Evaluation of Antioxidant, Antiglycant and ACE-Inhibitory Activity in Enzymatic Hydrolysates of α-Lactalbumin

Full-Text HTML XML Download Download as PDF (Size:1087KB) PP. 84-98
DOI: 10.4236/fns.2017.81006    522 Downloads   690 Views  


Whey proteins are known for their high nutritional value and bioactivity, notably great potential is found on α-lactalbumin. The objective of this study was to evaluate antioxidant, antiglycant and ACE (angiotensin converting enzyme)-inhibitory activity of α-lactalbumin peptides obtained from enzymatic hydrolysis with Alcalase through different hydrolysis conditions. The optimization of enzymatic hydrolysis was studied by response surface methodology variating enzyme:substrate ratio (0.0050%, 0.0525% and 0.1000% w/w) and time (0, 30 and 60 minutes) measuring antioxidant activity by ABTS and ORAC-FL (response variables), founding an augment of this activity with time. Characterization of seven samples showed increasing hydrolysis with time. Hydrolysate obtained with 0.1000% w/w and 60 minutes demonstrated higher antioxidant activity related to greater hydrolysis. This hydrolysate did not show antiglycant activity but promoted advanced glycation end products formation with methylglyoxal and glucose. In contrast, this sample showed 30% of ACE inhibition when compared to Captopril, having great potential by enhancing hydrolysis.

Cite this paper

Fernández-Fernández, A. , López-Pedemonte, T. and Medrano-Fernandez, A. (2017) Evaluation of Antioxidant, Antiglycant and ACE-Inhibitory Activity in Enzymatic Hydrolysates of α-Lactalbumin. Food and Nutrition Sciences, 8, 84-98. doi: 10.4236/fns.2017.81006.


