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Purification and identification of an ACE-inhibitory peptide from walnut protein hydrolysate

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Abstract

Angiotensin I-converting enzyme (ACE) is a dipeptidyl carboxypeptidase. It plays an important physiological role in regulating blood pressure in human bodies. ACE-inhibitory peptides inhibit the activity of ACE, thereby decreasing the tension of blood vessels and the blood volume, thus lowering blood pressure. ACE-inhibitory peptides derived from food proteins due to their safety properties and beneficial effects on human health have attracted more and more attentions on their ACE-inhibitory activity. In the present study, a novel ACE-inhibitory peptide, P-1a1, was homogeneously purified from walnut protein hydrolysate by ultrafiltration, consecutive column chromatography and high performance liquid chromatography. The purified peptide was characterized by Edman degradation, matrix-assisted laser desorption ionization time-of-flight mass spectrophotometer and a liquid-phase peptide sequencer. The amino acid sequence of P-1a1 was determined to be LPGRPPIKPWPL. The potent ACE-inhibitory peptide showed a high ACE-inhibitory activity with the IC50 value of 128.98 μg/mL (95.2 μmol/L). The purified peptide could be used in functional food products as a bioactive component with good ACE-inhibitory activity.

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References

  1. Dorer FE, Kahn JR, Lentz KE, Levine M, Skeggs LT (1974) Hydrolysis of bradykinin by angiotensin-converting enzyme. Circ Res 34:824–827

    Article  CAS  Google Scholar 

  2. He H, Chen X, Sun C, Zhang Y, Zhou B (2006) Analysis of novel angiotensin-I-converting enzyme inhibitory peptides from protease-hydrolyzed marine shrimp Acetes chinensis. J Pept Sci 12:726–733

    Article  CAS  Google Scholar 

  3. Barbosa-Filho JM, Martins VKM, Rabelo LA, Moura MD, Silva MS, Cunha EVL, Souza MFV, Almeida RN, Medeiros IA (2006) Natural products inhibitors of the angiotensin converting enzyme (ACE). A review between 1980 and 2000. Rev Bras Farmacogn 16:421–446

    Article  Google Scholar 

  4. Atkinson AB, Robertson JIS (1979) Captopril in the treatment of clinical hypertension and cardiac failure. Lancet 2:836–839

    Article  CAS  Google Scholar 

  5. Abubakar A, Saito T, Kitazawa H, Kawai Y, Itoch T (1998) Structural analysis of new antihypertensive peptides derived from cheese whey protein by proteinase K digestion. J Dairy Sci 81:3131–3138

    Article  CAS  Google Scholar 

  6. Do JR, Kim KJ, Kim HK (2007) Optimization of enzymatic hydrolysis conditions for production of angiotensin-I converting enzyme inhibitory peptide from casein. Food Sci Biotechnol 16:565–571

    CAS  Google Scholar 

  7. Lo WM, Li-Chan EC (2005) Angiotensin I converting enzyme inhibitory peptides from in vitro pepsin–pancreatin digestion of soy protein. J Agric Food Chem 53:3369–3376

    Article  CAS  Google Scholar 

  8. Suh HJ, Whang JH (1999) A peptide from corn gluten hydrolysate that is inhibitory toward angiotensin I converting enzyme. Biotechnol Lett 21:1055–1058

    Article  CAS  Google Scholar 

  9. Rho SJ, Lee JS, Chung YI, Kim YW, Lee HG (2009) Purification and identification of an angiotensin I-converting enzyme inhibitory peptide from fermented soybean extract. Process Biochem 44:490–493

    Article  CAS  Google Scholar 

  10. Li GH, Le GW, Shi YH, Shrestha S (2004) Angiotensin I-converting enzyme inhibitory peptides derived from food proteins and their physiological and pharmacological effects. Nutr Res 27:469–486

    Article  Google Scholar 

  11. Salcedo CL, Lopez de Mishima BA, Nazareno MA (2010) Walnuts and almonds as model systems of foods constituted by oxidisable, pro-oxidant and antioxidant factors. Food Res Int 43(4):1187–1197

