Role of Bioactive Peptides in Reducing the Severity of Hypertension with the Inhibition of ACE

  • Muhammad Zohaib Aslam
  • Muhammd Shoaib Aslam
  • Shumaila Firdos
  • Ghulam Ghous
  • Gulistan Firdos
  • Zhao Hongfei
  • Zhang BolinEmail author


Bovine milk protein, fermented with LC (Lactobacillus casei) was used to evaluate its ACE inhibitory activity in-vitro. After evaluating the health status of animals by running hemto-chemical tests, four young dairy animals (22, 26, 30 and 34 months old) were selected on the basis of serum protein level for milk collection. Twenty-two months old animal was considered as control. After co-culturing of skim milk with LC for 24 h, peptide samples were purified via ultrafiltration by using Molecular weight cut-off; 10000 Da (PM-10) membrane. Protein hydrolysis was confirmed by applying ninhydrin reaction and SDS–PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis). Milk of 34-months old animal (D-34) showed greater ACE inhibitory activity in-vitro via ACE inhibitory assay. Moreover, D-34 was further analyzed for its peptides profile by using Q exactive Hybrid Quadrapole-Orbitrap Mass Spectrometry as shown in Graphical abstract. Peptides with higher concentration of Proline, Lysine and aromatic amino acid was identified and higher concentration of hydrophobic aliphatic amino acids in PIN-94 and PIN-332 indicated the usefulness of D-34 as ACE-inhibitor. On the basis of those hydrophobic amino acids it was concluded that Serum protein and milk protein level are interconnected and milk of middle-aged Animals with higher serum protein level is useful to inhibit ACE because with the growth of animal, serum protein contents also increases and those protein contents indicate the health status of the animal. Fermented milk peptides of middle-aged animals can be used as functional food and nutraceutical to reduce hypertension.

Graphical Abstract


Peptides Bovine ACE inhibition Hypertension Fermented milk Hydrophobic amino acids 



This research was done with funding of Beijing Key Laboratory of Food Processing and Safety, School of Biological Sciences and Technology, Beijing Forestry University (China). No funding was received by any agency.

Compliance with Ethical Standards

Conflict of interest

We declare no conflict of interest.