[1] Adjonu, R., Doran, G., Torley, P. and Agboola, S. (2014) Whey Protein Peptides as Components of Nanoemulsions: A Review of Emulsifying and Biological Functionalities. Journal of Food Engineering, 122, 15-27.
[2] Madureira, A.R., Pereira, C.I., Gomes, A.M.P., Pintado, M.E. and Xavier Malcata, F. (2007) Bovine Whey Proteins—Overview on Their Main Biological Properties. Food Research International, 40, 1197-1211.
[3] Adjonu, R., Doran, G., Torley, P. and Agboola, S. (2013) Screening of Whey Protein Isolate Hydrolysates for Their Dual Functionality: Influence of Heat Pre-Treatment and Enzyme Specificity. Food Chemistry, 136, 1435-1443.
[4] Tavares, T.G., Amorim, M., Gomes, D., Pintado, M.E., Pereira, C.D. and Malcata, F.X. (2012) Manufacture of Bioactive Peptide-Rich Concentrates from Whey: Characterization of Pilot Process. Journal of Food Engineering, 110, 547-552.
[5] Demarest, S.J., Boice, J.A, Fairman, R. and Raleigh, D.P. (1999) Defining the Core Structure of the Alpha-Lactalbumin Molten Globule State. Journal of Molecular Biology, 294, 213-221.
[6] Lam, R.S.H. and Nickerson, M.T. (2014) The Effect of pH and Temperature Pre-Treatments on the Physicochemical and Emulsifying Properties of Whey Protein Isolate. LWT—Food Science and Technology, 173, 163-170.
[7] Permyakov, E.A. and Berliner, L.J. (2000) Alpha-Lactalbumin: Structure and Function. FEBS Letters, 473, 269-274.
[8] Gauthier, S.F. and Pouliot, Y. (2003) Functional and Biological Properties of Peptides Obtained by Enzymatic Hydrolysis of Whey Proteins. Journal of Dairy Science, 86, E78-E87.
[9] Korhonen, H. and Pihlanto, A. (2006) Bioactive Peptides: Production and Functionality. International Dairy Journal, 16, 945-960.
[10] Prior, R.L. (2014) Oxygen Radical Absorbance Capacity (ORAC): New Horizons in Relating Dietary Antioxidants/Bioactives and Health Benefits. Journal of Functional Foods, 18, 797-810.
[11] Halliwell, B. and Gutteridge, J.M. (1984) Oxygen Toxicity, Oxygen Radicals, Transition Metals and Disease.The Biochemical Journal, 219, 1-14.
[12] Tironi, V.A. and Anón, M.C. (2010) Amaranth Proteins as a Source of Antioxidant Peptides: Effect of Proteolysis. Food Research International, 43, 315-322.
[13] Contreras, M.D.M., Hernández-Ledesma, B., Amigo, L., Martín-álvarez, P.J. and Recio, I. (2011) Production of Antioxidant Hydrolyzates from a Whey Protein Concentrate with Thermolysin: Optimization by Response Surface Methodology. LWT—Food Science and Technology, 44, 9-15.
[14] Sadat, L., Cakir-Kiefer, C., N’Negue, M.-A., Gaillard, J.-L., Girardet, J.-M. and Miclo, L. (2011) Isolation and Identification of Antioxidative Peptides from Bovine α-Lactalbumin. International Dairy Journal, 21, 214-221.
[15] Hernández-Ledesma, B., Amigo, L., Ramos, M. and Recio, I. (2004) Release of Angiotensin Converting Enzyme-Inhibitory Peptides by Simulated Gastrointestinal Digestion of Infant Formulas .International Dairy Journal, 14, 889-898.
[16] Ahmed, N. (2005) Advanced Glycation Endproducts—Role in Pathology of Diabetic Complications. Diabetes Research and Clinical Practice, 67, 3-21.
[17] Surya, S., Salam, A.D., Tomy, D.V., Carla, B., Kumar, R.A. and Sunil, C. (2014) Diabetes Mellitus and Medicinal Plants—A Review. Asian Pacific Journal of Tropical Disease, 4, 337-347.
[18] Arumugam, G., Manjula, P. and Paari, N. (2013) A Review: Anti Diabetic Medicinal Plants Used for Diabetes Mellitus. Journal of Acute Disease, 2, 196-200.
[19] Ahmed, N and Thornalley, P.J. (2007) Advanced Glycation Endproducts: What Is Their Relevance to Diabetic Complications? Diabetes, Obesity and Metabolism, 9, 233-245.
[20] Szawara-Nowak, D., Koutsidis, G., Wiczkowski, W. and Zielinski, H. (2014) Evaluation of the in Vitro Inhibitory Effects of Buckwheat Enhanced Wheat Bread Extracts on the Formation of Advanced Glycation End-Products (AGEs). LWT—Food Science and Technology, 58, 327-334.
[21] Luevano-Contreras, C. and Chapman-Novakofski, K. (2010) Dietary Advanced Glycation End Products and Aging. Nutrients, 2, 1247-126522.
[22] Peyroux, J. and Sternberg, M. (2006) Advanced Glycation Endproducts (AGEs): Pharmacological Inhibition in Diabetes. Pathologie Biologie, 54, 405-419.
[23] Spotti, M.J., Martinez, M.J., Pilosof, A.M.R., Candioti, M., Rubiolo, A.C. and Carrara, C.R. (2014) Influence of Maillard Conjugation on Structural Characteristics and Rheological Properties of Whey Protein/Dextran Systems. Food Hydrocolloids, 39, 223-230.
[24] Lunceford, N. and Gugliucci, A. (2005) Ilex Paraguariensis Extracts Inhibit AGE Formation More Efficiently than Green Tea. Fitoterapia, 76, 419-427.
[25] Mesías, M., Navarro, M., Martínez-Saez, N., Ullate, M., del Castillo, M.D. and Morales, F.J. (2014) Antiglycative and Carbonyl Trapping Properties of the Water Soluble Fraction of Coffee Silverskin. Food Research International, 62, 1120-1126.
