Protective Effect of Enzymatic Extracts from Sargassum coreanum on H2O2-induced Cell Damage

  • Ko, Seok-Chun (Department of Marine Life Science, Jeju National University) ;
  • Kang, Sung-Myung (Department of Marine Life Science, Jeju National University) ;
  • Lee, Seung-Hong (Department of Marine Life Science, Jeju National University) ;
  • Ahn, Gin-Nae (Department of Marine Life Science, Jeju National University) ;
  • Kim, Kil-Nam (Jeju Biodiversity Research Institute (JBRI) and Jeju Hi-Tech Industry Development Institute (HiDI)) ;
  • Kim, Yong-Tae (Department of Food Science and Biotechnololgy, Kunsan National University) ;
  • Kim, Jin-Soo (Department of Seafood Science and Technology/Institute of Marine Industry, Gyeongsang National University) ;
  • Heu, Min-Soo (Department of Food Science and Nutrition/Institute of Marine Industry, Gyeongsang National University) ;
  • Jeon, You-Jin (Department of Marine Life Science, Jeju National University)
  • Received : 2010.01.19
  • Accepted : 2010.03.15
  • Published : 2010.03.31


In our previous study, we preliminarily demonstrated that Celluclast and Neutrase extracts exhibited the strongest $H_2O_2$-scavenging activities among five carbohydrases (Viscozyme, Celluclast, Termamyl, Ultraflo and AMG) and five proteases (Kojizyme, Alcalse, Flavourzyme Protamex and Neutrase) extracts. Thus, Celluclast and Neutrase extracts were selected for use in further experiments and were separated into four different molecular weight fractions (<5, 5-10, 10-30 and >30 kDa). Among them, the 5-10 kDa fraction showed the highest $H_2O_2$-scavenging activity. The 5-10 kDa fraction also strongly enhanced cell viability against $H_2O_2$-induced oxidative damage. Furthermore, the fraction reduced the proportion of apoptotic cells induced by $H_2O_2$, as demonstrated by decreased sub-G1 hypodiploid cells and decreased apoptotic body formation by flow cytometry. These results indicated that the 5-10 kDa fraction of the Celluclast and Neutrase extracts from S. coreanum exhibited strong antioxidant activity over $H_2O_2$-mediated cell damage in vitro.


  1. Ahn G, Kim KN, Cha SH, Song CB, Lee JR, Heo MS, Yeo IK, Lee NH, Jee YH, Kim JS, Heu MS and Jeon YJ. 2007. 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 226, 71-79.
  2. Ahn G, Park E, Kim DS, Jeon YJ, Shin T, Park JW, Woo HC, Lee KW and Jee Y. 2008. Anti inflammatory effects of enzymatic extract from Ecklonia cava on TPA-induced ear skin edema. Food Sci Biotechnol 17, 745-750.
  3. Ames B. 1998. Micronutrients prevent cancer and delay aging. Toxicol Lett 102, 5-18.
  4. Athukorala Y, Lee KW, Park EJ, Heo MS, Yeo IK, Lee YD and Jeon YJ. 2003. Reduction of lipid peroxidation and $H_2O_2$-mediated DNA damage by a red alga (Grateloupia filicina) methnolic extract. J Sci Food Agri 85, 251-65.
  5. Athukorala Y and Jeon YJ. 2005. Screening for angiotensin I-converting enzyme inhibitory activity of Ecklonia cava. J Food Sci Nutr 10, 134-139.
  6. Bagchi D, Bagchi M, Stohs SJ, Das DK, Ray SD, Kuszynski CA, Joshi SS and Pruess HG. 2000. Free radicals and grape seed proanthocyanidin extract: importance in human health and disease prevention. Toxicology 148, 187-197.
  7. Chandler SF and Dodds JH. 1983. The effect of phosphate, nitrogen and sucrose on the production of phenolics and solasidine in callus cultures of Solanum laciniatum. Plant Cell Rep 2, 105-108.