    Article  CAS  Google Scholar 

  12. Aithal BK, Kumar MR, Rao BN, Udupa N, Rao BS (2009) Juglone, a naphthoquinone from walnut, exerts cytotoxic and genotoxic effects against cultured melanoma tumor cells. Cell Biol Int 33(10):1039–1049

    Article  CAS  Google Scholar 

  13. Joseph JA, Shukitt-Hale B, Willis LM (2009) Grape Juice, berries, and walnuts affect brain aging and behavior. J Nutr 139(9):1813S–1817S

    Article  CAS  Google Scholar 

  14. Mukuddem-Petersen J, Stonehouse Oosthuizen W, Jerling JC, Hanekom SM, White Z (2007) Effects of a high walnut and high cashew nut diet on selected markers of the metabolic syndrome: a controlled feeding trial. Br J Nutr 97(6):1144–1153

    Article  CAS  Google Scholar 

  15. Jamdar SN, Rajalakshmi V (2010) Influence of degree of hydrolysis on functional properties, antioxidant activity and ACE inhibitory activity of peanut protein hydrolysate. Food Chem 121:178–184

    Article  CAS  Google Scholar 

  16. Xu H, Hao Y, Qi J, Fan G, Wang K (2009) Study on antioxidant activity and antihypertensive activity of enzymatic hydrolysis products of walnut protein. Innov Ed Farm Prod Process 10:38–42

    Google Scholar 

  17. Cushman DW, Cheung HS (1971) Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung. Biochem Pharmacol 20:1637–1648

    Article  CAS  Google Scholar 

  18. Du M, Wang C, Hu X, Zhao G (2008) Biological properties of different protein extracts from selenium-enriched Ganoderma lucidum. Int J Food Sci Nutr 59(2):134–147

    Article  CAS  Google Scholar 

  19. Du M, Zhao L, Li C, Zhao G, Hu X (2007) Purification and characterization of a novel fungi Se-containing protein from Se-enriched Ganoderma Lucidum mushroom and its Se-dependent radical scavenging activity. Eur Food Res Technol 224:659–665

    Article  CAS  Google Scholar 

  20. Church FC, Swaisgood HE, Porter DH, Catignani GL (1983) Spectrometric assay using O-phthaldialdehyde for determination of proteolysis in milk and isolated milk proteins. J Dairy Sci 66:1219–1227

    Article  CAS  Google Scholar 

  21. Jiang J, Chen S, Ren F, Luo Z, Zeng SS (2007) Yak milk casein as a functional ingredient: preparation and identification of angiotensin-I-converting enzyme inhibitory peptides. J Dairy Res 74:18–25

    Article  CAS  Google Scholar 

  22. Kuba M, Tanaka K, Tawata S, Takeda Y, Yasuda M (2003) Angiotensin I-converting enzyme inhibitory peptides isolated from tofuyo fermented soybean food. Biosci Biotechnol Biochem 67:1278–1283

    Article  CAS  Google Scholar 

  23. Meisel H (1997) Biochemical properties of regulatory peptides derived from milk proteins. Biopolymers 43:119–128

    Article  CAS  Google Scholar 

  24. Megías C, Yust MM, Pedroche J, Lquari H, Girón-Calle J, Aliaiz M, Millán F, Vioque J (2004) Purification of an ACE inhibitory peptide after hydrolysis of sunflower (Helianthus annuus L.) protein isolates. J Agric Food Chem 52:1928–1932

    Article  Google Scholar 

  25. Clare DA, Swaisgood HE (2000) Bioactive milk peptides: a prospectus. J Dairy Sci 83:1187–1195

    Article  CAS  Google Scholar 

  26. 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 59:425–429

    Article  CAS  Google Scholar 

  27. Wu J, Ding X (2001) Hypotensive and physiological effect of angiotensin converting enzyme inhibitory peptides derived from soy protein on spontaneously hypertensive rats. J Agric Food Chem 49:501–506