[26] Uribarri, J., Dolores, M., Castillo, D., Pia, M., Maza, D.L., Filip, R., Gugliucci, A., Luevano-Contreras, C., Macias-Cervantes, M.H., Markowicz Bastos, D.H., Medrano, A., Menini, T., Portero-Otin, M., Rojas, A., Sampaio, G.R., Wrobel, K., Wrobel, K. and Garay-Sevilla, M.E. (2015) DietaryAGEs and Their Role in Health and Disease. Advances in Nutrition, 6, 461-473.
[27] Fritz, M., Vecchi, B., Rinaldi, G. and Anón, M.C. (2011) Amaranth Seed Protein Hydrolysates Have in Vivo and in Vitro Antihypertensive Activity. Food Chemistry, 126, 878-884.
[28] Wang, X., Wang, L., Cheng, X., Zhou, J., Tang, X. and Mao, X.Y. (2012) Hypertension-Attenuating Effect of Whey Protein Hydrolysate on Spontaneously Hypertensive Rats. Food Chemistry, 134, 122-126.
[29] Murray, B.A. and FitzGerald, R.J. (2007) Angiotensin Converting Enzyme Inhibitory Peptides Derived from Food Proteins: Biochemistry, Bioactivity and Production. Current Pharmaceutical Design, 13, 773-791.
[30] O’Loughlin, I.B., Murray, B.A., Brodkorb, A., FitzGerald, R.J. and Kelly, P.M. (2014) Production of Whey Protein Isolate Hydrolysate Fractions with Enriched ACE-Inhibitory Activity. International Dairy Journal, 38, 101-103.
[31] Box, G.E.P., Hunter, J.S. and Hunter, W.G. (1978) Statistics for Experimenters. An Introduction to Design, Data Analysis, and Model Building. John Wiley &Sons, New York.
[32] Lowry, O., Rosebrough, N., Farr, A. and Randall, R. (1951) Protein Measurement with the Folin-Phenol Reagent. Journal of Biological Chemistry, 193, 265-275.
[33] Hayakawa, S. and Nakai, S. (1985) Contribution of Hydrophobicity, Net Charge and Sulfhydryl Groups to Thermal Properties of Ovalbumin. Canadian Institute of Food Science and Technology, 18, 290-295.
[34] Kato, A. and Nakai, S. (1980) Hydrophobicity Determined by Fluorescence Probe Method and Its Correlations with Surface Properties of Proteins. Biochemicaet Biophysica Acta, 624, 13-20.
[35] Schagger, H. and von Jagow, G. (1987) Tricine-Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis for the Separation of Proteins in the Range from 1 to 100 kDa. Analytical Biochemistry, 166, 368-379.
[36] Molina Ortiz, S.E. (1997) Modificación enzimática de propiedades funcionales de aislados proteicos de soja. Tesis Doctoral, Universidad Nacional de la Plata, Facultad de Ciencias Exactas, Departamento de Química.
[37] Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M. and Rice-Evans, C. (1999) Antioxidant Activity Applying an Improved ABTS Radical Cation Decolorization Assay. Free Radical Biology & Medicine, 26, 1231-1237.
[38] Ou, B., Hampsch-woodill, M. and Prior, R. (2001) Development and Validation of an Improved Absorbance Capacity Assay Ussing Fluorescein as the Fluorescent. Journal of Agriculture and Food Chemistry, 49, 4619-4626.
[39] Dávalos, A., Bartolomé, B. and Gómez-Cordovés, C. (2005) Antioxidant Properties of Commercial Grape Juices and Vinegars. Food Chemistry, 93, 325-330.
[40] Cushman, D.W. and Cheung, H.S. (1971) Spectrophotometric Assay and Properties of the Angiotensin-Converting Enzyme of Rabbit Lung. Biochemical Pharmacology, 20, 1637-1648.
[41] Kim, Y., Yoon, S., Yu, D., Lonnerdal, B. and Cheung, B. (1999) Novel Angiotensin-I-Converting Enzyme Inhibitory Peptides Derived from Recombinant Human as1-Casein Expressed in Escherichia coli. Journal of Dairy Research, 66, 431-439.
[42] Betty, M., Awuor, O.L., Kirwa, M.E. and Jakim, M.F. (2014) Antioxidative and Functional Properties of Rastrineobolaargentea (Dagaa) Fish Protein Hydrolysate. Discourse Journal of Agriculture and Food Science, 2, 180-189.
[43] Halim, N.R.A., Yusof, H.M. and Sarbon, N.M. (2016) Functional and Bioactive Properties of Fish Protein Hydrolysates and Peptides: A Comprehensive Review. Trends in Food Science and Technology, 51, 24-33.
[44] Jemil, I., Jridi, M., Nasri, R., Ktari, N. Salem, R.B.S., Mehiri, M., et al. (2014) Functional, Antioxidant and Antibacterial Properties of Protein Hydrolysates Prepared from Fish Meat Fermented by Bacillus subtilis A26. Process Biochemistry, 49, 963-972.
[45] Peng, X., Xiong, Y.L. and Kong, B. (2009) Antioxidant Activity of Peptide Fractions from Whey Protein Hydrolysates as Measured by Electron Spin Resonance. Food Chemistry, 113, 196-201.
[46] Saadi, S., Saari, N., Anwar, F., Abdul Hamid, A. and Ghazali, H.M. (2015) Recent Advances in Food Biopeptides: Production, Biological Functionalities and Therapeutic Applications. Biotechnology Advances, 33, 80-116.
[47] Gugliucci, A. (2003) A Practical Method to Study Functional Impairment of Proteins by Glycation and Effects of Inhibitors Using Current Coagulation/Fibrinolysis Reagent Kits. Clinical Biochemistry, 36, 155-158.
[48] Gugliucci, A. (2000) Glicación de proteínas: Rol protagónico de la hiperglicemia en las complicaciones crónicas de la diabetes mellitus. Revista de Medicina de Uruguay, 16, 58-75.

comments powered by Disqus

Copyright © 2017 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.