  1. Adler-Nissen J (1979) Determination of the degree of hydrolysis of food protein hydrolysates by trinitrobenzenesulfonic acid. J Agric Food Chem 27:1256–1262Google Scholar
  2. Aengwanich W (2002) Effect of age on hematological values and blood profile of Holstein Friesian crossbred in northeastern Thailand. Suranaree J Sci Technol 9:289–292Google Scholar
  3. Allender PS, Cutler JA, Follmann D, Cappuccio FP, Pryer J, Elliott P (1996) Dietary calcium and blood pressure: a meta-analysis of randomized clinical trials. Ann Intern Med 124:825–831Google Scholar
  4. Al-Saedy KA, Tawfeek FK (2012) Role of age with some physiological and biochemical parameters in local female buffaloes. Iraq J Vet Sci 26:219–223Google Scholar
  5. Appel LJ, Frohlich ED, Hall JE, Pearson TA, Sacco RL, Seals DR, Sacks FM, Smith SC Jr, Vafiadis DK, Van Horn LV (2011) The importance of population-wide sodium reduction as a means to prevent cardiovascular disease and stroke: a call to action from the American Heart Association. Circulation 123:1138–1143Google Scholar
  6. Astawan M, Wahyuni M, Yasuhara T, Yamada K, Tadokoro T, Maekawa A (1995) Effects of angiotensin I-converting enzyme inhibitory substances derived from Indonesian dried-salted fish on blood pressure of rats. Biosci Biotechnol Biochem 59:425–429Google Scholar
  7. Carlene MM, Lawes SVH, Rodgers A (2008) Global burden of blood-pressure-related disease. Lancet 371:1513–1518Google Scholar
  8. Chalé FGH, Ruiz JCR, Fernández JJA, Ancona DAB, Campos MRS (2014) ACE inhibitory, hypotensive and antioxidant peptide fractions from Mucuna pruriens proteins. Process Biochem 49:1691–1698Google Scholar
  9. Cheung H-S, Wang F-l, Ondetti MA, Sabo EF, Cushman DW (1980) Binding of peptide substrates and inhibitors of angiotensin-converting enzyme. Importance of the COOH-terminal dipeptide sequence. J Biol Chem 255:401–407Google Scholar
  10. Cian RE, Alaiz M, Vioque J, Drago SR (2013) Enzyme proteolysis enhanced extraction of ACE inhibitory and antioxidant compounds (peptides and polyphenols) from Porphyra columbina residual cake. J Appl Phycol 25:1197–1206Google Scholar
  11. Cushman D, Cheung H (1971) Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung. Biochem Pharmacol 20:1637–1648Google Scholar
  12. Danaei G, Ding EL, Mozaffarian D, Taylor B, Rehm J, Murray CJ, Ezzati M (2009) The preventable causes of death in the United States: comparative risk assessment of dietary, lifestyle, and metabolic risk factors. PLoS Med 6:e1000058Google Scholar
  13. Doornenbal H, Tong A, Murray N (1988) Reference values of blood parameters in beef cattle of different ages and stages of lactation. Can J Vet Res 52:99Google Scholar
  14. Egan BM, Zhao Y, Axon RN (2010) US trends in prevalence, awareness, treatment, and control of hypertension, 1988–2008. J Am Med Assoc 303:2043–2050Google Scholar
  15. Fagiolo A, Lai O, Alfieri L, Nardon A, Cavallina R (2004) Environmental factors and different managements that influence metabolic, endocrine and immuno responses in water buffalo during lactation. In Proc. Seventh World Buffalo Congress, Manila, pp. 24–26Google Scholar
  16. Gobbetti M, Minervini F, Rizzello CG (2004) Angiotensin i-converting-enzyme-inhibitory and antimicrobial bioactive peptides. Int J Dairy Technol 57:173–188Google Scholar
  17. Gómez-Ruiz J, Ramos M, Recio I (2002) Angiotensin-converting enzyme-inhibitory peptides in Manchego cheeses manufactured with different starter cultures. Int Dairy J 12:697–706Google Scholar
  18. H Z (2004) Study on evaluation index system and method for comprehensive prevention and control community based for hypertension. Huazhong Technology University, WuhanGoogle Scholar
  19. Haque E, Chand R (2008) Antihypertensive and antimicrobial bioactive peptides from milk proteins. Eur Food Res Technol 227:7–15Google Scholar
  20. He R, Ma H, Zhao W, Qu W, Zhao J, Luo L, Zhu W (2012) Modeling the QSAR of ACE-inhibitory peptides with ANN and its applied illustration. Int J Peptides, 2012Google Scholar
  21. Ivanova S (2011) Dynamical changes in the trace element composition of fresh and lyophilized ewe’s milk. Bul J Agric Sci 17:25–30Google Scholar
  22. Ježek J, Nemec M, Starič J, Klinkon M (2011) Age related changes and reference intervals of haematological variables in dairy calves. Bull vet inst pulawy 55:471–478Google Scholar
  23. Jinyu HJZYS (2006) Progress of Antihypertensive Peptide Research [J]. Food Ferment Ind 6:024Google Scholar
  24. Jones ML, Allison RW (2007) Evaluation of the ruminant complete blood cell count. Vet Clin North America 23:377–402Google Scholar
  25. Kawakami A, Kayahara H (1993) Synthesis of leu-lys-tyr derivatives and their interaction with angiotensin I converting enzyme. J Jpn Soc Nutr Food SciGoogle Scholar
  26. Kearney PM, Whelton M, Reynolds K, Muntner P, Whelton PK, He J (2005) Global burden of hypertension: analysis of worldwide data. The Lancet 365:217–223Google Scholar
  27. Kim JH, Jeong LDH, Chung SC K.S. & Lee JS (2004) Characterization of antihypertensive angiotensin I-converting enzyme inhibitor from Saccharomyces cerevisiae. J Microbiol Biotechnol 16:1318–1323Google Scholar
