Effect of phlorotannins isolated from Ecklonia cava on angiotensin I-converting enzyme (ACE) inhibitory activity

  • Wijesinghe, W.A.J.P. ;
  • Ko, Seok-Chun ;
  • Jeon, You-Jin
  • Received : 2010.12.09
  • Accepted : 2011.03.22
  • Published : 2011.04.28


Inhibition of angiotensin I-converting enzyme (ACE) activity is the most common mechanism underlying the lowering of blood pressure. In the present study, five organic extracts of a marine brown seaweed Ecklonia cava were prepared by using ethanol, ethyl acetate, chloroform, hexane, and diethyl ether as solvents, which were then tested for their potential ACE inhibitory activities. Ethanol extract showed the strongest ACE inhibitory activity with an $IC_{50}$ value of 0.96 mg/ml. Five kinds of phlorotannins, phloroglucinol, triphlorethol-A, eckol, dieckol, and eckstolonol, were isolated from ethanol extract of E. cava, which exhibited potential ACE inhibition. Dieckol was the most potent ACE inhibitor and was found to be a non-competitive inhibitor against ACE according to Lineweaver-Burk plots. Dieckol had an inducible effect on the production of NO in EAhy926 cells without having cytotoxic effect. The results of this study indicate that E. cava could be a potential source of phlorotalnnins with ACE inhibitory activity for utilization in production of functional foods.


Angiotensin I-converting enzyme;dieckol;Ecklonia cava;functional foods;phlorotannins