  8. Cox DA and Choen ML. 1996. Effects of oxidized low density lipoproteins on vascular contraction and relaxation. Pharmacol Rev 48, 3-9.
  9. El-Habit OHM, Saada HN and Azab KS. 2000. The modifying effect of b-carotene on gamma radiation-induced elevation of oxidative reactions and genotoxicity in male rats. Mutat Res 466, 179-186.
  10. Gonzalez AG, Darias V and Estevez E. 1982. Chemotherapeutic activity of polyhalogenated terpenes from Spanish algae. Planta Med 44, 44-46.
  11. Gschwind M and Huber G. 1995. Apoptotic cell death induced by b-amyloid 1-42 peptide is cell type dependent. J Neurochem 65, 292-300.
  12. Gutteridge JMC. 1995. Lipid peroxidation and antioxidants as biomarker of tissue damage. Clin Chem 41, 1819-1828.
  13. Halliwell B and Aruoma OI. 1991. DNA damage by oxygen-derived species. FEBS Lett 281, 9-19.
  14. Halliwell B, Gutteridge JMC and Cross CE. 1992. Free radicals, antioxidants, and human disease: where are we now. J Lab Clin Med 119, 598-620.
  15. Heo SJ, Ko SC, Kang SM, Kang HS, Kim JP, Kim SH, Lee KW, Cho MG and Jeon YJ. 2008. Cytoprotective effect of fucoxanthin isolated from brown algae Sargassum siliquastrum against $H_2O_2$-induced cell damage. Eur Food Res Technol 228, 145-151.
  16. Heo SJ and Jeon YJ. 2009. Evaluation of diphloretho-hydroxycarmalol isolated from Ishige okamurae for radical scavenging activity and its protective effect against $H_2O_2$-induced cell damage. Process Biochem 44, 412-418.
  17. Heo SJ and Jeon YJ. 2008. Radical scavenging capacity and cytoprotective effect of enzymatic digests of Ishige okamurae. J Appl Phycol 20, 1087-1095.
  18. Heo SJ, Jeon YJ, Lee JH, Kim HT and Lee KW. 2003. Antioxidant effect of enzymatic hydrolyzate from a Kelp, Ecklonia cava. Algae 18, 341-347.
  19. Heo SJ, Park EJ, Lee KW and Jeon YJ. 2005. Antioxidant activities of enzymatic extracts from brown seaweeds. Bioresour Technol 96, 1613-1623.
  20. Kashiwagi M, Mynderse JS, Moore RE and Norton TR. 1980. Antineoplastic evaluation of pacific basin marine algae. J Pharm Sci 69, 735-738.
  21. Kassie F, Parzefall W and Knasmuller S. 2000. Single cell gel electrophoresis assay: a new technique for human biomonitoring studies. Mutat Res 463, 13-31
  22. Kim EK, Lee SJ, Lim BO, Jeon YJ, Song MD, Park TK, Lee KH, Kim B, Lee SR, Moon SH, Jeon BT and Park PJ. 2008. Antioxidative and neuroprotective effects if enzymatic extracts from leaves of Perilla frutescens var. japonica. Food Sci Biotechnol 17, 279-286.
  23. Kim KN, Heo SJ, Song CB, Lee J, Heo MS, Yeo IK, Kang KA, Hyun JW and Jeon YJ. 2006. Protective effect of Ecklonia cava enzymatic extracts on hydrogen peroxide-induced cell damage. Process Biochem 41, 2393-2401.
  24. Ko SC, Kang SM, Ahn G, Yang HP, Kim KN and Jeon YJ. 2009. Antioxidant activity of enzymatic extracts from Sargassum coreanum. J Kor Soc Food Sci Nutr Submitted.
  25. Kosovel V, Avanzini A, Scarcia V and Furlani A. 1988. Algae as possible sources of antitumoral agents. Preliminary evaluation of the in vitro cytostatic activity of crude extracts. Pharmacol Res Commun 20, 27-31.
