Abstract
Angiotensin converting enzyme (ACE) and reactive oxygen species are crucial targets for nutritional management of hypertension. The aim of this study is to investigate ACE inhibitory and antioxidative effects of camel and horse milk casein hydrolysates. After casein was isolated from raw camel and horse milk, the purity and molecular weights of the isolated casein were verified using SDS-PAGE prior to digestion with pepsin, trypsin and combined enzymes. The results obtained showed that hydrolysates obtained from pepsin and trypsin combined enzymes exhibited higher ACE-inhibitory activity than individual enzymes. In addition, DPPH radical scavenging activity (%) was significantly (p < 0.05) higher for camel milk casein hydrolysate than horse milk casein hydrolysate. Both hydrolysates displayed significant (p < 0.05) inhibition of lipid peroxidation and hydroxyl radical when compared with standard antioxidant (BHA). Moreover, simulation of casein proteolysis in silico generated fragments with potential ACE inhibitory activity using molecular docking analysis. The present findings suggested that camel and horse milk casein hydrolysates contain bioactive peptides responsible for ACE inhibition and antioxidative effects and thus could be exploited for the treatment and management of hypertension.
Similar content being viewed by others
References
Abdel-Salam AM, Al-Dekheil A, Babkr A, Farahna M, Mousa HM (2013) High fiber probiotic fermented Mare’s milk reduces the toxic effects of mercury in rats. Norteam J Med Sci 2:569–575
Abubakar A, Saito T, Kitazawa H, Kawai Y, Itoh T (1998) Structural analysis of new antihypertensive peptides derived from cheese whey protein by proteinase K digestion. J Dairy Sci 81:3131–3138
Bamdad F, Shin SH, Suh J, Nimalaratne C, Sunwoo H (2017) Anti-inflammatory and antioxidant properties of casein hydrolysates produced using high hydrostatic pressure combined with proteolytic enzymes. Molecules. https://doi.org/10.3390/molecules22040609
Blois MS (1958) Antioxidant determination by the use of a stable free radical. Nature 181:1199–1200
Ceriello A (2008) Possible role of oxidative stress in the pathogenesis of hypertension. Diabetes Care 31(2):S181–S184
Cushman DW, Cheung HS (1971) Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung. Biochem Pharmacol 20:1637–1648
Daliri EB, Deog HO, Byong HL (2017) Bioactive peptides. Foods. https://doi.org/10.3390/foods6050032
Davies DT, Law AJR (1977) An improved method for the quantitative fractionation of casein mixture using ion-exchange chromatography. J Dairy Res 44:213–221
Du Y, Zhao Y, Cai W (2014) Hydroxyl radical scavenging activity of peptide from fish intestine protein by hydrolysis with complex enzyme. Adv J Food Sci Technol 6(1):126–129
Frostegard J, Wu R, Lemne C, Thulin T, Witztum JL, de Faire U (2003) Circulating oxidized low-density lipoprotein is increased in hypertension. Clin Sci 105:615–620
Gasteiger E, Hoogland C, Gattiker A, Duvaud S, Wilkins MR, Appel RD, Bairoch A (2005) Protein identification and analysis tools on the ExPASy server. In: Walker JM (ed) The proteomics protocols handbook. Humana, New York, pp 571–607
Ghasemi S, Khoshgoftarmanesh AH, Hadadzadeh H, Jafari M (2012) Synthesis of Iron-amino acid chelates and evaluation of their efficacy as iron source and growth stimulator for tomato in nutrient solution culture. J Plant Growth Regul 31:498–508
Ghazi L, Drawz P (2017) Advances in understanding the renin-angiotensin-aldosterone system (RAAS) in blood pressure control and recent pivotal trials of RAAS blockage in heart failure and diabetic nephropathy. F1000 Research, 6, 297
Han Z, Zhang W, Luo W, Li J (2016) Novel antioxidant peptides derived from enzymatic hydrolysates of macadamia protein. J Biosci Med 4:6–14
Hernandez-Ledesma B, Garcia-Nebot MJ, Fernandez-Tome S, Amigo L, Recio I (2014) Dairy protein hydrolysates: peptides for health benefits. Int Dairy J 38:82–100
Huma N, Rafiq S, Sameen A, Pasha I, Khan MI (2018) Antioxidant potential of buffalo and cow milk cheddar cheeses to tackle human colon adenocarcinoma (Caco2) cells. Asian-Australas J Anim Sci 31(2):287–292
Jialal I (1998) Evolving lipoprotein risk factors: lipoprotein (a) and oxidized low-density lipoprotein. Clin Chem 44:1827–1832
Kumar R, Chaudhary K, Chauhan JS, Nagpal G, Kumar R, Sharma M, Raghava GPS (2015) An in-silico platform for predicting, screening and designing of antihypertensive peptides. Sci Rep 5:12512
Kumar D, Chatli MK, Singh R, Mehta N, Kumar P (2016) Enzymatic hydrolysis of camel milk casein and its antioxidant properties. Dairy Sci Technol 96:391–404
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lassegue B, Griendling KK (2004) Reactive oxygen species in hypertension: an update. Am J Hypertens 17:852–860
Li Y, Jiang H, Huang G (2017) Protein hydrolysates as promoters of non-haem iron absorption-review. Nutrients 9:609