  1. Amenta F, Mignini F, Rabbia F, Tomassoni D, Veglio F. Protective effect of anti-hypertensive treatment on cognitive function in essential hypertension: Analysis of published clinical data. J Neurol Sci 2002;203-204:147-51.
  2. Lee SH, Quian ZJ, Ki SK. A novel angiotensin I converting enzyme inhibitory peptide from tuna frame protein hydrolysate and its antihypertensive effect in spontaneously hypertensive rats. Food Chem 2010;118:96-102.
  3. Jung WK, Mendis E, Je JY, Park PJ, Son BW, Kim HC, Choi YK, Kim SK. Angiotensin I-converting enzyme inhibitory peptide from yellowfin sole (Limanda aspera) frame protein and its antihypertensive effect in spontaneously hypertensive rats. Food Chem 2006;94:26-32.
  4. Bougatef A, Nedjar-Arroume N, Ravallec-Plé R, Leroy Y, Guillochon D, Barkia A, Nasri M. Angiotensin I-converting enzyme (ACE) inhibitory activities of Sardinelle (Sardinella aurita) by-products protein hydrolysates obtained by treatment with microbial and visceral fish serine proteases. Food Chem 2008;111:350-6.
  5. Park EJ, Jhon DY. The antioxidant, angiotensin converting enzyme inhibition activity, and phenolic compounds of bamboo shoot extracts. LWT-Food Sci Technol 2010;4:655-9.
  6. Kim JE, Hwang K, Lee SP. ACE inhibitory and hydrolytic enzyme activities in textured vegetable protein in relation to the solid state fermentation period using Bacillus subtilis HA. Food Sci Biotechnol 2010;19:487-95.
  7. Kim SK, Byun HG, Park PJ, Shahidi F. Angiotensin I converting enzyme inhibitory peptides purified from bovine skin gelatin hydrolysate. J Agric Food Chem 2001;49:2992-7.
  8. Lee JK, Hong S, Jeon JK, Kim SK, Byun HG. Purification and characterization of angiotensin I converting enzyme inhibitory peptides from the rotifer, Brachionus rotundiformis. Bioresour Technol 2009;100:5255-9.
  9. Andrews PR, Carson JM, Caselli A, Spark MJ, Woods R. Conformational analysis and active site modelling of angiotensinconverting enzyme inhibitors. J Med Chem 1985;28:393-9.
  10. Lopez-Fandino R, Otte J, van Camp J. Physiological, chemical and technological aspects of milk-protein-derived peptides with antihypertensive and ACE-inhibitory activity. Int Dairy J 2006;16:1277-93.
  11. Pripp AH, Sorensen R, Stepaniak L, Sørhaug T. Relationship between proteolysis and angiotensin-I-converting enzyme inhibition in different cheeses. LWT-Food Sci Technol 2006;39:677-83.
  12. Miguel M, Alonso MJ, Salaices M, Aleixandre A, Fandino RL, López-Fandiño R. Antihypertensive, ACE-inhibitory and vasodilator properties of an egg white hydrolysate: Effect of a simulated intestinal digestion. Food Chem 2007;104:163-8.
  13. Wu J, Aluko RE, Muir AD. Production of angiotensin I-converting enzyme inhibitory peptides from defatted canola meal. Bioresour Technol 2009;100:5283-7.
  14. Quist EE, Phillips RD, Saalia FK. Angiotensin converting enzyme inhibitory activity of proteolytic digests of peanut (Arachis hypogaea L.) flour. LWT-Food Sci Technol 2009;42:694-9.
  15. Marczak ED, Usui H, Fujita H, Yang Y, Yokoo M, Lipkowski AW, Yoshikawa M. New antihypertensive peptides isolated from rapeseed. Peptides 2003;24:791-8.
  16. Karawita R, Park PJ, Siriwardhana N, Jeon BT, Moon SH, Ahn DK, Cho SK, Jeon YJ. Angiotensin I-converting enzyme (ACE) inhibitory activity of elk (Cervus elaphus) velvet Antler. J Food Sci Nutr 2005;10:239-43.
  17. Nakajima K, Yoshie-Stark Y, Ogushi M. Comparison of ACE inhibitory and DPPH radical scavenging activities of fish muscle hydrolysates. Food Chem 2009;114:884-51.
  18. Athukorala Y, Jeon YJ. Screening for angiotensin 1-converting enzyme inhibitory activity of Ecklonia cava. J Food Sci Nutr 2005;10:134-9.
  19. Spence JT. Challenges related to the composition of functional foods. J Food Compost Anal 2006;19:S4-6.
  20. Cha MH, Lee J, Song MJ. Dieticians' intentions to recommend functional foods: The mediating role consumption frequency of functional foods. Nutr Res Pract 2010;4:75-81.
  21. Li Y, Qian ZJ, Ryu B, Lee SH, Kim MM, Kim SK. Chemical components and its antioxidant properties in vitro: An edible marine brown alga, Ecklonia cava. Bioorg Med Chem 2009;17:1963-73.
  22. Oyamada K, Tsukidate M, Watanabe K, Takahashi T, Isoo T, Terawaki T. A field test of porous carbonated blocks used as artificial reef in seaweed beds of Ecklonia cava. J Appl Phycol 2008;20:863-8.
  23. Shahidi F. Nutraceuticals and functional foods: whole versus processed foods. Trends Food Sci Technol 2009;20:376-87.
  24. Kim MM, Ta QV, Mendis E, Rajapakse N, Jung WK, Byun HG, Jeon YJ, Kim SK. Phlorotannins in Ecklonia cava extract inhibit matrix metalloproteinase activity. Life Sci 2006;9:1436-43.
  25. Lee SH, Han JS, Heo SJ, Hwang JY, Jeon YJ. Protective effects of dieckol isolated from Ecklonia cava against high glucoseinduced oxidative stress in human umbilical vein endothelial cells. Toxicol In Vitro 2010;24:375-81.
  26. Zhang Q, Zhang J, Shen J, Silva A, Dennis DA, Barrow CJ. A simple 96-well microplate method for estimation of total polyphenol content in seaweeds. J Appl Phycol 2006;18:445-50.