  26. Kotake-Nara E, Asai A and Nagao A. 2005. Neoxanthin and fucoxanthin induce apoptosis in PC-3 human prostate cancer cells. Cancer Lett 220, 75-84.
  27. Lee SH, Karawita R, Affan A, Lee JB, Lee KW, Lee BJ, Kim DW and Jeon YJ. 2009. Potential of benthic diatoms Achnanthes longipes, Amphora coffeaeformis and Navicula sp. (Bacillariophyceae) as antioxidant sources. Algae 24, 47-55.
  28. Lindenschmidt RC, Trika AF, Guard ME and Witschi HP. 1986. The effect of dietary butylated hydroxyl toluene on liver and colon tumor development in mice. Toxicol 38, 151-160.
  29. Lizard G, Fournel S, Genestier L, Dhedin N, Chaput C, and Flacher M. 1995. Kinetics of plasma membrane and mitochondrial alterations in the cells undergoing apoptosis. Cytometry 21, 275-283.
  30. Mayer AMS and Hmann MT. 2004. Marine pharmacology in 2000: marine compounds with antibacterial, anticoagulant, antifungal, anti-inflammatory, antimalarial, antiplatelet, antiprotozoal, anti-tuberculosis, and antiviral activities; affecting the cardiovascular, immune and nervous systems and other miscellaneous mechanisms of action. Mar Biotechnol 6, 37-52.
  31. Mosmann T. 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Meth 65, 55-63.
  32. Muller HE. 1985. Detection of hydrogen peroxide produced by microorganism on ABTS-peroxide medium. Zentralbl Bakteriol Mikrobiol Hyg 259, 151-158.
  33. Nagayama K, Iwamura Y, Shibata T, Hirayama I and Nakamura T. 2002. Bactericidal activity of phlorotannins from the brown alga Ecklonia kurome. J Antimicrob Chemother 50, 889-893.
  34. Nardella A, Chaubet F, Boisson-vidal C, Blondin C, Durand P and Jozefonvicz J. 1996. Anticoagulant low molecular weight fucans produced by radical process and ion exchange chromatography of high molecular weight fucans extracted from the brown seaweed Ascophyllum nodosum. Carbohydr Res 289, 201-208.
  35. Nicoletti I, Migliorati G, Pagliacci MC, Grignani F and Riccardi C. 1991. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Meth 139, 271-279.
  36. Oki T, Masuda M, Furuta S, Nishibia Y, Terahara N and Suda I. 2002. Involvement of anthocyanins and other phenolic compounds in radical-scavenging activity of purple-fleshed sweet potato cultivars. J Food Sci 67, 1752-1756.
  37. Pena-Ramos EA and Xiong YL. 2002. Antioxidant activity of soy protein hydrolysates in a liposomal system. J Food Sci 67, 2952-2956.
  38. Sherwin ER. 1990. Antioxidants. In Food Additives. Branen AI, Dvidson PM, Salminen S, eds. Marcel Dekker, New York.
  39. Siriwardhana N, Lee KW, Kim SH, Ha JH, Jeon YJ. 2003. Antioxidant activity of Hizika fusifornis on reactive oxygen species scavenging and lipid peroxidation inhibition. Food Sci Technol Int 9, 339-346.
  40. Siriwardhana N, Lee KW, Kim SH, Ha JW, Park GT, Jeon YJ. 2004. Lipid peroxidation inhibitory effects of Hizikia fusiformis methanolic extract on fish oil and linoleic acid. Food Sci Technol Int 10, 65-72.
  41. Wang XW, Zhan Q, Coursen JD, Khan MA, Kontny HU and Yu L. 1999. GADD45 induction of a G2/M cell cycle checkpoint. Proc Natl Acad Sci USA 96, 3706-3711.
  42. Yan X, Chuda Y, Suzuki M and Nagata T. 1999. Fucoxanthin as the major antioxidant in Hizikia fusiformis, a common edible sea weed. Biosci Biotechnol Biochem 63, 605-607.