The Antihypertensive Activity of Angiotensin-Converting Enzyme Inhibitory Peptide Containing in Bovine Lactoferrin


Nai-Yuan Lee;Juei-Tang Cheng;Toshiki Enomoto;Ichiro Nakamura

Key Words

angiotensin-converting enzyme ACE inhibitory peptide ; bovine lactoferrin ; captopril ; spontaneously hypertensive rat


The Chinese Journal of Physiology

Volume or Term/Year and Month of Publication

49卷2期(2006 / 04 / 01)

Page #

67 - 73

Content Language


English Abstract

Angiotensin I-converting enzyme (ACE) inhibitory peptide was isolated from the bovine lactoferrin hydrolysate using peptic hydrolysis by 2-step of reverse-phase high-performance liquid chromatography. This peptide was identified as Leu-Arg-Pro-Val-Ala-Ala and it produced a concentration-dependent inhibition of ACE activity in vitro with an IC50 value of about 4.14 μM. Also, this inhibition was identified as noncompetitive from the Lineweaver-Burk plot. Moreover, the antihypertensive activity of Leu-Arg-Pro-Val-Ala-Ala was investigated by the intravenous injection into spontaneously hypertensive rats (SHRs). A dose-dependent reduction of systolic blood pressure by this peptide was observed at 60 min after injection and it maximally decreased the blood pressure at a rate of 1 nmol/ml/kg. The blood pressure lowering activity of this peptide was calculated as 210% of captopril (10 pmol/ml/kg) that was used as positive control. Otherwise, identification of this peptide in the blood of SHRs was carried out chromatographically. Reduction of blood pressure coincides with the peak peptide concentration in the serum. Thus, we conclude that this peptide inhibits ACE activity in vitro and lowers systolic blood pressure in spontaneously hypertensive rat.