Lin K, Zhang L, Han X, Xin L, Meng Z, Gong P, Cheng D (2018) Yak milk casein as potential precursor of angiotensin-i-converting enzyme inhibitory peptides based on in silico proteolysis. Food Chem 254:340–347
Mada SB, Reddi S, Kumar N, Kumar R, Kapila S, Kapila R, Trivedi R, Karvande A, Ahmad N (2017a) Antioxidative peptide from milk exhibits anti-osteopenic effects through inhibition of oxidative damage and bone-resorbing cytokines in ovariectomized rats. Nutrition 43–44: 21–31
Mada SB, Reddi S, Kumar N, Kapila S, Kapila R (2017b) Protective effects of casein-derived peptide VLPVPQK against hydrogen peroxide–induced dysfunction and cellular oxidative damage in rat osteoblastic cells. Hum Exp Toxicol 36(4):096032711667829. https://doi.org/10.1177/0960327116678293
Mada SB, Reddi S, Kumar N, Vij R, Yadav R, Kapila S, Kapila R (2018) Casein-derived antioxidative peptide prevents oxidative stress-induced dysfunction in osteoblast cells. PharmaNutrition 6:169–179
Meisinger C, Baumert J, Khuseyinova N, Loewel H, Koenig W (2005) Plasma oxidized low-density lipoprotein, a strong predictor for acute coronary heart disease events in apparently healthy, middle-aged men from the general population. Circulation 112:651–657
Mohanty DP, Mohapatra S, Misra S, Sahu PS (2016) Milk derived bioactive peptides and their impact on human health—a review. Saudi J Biol Sci 23(5):577–583
Muñoz-Durango N, Fuentes CA, Castillo AE, González-Gómez LM, Vecchiola A, Fardella CE, Kalergis AM (2016) Role of the renin-angiotensin-aldosterone system beyond blood pressure regulation: molecular and cellular mechanisms involved in end-organ damage during arterial hypertension. Int J Mol Sci. https://doi.org/10.3390/ijms17070797
Nawaz KAA, David SM, Murugesh E, Thandeeswaran M, Gopikrishnan K, Mahendran R, Palaniswamy M, Angayarkannia J (2017) Identification and in silico characterization of a novel peptide inhibitor of angiotensin converting enzyme from Pigeon pea (Cajanuscajan). Phytomedicine (17) 30130-7
Nisha N, Sreekumar S, Biju C (2016) Identification of lead compounds with cobra venom detoxification activity in andrographis Paniculata (Burm. F.) Nees Through in Silico Method. Int J Pharm Pharm Sci. (8), 212–217
Palaniswamy M, Angayarkanni J, Nandhini B (2012) Angiotensin converting enzyme inhibitory activity and antioxidant properties of goat milk. Hydrolysates Int J Pharm Pharmaceut Sci 4(4):367–370
Paudel KR, Lee UW, Kim DW (2016) Chungtaejeon, a Korean fermented tea, prevents the risk of atherosclerosis in rats fed with a high-fat atherogenic diet. J Integr Med 14(2):134–142
Pihlanto A (2006) Antioxidative peptides derived from milk proteins. Int Dairy J 16:1306–1314
Reddi S, Kumar N, Vij R, Mada SB, Kapila S, Kapila R (2016) Akt drives buffalo casein derived novel peptide mediated osteoblast differentiation. J Nutr Biochem 38:134–144
Ryan MT, Chopra RK (1976) Determination of protein concentration by biuret method. Biochem Biophys Acta 427:337–349
Sachidanandam K, Fagan SC, Ergul A (2005) Oxidative stress and cardiovascular disease: antioxidants and unresolved issues. Cardiovasc Drug Rev 23:115–132
Sarma H, Mattaparthi VSK (2018) Unveiling the transient protein-protein interactions that regulate the activity of human lemur tyrosine kinase-3 (LMTK3) domain by cyclin dependent kinase 5 (CDK5) in breast cancer: an in-silico study. Curr Proteomics 15:62–70
Shanmugam VP, Kapila S, Sonfack TK, Kapila R (2015) Antioxidative peptide derived from enzymatic digestion of buffalo casein. Int Dairy J 42:1–5
Yahya MA, Alhaj OA, Al-Khalifa AS (2017) Antihypertensive effect of fermented skim camel (Camelus Dromedaries) milk on spontaneously hypertensive rats. Nutr Hospistalaria 34(2):416–421
Zidane F, Zeder-Lutz G, Altschuh D, Girardet J, Miclo L, Corbier C, Cakir-Kiefer C (2013) Surface plasmon resonance analysis of the binding mechanism of pharmacological and peptidic inhibitors to human somatic angiotensin I-converting enzyme, biochemistry. http://pubs.acs.org
Zou T, He T, Li H, Tang H, Xia E (2016) The structure-activity relationship of the antioxidant peptides from natural proteins. Molecules 21:72
Acknowledgements
The authors are grateful to the Department of Biochemistry, Ahmadu Bello University Zaria for providing laboratory facilities to carry out this piece of work.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflicts of interest with the contents of this article.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Ugwu, C.P., Abarshi, M.M., Mada, S.B. et al. Camel and Horse Milk Casein Hydrolysates Exhibit Angiotensin Converting Enzyme Inhibitory and Antioxidative Effects In Vitro and In Silico. Int J Pept Res Ther 25, 1595–1604 (2019). https://doi.org/10.1007/s10989-018-09802-2
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10989-018-09802-2