  27. Koivikko R, Loponen J, Honkanen T, Jormalainen V. Contents of soluble, cell-wall-bound and exuded phlorotannins in the brown alga Fucus vesiculosus, with implications on their ecological functions. J Chem Ecol 2005;31:195-212.
  28. Kang KA, Lee KH, Park JW, Lee NH, Na HK, Surh YJ, You HJ, Chung MH, Hyun JW. Triphlorethol-A induces heme oxygenase-1 via activation of ERK and NF-E2 related factor 2 transcription factor. FEBS Lett 2007;581:2000-8.
  29. Heo SJ, Ko SC, Cha SH, Kang DH, Park HS, Choi YU, Kim D, Jung WK, Jeon YJ. Effect of phlorotannins isolated from Ecklonia cava on melanogenesis and their protective effect against photo-oxidative stress induced by UV-B radiation. Toxicol In Vitro 2009;23:1123-30.
  30. Chandler SF, Dodds JH. The effect of phosphate, nitrogen, and sucrose on the production of phenolics and solasidine in callus cultures of Solanum laciniatum. Plant Cell Rep 1983;2:205-8.
  31. Cushman DW, Cheung HS. Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung. Biochem Pharmacol 1971;20:1637-48.
  32. Bush K, Henry PR, Slusarchyk DS. Muraceins-Muramyl peptides produced by Nocardia Orientalis as angiotensin-converting enzyme inhibitors. J Antibiot (Tokyo) 1984;37:330-5.
  33. Nath J, Powledge A. Modulation of human neutrophil inflammatory responses by nitric oxide: studies in unprimed and LPS-primed cells. J Leukoc Biol 1997;62:805-16.
  34. Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.
  35. Shibata T, Yamaguchi K, Nagayama K, Kawaguchi S, Nakamura T. Inhibitory activity of brown algal phlorotannins against glycosidases from the viscera of the turban shell Turbo cornutus. Eur J Phycol 2002;37:493-500.
  36. Xu ML, Wang L, Hu JH, Lee SK, Wang MH. Antioxidant activities and related polyphenolic constituents of the methanol extract fractions from Broussonetia papyrifera stem bark and wood. Food Sci Biotechnol 2010;19:677-82.
  37. Handique JG, Baruah JB. Polyphenolic compounds: an overview. React Funct Polym 2002;52:163-88.
  38. Hagerman AE. Extraction of tannin from fresh and preserved leaves. J Chem Ecol 1988;14:453-61.
  39. Porter LJ. Condensed tannins. In: Rowe JW, editor. Natural Products of Woody Plants I. Berlin, Germany: Springer-Verlag; 1989. p.651-90.
  40. Loganayaki N, Siddhuraju P, Manian S. Antioxidant activity of two traditional Indian vegetables: Solanum nigrum L. and Solanum torvum L. Food Sci Biotechnol 2010;19:121-7.
  41. Lee SH, Ko SC, Kang SM, Cha SH, Ahn GN, Um BH, Jeon YJ. Antioxidative effect of Ecklonia cava dried by far infrared radiation drying. Food Sci Biotechnol 2010;19:129-35.
  42. Glombitza KW, Hauperich S. Phlorotannins from the brown alga Cystophora torulosa. Phytochemistry 1997;46:735-40.
  43. Luder UH, Clayton MN. Induction of phlorotannins in the brown macroalga Ecklonia radiata (Laminariales, Phaeophyta) in response to simulated herbivory--the first microscopic study. Planta 2004;218:928-37.
  44. Heo SJ, Jeon YJ. Radical scavenging capacity and cytoprotective effect of enzymatic digests of Ishige okamurae. J Appl Phycol 2008;20:1087-95.
  45. You JS, Sung MJ, Chang KJ. Evaluation of 8-week body weight control program including sea tangle (Laminaria japonica) supplementation in Korean female college students. Nutr Res Pract 2009;3:307-14.
  46. Ahn GN, Kim KN, Cha SH, Song CB, Lee J, Heo MS, Yeo IK, Lee NH, Jee YH, Kim JS, Heu MS, Jeon YJ. Antioxidant activities of phlorotannins purified from Ecklonia cava on free radical scavenging using ESR and $H_2O_2-mediated$ DNA damage. Eur Food Res Technol 2007;226:71-9.
  47. Torruco-Uco J, Chel-Guerrero L, Martinez-Ayala A, Davila-Ortiz G, Betancur-Ancona D. Angiotensin-I converting enzyme inhibitory and antioxidant activities of protein hydrolysates from Phaseolus lunatus and Phaseolus vulgaris seeds. LWT-Food Sci Technol 2009;42:1597-1604.
  48. Appel HM. Phenolics in ecological interactions: the importance of oxidation. J Chem Ecol 1993;19:1521-52.
  49. Kang KA, Lee KH, Chae S, Zhang R, Jung MS, Lee Y, Kim SY, Kim HS, Joo HG, Park JW, Ham YM, Lee NH, Hyun JW. Eckol isolated from Ecklonia cava attenuates oxidative stress induced cell damage in lung fibroblast cells. FEBS Lett 2005;579:6295-304.
  50. Haslam E. Natural polyphenols (Vegetable tannins) as drugs: possible modes of action. J Nat Prod 1996;59:205-15.
  51. Stern JL, Hagerman AE, Steinberg PD, Mason PK. Phlorotanninprotein interactions. J Chem Ecol 1996;22:1877-99.
  52. Shibata T, Yamaguchi K, Nagayama K, Kawagushi S, Nakamura T. Inhibitory activity of brown algal phlorotannins against glycosidases from the viscera of the turban shell Turbo cornutus. Eur J Phycol 2002;37:493-500.
  53. Tsai JS, Lin TC, Chen JL, Pan BS. The inhibitory effects of freshwater clam (Corbicula fluminea, Muller) muscle protein hydrolysates on angiotensin I converting enzyme. Process Biochem 2006;41:2276-81.
  54. Hou YC, Janczuk A, Wang PG. Current trends in the development of nitric oxide donors. Curr Pharm Des 1999;5:417-41.
  55. Abrams J. Beneficial actions of nitrates in cardiovascular disease. Am J Cardiol 1996;77:31C-37C.

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