  28. Klinkon M, Simčič M, Ježek J, Klinkon Z, Kompan D (2014) Factors affecting blood parameters of autochthonous Cika cattle. Acta Agraria Kaposváriensis 14:141–145Google Scholar
  29. Korhonen H, Pihlanto A (2006) Bioactive peptides: production and functionality. Int Dairy J 16:945–960Google Scholar
  30. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680Google Scholar
  31. Mahmoud MI (1994) Physicochemical and functional properties of protein hydrolysates in nutritional products. Food Technol 48:89–113Google Scholar
  32. Matthews D, Payne J (1980) Transmembrane transport of small peptides. In Current topics in membranes and transport, Vol. 14 Elsevier, New York, pp. 331–425Google Scholar
  33. Meisel H (1998) Overview on milk protein-derived peptides. Int Dairy J 8:363–373Google Scholar
  34. Mitruka BM, Rawnsley HM (1977) Clinical biochemical and hematological reference values in normal experimental animals. Clin Biochem Hematol Ref Values Norm Exp AnimGoogle Scholar
  35. Mohri M, Sharifi K, Eidi S (2007) Hematology and serum biochemistry of Holstein dairy calves: age related changes and comparison with blood composition in adults. Res Vet Sci 83:30–39Google Scholar
  36. Murray B, FitzGerald R (2007) Angiotensin converting enzyme inhibitory peptides derived from food proteins: biochemistry, bioactivity and production. Curr Pharm Des 13:773–791Google Scholar
  37. Omer R, Eltinay A (2008) Microbial quality of camel’s raw milk in central & southern regions of United Arab Emirates. Emirates J Food Agric, 76–83Google Scholar
  38. Perkovic V, Huxley R, Wu Y, Prabhakaran D, MacMahon S (2007) The burden of blood pressure-related disease: a neglected priority for global health. Hypertension 50:991–997Google Scholar
  39. Roberts PR, Burney J, Black KW, Zaloga GP (1999) Effect of chain length on absorption of biologically active peptides from the gastrointestinal tract. Digestion 60:332–337Google Scholar
  40. Rodríguez-Figueroa J, González-Córdova A, Torres-Llanez M, Garcia H, Vallejo-Cordoba B (2012) Novel angiotensin I-converting enzyme inhibitory peptides produced in fermented milk by specific wild Lactococcus lactis strains. J Dairy Sci 95:5536–5543Google Scholar
  41. Saito T, Nakamura T, Kitazawa H, Kawai Y, Itoh T (2000) Isolation and structural analysis of antihypertensive peptides that exist naturally in Gouda cheese. J Dairy Sci 83:1434–1440Google Scholar
  42. Schanbacher F, Talhouk R, Murray F, Gherman L, Willett L (1998) Milk-borne bioactive peptides. Int Dairy J 8:393–403Google Scholar
  43. Sparrenberger F, Cichelero F, Ascoli A, Fonseca F, Weiss G, Berwanger O, Fuchs S, Moreira L, Fuchs F (2009) Does psychosocial stress cause hypertension? A systematic review of observational studies. J Hum Hypertens 23:12Google Scholar
  44. Suetsuna K, Nakano T (2000) Identification of an antihypertensive peptide from peptic digest of wakame (Undaria pinnatifida). J Nutr Biochem 11:450–454Google Scholar
  45. Thaker D, Frech F, Suh D-C, Aranda J, Shin H, Rocha R (2005) P-308: Prevalence of hypertension and ethnic differences in sociodemographic and cardiovascular health characteristics of US hypertensives. Am J Hypertens 18:117A–117AGoogle Scholar
  46. Udenigwe CC, Aluko RE (2011) Chemometric analysis of the amino acid requirements of antioxidant food protein hydrolysates. Int J Mol Sci 12:3148–3161Google Scholar
  47. Vermeirssen V, Van Camp J, Verstraete W (2004) Bioavailability of angiotensin I converting enzyme inhibitory peptides. Br J Nutr 92:357–366Google Scholar
  48. Wu X, Duan X, Gu D, Hao J, Tao S, Fan D (1995) Prevalence of hypertension and its trends in Chinese populations. Int J Cardiol 52:39–44Google Scholar
  49. Wu J, Aluko RE, Nakai S (2006) Structural requirements of angiotensin I-converting enzyme inhibitory peptides: quantitative structure – activity relationship study of di-and tripeptides. J Agric Food Chem 54:732–738Google Scholar
  50. Wu Y, Huxley R, Li L, Anna V, Xie G, Yao C, Woodward M, Li X, Chalmers J, Gao R, Kong L, Yang X, China NSC, China NWG (2008) Prevalence, awareness, treatment, and control of hypertension in China: data from the China National Nutrition and Health Survey 2002. Circulation 118:2679–2686Google Scholar
  51. Yamamoto N, Maeno M, Takano T (1999) Purification and characterization of an antihypertensive peptide from a yogurt-like product fermented by Lactobacillus helveticus CPN4. J Dairy Sci 82:1388–1393Google Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Muhammad Zohaib Aslam
    • 1
  • Muhammd Shoaib Aslam
    • 2
  • Shumaila Firdos
    • 3
  • Ghulam Ghous
    • 4
  • Gulistan Firdos
    • 4
  • Zhao Hongfei
    • 1
  • Zhang Bolin
    • 1
    Email author
  1. 1.Beijing Key Laboratory of Food Processing and SafetyBeijing Forestry UniversityBeijingChina
  2. 2.Department of Forest economics and managementBeijing Forestry UniversityBeijingChina
  3. 3.Punjab Livestock and dairy development departmentLahorePakistan
  4. 4.Institute of Chemical SciencesBahauddin Zakariya UniversityMultanPakistan

Personalised recommendations