,” Journal of Food and Drug Analysis, vol. 12, no. 3, pp. 232–237, 2004.
[33]
R. Balti, N. Nedjar-Arroume, E. Y. Adjé, D. Guillochon, and M. Nasri, “Analysis of novel angiotensin I-converting enzyme inhibitory peptides from enzymatic hydrolysates of cuttlefish (Sepia officinalis) muscle proteins,” Journal of Agricultural and Food Chemistry, vol. 58, no. 6, pp. 3840–3846, 2010.
[34]
G. N. Kim, H. D. Jang, and C. I. Kim, “Antioxidant capacity of caseinophosphopeptides prepared from sodium caseinate using Alcalase,” Food Chemistry, vol. 104, no. 4, pp. 1359–1365, 2007.
[35]
W. L. Mock, P. C. Green, and K. D. Boyer, “Specificity and pH dependence for acylproline cleavage by prolidase,” Journal of Biological Chemistry, vol. 265, no. 32, pp. 19600–19605, 1990.
[36]
S. Sharma, R. Singh, and S. Rana, “Boactive peptides: a review,” International Journal Bioautomation, vol. 15, no. 4, pp. 223–250, 2011.