    Article  CAS  Google Scholar 

  28. 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 54:732–738

    Article  CAS  Google Scholar 

  29. Hernandez-Ledesma B, Isidra R, Mercedes R, Lourdes A (2002) Preparation of ovine and caprine β-lactoglobulin hydrolysates with ACE-inhibitory activity. Identification of active peptides from caprine β-lactoglobulin hydrolysed with thermolysin. Int Dairy J 12:805–812

    Article  CAS  Google Scholar 

  30. Kohmura M (1989) Inhibition of angiotensin-converting enzyme by synthetic peptides of human β-casein. Agric Biol Chem 53:2107–2114

    Article  CAS  Google Scholar 

  31. Pihlanto-Leppala A, Rokka T, Korhone H (1998) Angiotensin I converting enzyme inhibitory peptides derived from bovine milk proteins. Int Dairy J 8:325–331

    Article  Google Scholar 

  32. Matsumura N, Fujii M, Takeda Y, Shimizu T (1993) Isolation and characterization of Angiotensin I-converting enzyme inhibitory peptides derived from bonito bowels. Biosci Biotech Biochem 57(10):1743–1744

    Article  CAS  Google Scholar 

  33. Yamamoto N, Akino A, Takano T (1994) Antihypertensive effect of the peptides derived from casein by an extracellular proteinase form Lactobacillus helveticus CP790. J Dairy Sci 77:917–922

    Article  CAS  Google Scholar 

  34. Gobbetti M, Ferranti P, Smacchi E (2000) Production of angiotensin-I converting enzyme inhibitory peptides in fermented milks started by Lactococcus lactis subsp cremoris FT5. Appl Environ Microbiol 66:3898–3904

    Article  CAS  Google Scholar 

  35. Karaki H, Doi K, Suano S (1990) Antihypertensive effect of tryptic hydrolysate of milk casein in spontaneously hypertensive rats. Comp Biochem Physiol 96(2):367–371

    CAS  Google Scholar 

  36. Mullally MM, Meisel H, Fitzgerald RJ (1997) Identification of novel angiotensin-I-converting enzyme inhibitory peptide corresponding to a tryptic fragment of b-lactoglobulin. FEBS Lett 402:99–101

    Article  CAS  Google Scholar 

  37. 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 83:1434–1440

    Article  CAS  Google Scholar 

  38. Robert MC, Razaname A, Mutter M, Juillerat MA (2004) Identification of angiotensin-I-converting enzyme inhibitory peptides derived from sodium caseinate hydrolysates produced by Lactobacillus helveticus NCC 2765. J Agric Food Chem 52(23):6923–6931

    Article  CAS  Google Scholar 

  39. Quirós A, del Contreras MM, Ramos M, Amigo L, Recio I (2009) Stability to gastrointestinal enzymes and structure–activity relationship of beta-casein-peptides with antihypertensive properties. Peptides 30(10):1848–1853

    Article  Google Scholar 

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Acknowledgments

This work was financially supported by the Fundamental Research Funds for the Central Universities (HIT.BRETIII.201231), the National Natural Science Foundation (31101316 and 31371805), Program of New Century Excellent Talents in University (NCET-11-0796) and the Innovative Talent of Science and Technology Fund in Harbin (2011RFQXN041).

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Compliance with Ethics Requirements

This article does not contain any studies with human or animal subjects.

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Authors and Affiliations

  1. College of Food Science, Northeast Agricultural University, Harbin, 150030, China

    Cong Wang & Lianzhou Jiang

  2. School of Food Science and Engineering, Harbin Institute of Technology, Harbin, 150090, China

    Wei Song & Ming Du

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  1. Cong Wang
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  2. Wei Song
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  3. Lianzhou Jiang
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  4. Ming Du
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Correspondence to Lianzhou Jiang or Ming Du.

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Wang, C., Song, W., Jiang, L. et al. Purification and identification of an ACE-inhibitory peptide from walnut protein hydrolysate. Eur Food Res Technol 239, 333–338 (2014). https://doi.org/10.1007/s00217-014-2227-7

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  • DOI: https://doi.org/10.1007/s00217-014-2227-7

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