Topic Category 醫藥衛生 > 基礎醫學
  1. Anderson, B.F.,Baker, H.M.,Norris, G.E.,Rice, D.W.,Baker, E.N.(1989).Structure of human lactoferrin: crystallographic structure analysis and refinement at 2.8 A resolution.J. Mol. Biol.,209,711-734.
  2. Ariyoshi, Y.(1993).Angiotensin-converting enzyme inhibitors derived from food proteins.Trends Food Sci. Technol,4,139-143.
  3. Bellamy, W.,Mitsunori, T.,Yamauchi, K.,Wakabayashi, H.,Kawase, K.,Tomita, M.(1992).Identification of the bactericidal domain of lactoferrin.Biochimica. et Biophysica. Acta,1121,130-136.
  4. Bellamy, W.R.,Wakabayashi, H.,Takase, M.,Kawase, K.,Shimamura, S.,Tomita, M.(1993).Role of cell-binding in the antibacterial mechanism of Lactoferricin B..J. Appl. Bacteriol,75,478-484.
  5. Brock, J.H.(2002).The physiology of lactoferrin.Biochem. Cell. Biol.,80,1-6.
  6. Byers, L.D.,Wolfenden, R.(1973).Binding of the by-product analog benzylsuccinic acid by carboxypeptidase A.Biochemistry,12,2070-2078.
  7. Case, D.B.,Atlas, S.A.,Laragh, J.H.,Sealey, J.E.,Sullivan, P.A.,Mckinstry, D.N.(1978).Clinical experience with blockade of the rennin-angiotension-aldosterone system by an oral convertingenzyme inhibitor (captopril) in hypertensive patients.Prog. Cardiovasc. Dis.,21,195-206.
  8. Choi, H.S.,Cho, H.Y.,Yang, H.C.,Ra, K.S.,Suh, H.J.(2001).Angiotensin I-converting enzyme inhibitor from Grifola frondosa.Food Res. Int.,34,177-182.
  9. Cushman, D.W.,Cheung, H.S.(1971).Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung.Biochem. Phamacol,20,1637-1648.
  10. Ehlers, M.R.,Riordan, J.F.(1989).Angiotensin-converting enzyme new concepts converting its biological role.Biochemistry,28,5311-5317.
  11. Fiat, A.M.,Daniele, M.S.,Pierre, J.(1993).Biological active peptides from milk proteins with emphasis on two examples concerning antithrombotic and immunomodulating activities.J. Dairy Sci.,76,301-310.
  12. Gohlke, P.,Linz, W.,Schokens, B.A.,Kuwer, I.,Bartenbach, S.,Chell, A.(1994).Angiotensin converting enzyme inhibition improves cardiac function.Hypertension,26,411-418.
  13. Greene, A.S.,Amaral, S.L.(2002).Microvascular angiogenesis and the renin-angiotensin system.Curr. Hypertens. Rep.,4,56-62.
  14. Kim, S.K.,Bynn, H.G.,Park, P.J.,Shahidi, F.(2001).Angiotensin I converting enzyme inhibitory peptides purified from bovine skin gelatin hydrolysate.J. Agric. Food Chem,49,2992-2997.
  15. Kohama, Y.,Matsumoto, S.,Oka, H.,Teramoto, T.,Okabe, M.,Mimura, T.(1988).Isolation of angiotensin-converting enzyme inhibitor from tuna muscle.Biochem. Biophys. Res. Commun,155,332-337.
  16. Kohmura, M.,Nio, N.,Kubo, K.,Minoshima, Y.,Munekata, E.,Ariyoshi, Y.(1989).Inhibition of angiotensin-converting enzyme by synthetic peptide fragments of human β-casein.Agric. Biol. Chem,53,2107-2114.
  17. Liu, J.C.,Hsu, F.L.,Tsai, J.C.,Chan, P.,Liu, J.Y.H.,Thomas, G.N.,Tomlinson, B.,Lo, M.Y.,Lin, J.Y.(2003).Antihypertensive effects of tannins isolated from traditional Chinese herbs as non-specific inhibitors of angiotensin converting enzyme.Life Sci.,73,1543-1555.
  18. Maruyama, S.,Mitachi, S.,Tanaka, H.,Tomizuka, N.,Suzuki, H.(1987).Studies on the active site antihypertensive activity of angiotensin I-converting enzyme inhibitor derived from casein.Agric. Biol. Chem.,51,1581-1586.
  19. Matoba, N.,Doyama, N.,Yamada, Y.,Maruyama, N.,Utsumi, S.,Yoshikawa, M.(2001).Design and production of genetically modified soybean protein with anti-hypertensive activity by incorporating potent analogue of ovokinin(2-7)..FEBS Lett.,497,50-54.
  20. Matsas, R.,Kenny, A.J.,Turner, A.J.(1984).The metabolism of neuropeptides. The hydrolysis of peptides, including enkephalins, tachykinins and their analogues, by endopeptidase-24.11..Biochem. J.,223,433-440.
  21. Matsumura, N.,Fujii, M.,Takeda, Y.,Shimizu, T.(1993).Angiotensin I-converting enzyme inhibitory peptides derived from bonito bowels autolysate.Biosci. Biotech. Biochem,57,1743-1744.
  22. Miyoshi, S.,Ishikawa, H.,Kaneko, T.,Fukui, F.,Tanaka, H.,Maruyama, S.(1991).Structures and activity of angiotensin-converting enzyme inhibitors in an α-zein hydrolysate.Agric. Biol. Chem,55,1313-1318.
  23. Miyoshi, S.,Kaneko, T.,Yoshizawa, Y.,Fukui, F.,Tanaka, H.,Maruyama, S.(1991).Hypertensive activity of enzymatic α-zein hydrolysate.Agric. Biol. Chem,5,1407-1408.
  24. Nakagomi, K.,Yamada, R.,Ebisu, H.,Sadakane, Y.,Akizawa, T.,Tanimura, T.(2000).Isolation of acein-2, a novel angiotensin Iconverting enzyme inhibitory peptide derived from a tryptic hydrolysate of human plasma.FEBS Lett.,467,235-238.
  25. Nakamura, Y.,Yamamoto, N.,Sakai, K.,Takano, T.(1995).Antihypertensive effect of sour milk and peptides isolated from it that are inhibitors to angiotensin I-converting enzyme.J. Dairy Sci.,78,1253-1257.
  26. Ni, L.,Tao, G.J.,Dai, J.,Wang, Z.,Xu, S.Y.(2001).Separation, purification and identification of angiotensin converting enzyme inhibitory silk fibroin peptide.Se Pu,19,222-225.
  27. Parker, F.,Migliore-Samour, O.,Floch, F.,Zerial, A.,Werner, G.H.,Jolles, J.,Casaretto, M.,Zahn, H.,Jolles, P.(1984).Immunostimulating hexapeptide from human case in amino acid sequence, synthesis and biological properties.Eur. J. Biochem,145,677-682.
  28. Pihlanto-Leppala, A.,Koskinen, P.,Piilola, K.,Tupasela, T.,Korhonen, H.(2000).Angiotensin I-converting enzyme inhibitory properties of whey protein digests: concentration and characterization of active peptides.J. Dairy Res.,67,53-64.
  29. Ramabadran, K.,Bansinath, M.(1989).Pharmacology of β-casomorphins, opioid peptides derived from milk protein.Asia Pac. J. Pharmacol,4,45-49.
  30. Scarborough, R.H.,Rose, J.W.,Hsu, M.H.,Phillips, D.R.,Fried, V.A.,Campbell, A.M.,Manniaai, L.,Charo, I.F.(1991).Barbourin AGpIIb-IIIa specific integrin antagonist from the venom of sistrurus M.Barbouri. J. Biol. Chem.,266,9359-9360.
  31. Sipola, M.,Finckenberg, P.,Santisteban, J.,Korpela, R.,Vapaatalo, H. ,Nurminen, M.L.(2001).Long-term intake of milk peptides attenuates development of hypertension in spontaneously hypertensive rats.J. Physiol. Phatmacol.,52,745-754.
  32. Studdy, P.R.,Lapworth, R.,Bird, R.(1983).Angiotension-converting enzyme and its clinical significance review..J. Clin. Pathol.,36,938-947.
  33. Tauzin, J.,Miclo, L.,Gaillard, J.L.(2002).Angiotensin I-converting enzyme inhibitory peptides from tryptic hydrolysate of bovine α(subscript s2) -casein.FEBS Lett.,6,369-374.
  34. Yokoyama, K.,Chiba, H.,Yoshikawa, M.(1992).Peptide inhibitors for angiotensin I-converting enzyme from thermolysin digest of dried bonito.Biosci. Biotechnol. Biochem.,56,1541-1545.
  35. Yoshii, H.,Tachi, N.,Ohba, R.,Sakamura, O.,Takeyama, H.,Itani, T.(2001).Antihypertensive effect of ACE inhibitory oligopeptides from chicken egg yolks.Comp. Biochem. Physiol. C: Toxicol. Pharmacol.,128,27-33.
  36. Zioudrou, C.,Streaty, R.A.,Klee, W.A.(1979).Opioid peptides derived from food proteins.J. Biol. Chem.,254,2446-2449.