Trypsin Inhibitory Activity of Water Extracts from Ecklonia cava as Affected by Temperature and pH

감태 물 추출물의 Trypsin 저해활성에 대한 열 및 pH 안정성

  • Jung, Seul-A (Dept. of Food Science and Technology / Institute of Food Science, Pukyong National University) ;
  • Kim, Koth-Bong-Woo-Ri (Dept. of Food Science and Technology / Institute of Food Science, Pukyong National University) ;
  • Kim, Min-Ji (Dept. of Food Science and Technology / Institute of Food Science, Pukyong National University) ;
  • Kim, Dong-Hyun (Dept. of Food Science and Technology / Institute of Food Science, Pukyong National University) ;
  • SunWoo, Chan (Dept. of Food Science and Technology / Institute of Food Science, Pukyong National University) ;
  • Kim, Hyun-Jee (Dept. of Food Science and Technology / Institute of Food Science, Pukyong National University) ;
  • Jeong, Da-Hyun (Dept. of Food Science and Technology / Institute of Food Science, Pukyong National University) ;
  • Jeong, Hee-Ye (Dept. of Food Science and Technology / Institute of Food Science, Pukyong National University) ;
  • Kim, Tae-Wan (Dept. of Food Science and Biotechnology, Andong National University) ;
  • Cho, Young-Je (School of Food Science of Biotechnology, Kyungpook National University) ;
  • Ahn, Dong-Hyun (Dept. of Food Science and Technology / Institute of Food Science, Pukyong National University)
  • 정슬아 (부경대학교 식품공학과 / 식품연구소) ;
  • 김꽃봉우리 (부경대학교 식품공학과 / 식품연구소) ;
  • 김민지 (부경대학교 식품공학과 / 식품연구소) ;
  • 김동현 (부경대학교 식품공학과 / 식품연구소) ;
  • 선우찬 (부경대학교 식품공학과 / 식품연구소) ;
  • 김현지 (부경대학교 식품공학과 / 식품연구소) ;
  • 정다현 (부경대학교 식품공학과 / 식품연구소) ;
  • 정희예 (부경대학교 식품공학과 / 식품연구소) ;
  • 김태완 (안동대학교 식품생명공학과) ;
  • 조영제 (경북대학교 식품공학부) ;
  • 안동현 (부경대학교 식품공학과 / 식품연구소)
  • Received : 2012.03.06
  • Accepted : 2012.04.06
  • Published : 2012.06.30


This research was done to verify the inhibitory activity of water extracts from Ecklonia cava (WE-EC) against trypsin and the effects on various temperature and pH conditions. The WE-EC showed high trypsin inhibitory activity of 76, 62 and 60% at concentrations of 5, 2.5 and 1 mg/mL, respectively. In all heat treatments excepted for two conditions, such as $100^{\circ}C$ for 20 min and $121^{\circ}C$ for 15 min, the inhibitory activity was stable compared with the untreated group. With regard to pH stability, the WE-EC showed no significant changes at pH 2~8, but somewhat decreased inhibitory activity was revealed at pH 10. Therefore, the WE-EC could be used in the food industry as a natural trypsin inhibitor.

본 연구는 감태 물 추출물의 일반특성과 trypsin 저해활성을 알아보고 산업적으로 이용가능성을 확인하기 위하여 열 및 pH에 대한 안정성을 실시하였다. 감태 물 추출물의 색도 및 pH를 측정한 결과, 색도는 명도가 86.21로 높고 적색도 및 황색도가 각각 0.38 및 15.49로 낮게 나타났으며 pH는 6.17로 약산성으로 나타났다. 감태 물 추출물의 trypsin 저해활성은 5, 2.5 및 1 mg/mL 농도에서 각각 76.21%, 62.41% 및 60.41%를 나타냈으며 $IC_{50}$값은 0.83 mg/mL이었다. 열 및 pH에 대한 안정성을 측정한 결과, $80^{\circ}C$까지 열처리에 안정하였고 pH 2~8 범위에서 안정하였으나 $100^{\circ}C$$121^{\circ}C$ 열처리와 pH 10에서 활성이 약간 감소하였으나 전체적으로 높은 활성을 유지하여 열 및 pH에 안정한 것을 확인하였다. 이상의 결과를 통해 감태 물 추출물이 지니는 trypsin 저해활성이 열 및 pH에 대해 안정성을 지녀 식품산업에 응용 가능할 것으로 사료된다.



  1. Neurath H. 1984. Evolution of proteolytic enzymes. Science 224: 350-357.
  2. Laskowski MJ, Kato I. 1980. Protein inhibitors of proteinases. Annu Rev Biochem 49: 583-626.
  3. Shewry PR, Lucas JA. 1997. Plant proteins that confer resistance to pests and pathogens. Adv Bot Res 26: 135-192.
  4. Sampaio CA, Oliva ML, Sampaio MU, Batista IFC, Bueno NR, Tanaka AS, Auerswald EA, Fritz H. 1996. Plant proteinase inhibitors. Structure and biochemical applications on plasma kallikrein and related enzymes. Immunopharmacology 32: 62-66.
  5. Walker AJ, Ford L, Majerus MEN, Geoghegan AE, Birch N, Gatehouse JA, Gatehouse AMR. 1997. Characterization proteinase activity of the two-spot ladybird (Adalia bipunctata) and its sensitivity to proteinase inhibitors. Insect Biochem Molec 281: 73-180.
  6. Franco OL, dos Santos RC, Batista JA, Mendes AC, de Araujo MA, Moneratt RG, Grossi-de-Sá MF, de Freitas SM. 2003. Effects of black-eyed pea trypsin/chymotrypsin inhibitor on proteolytic activity and on development of Anthonomus grandis. Phytochemistry 63: 343-349.
  7. Kennedy AR. 1998. Chemopreventive agents: protease inhibitors. Pharmacol Therapeut 78: 167-209.
  8. Oliva MLV, Souza-Pinto JC, Batista IFC, Araujo MS, Silveira VF, Auerswald EA, Mentele R, Eckerskorn C, Sampaio MU, Sampaio CAM. 2000. Leucaena leucocephala serine proteinase inhibitor: primary structure and action on blood coagulation, kinin release and rat paw edema. Biochim Biophys Acta 1477: 64-74.
  9. Lundquist I, Ihse I, Amesjo B. 1976. Carbohydrate metabolism in normal and diabetic rats following long term oral trypsin inhibitor administration. Scand J Gastroentero 11: 369-375.
  10. Min SG, Kim JH, Kim TW, Kim KN. 2003. Isolation and identification of protease producing bacteria in kimchi. Korean J Food Sci Technol 35: 666-670.
  11. Kim SH, Hara S, Hase S, Ikenaka T, Toda H, Kitamura K, Kaizuma N. 1985. Comparative study on amino acid sequences of Kunitz-type soybean trypsin inhibitors, Tia, Tib, and Tic. J Biochem-Tokyo 98: 435-448.
  12. Losso JN. 2008. The biochemical and functional food properties of the Bowman-Birk inhibitor. Crit Rev Food Sci 48: 94-118.
  13. Birk Y. 1985. The Bowman-Birk inhibitor. Trypsin- and chymotrypsin-inhibitor from soybeans. Int J Pept Protein Res 25: 113-131.
  14. Friedman M, Brandon DL. 2001. Nutritional and health benefits of soy proteins. J Agric Food Chem 49: 1069-1086.
  15. Chaudhary NS, Shee C, Islam A, Ahmad F, Yernool D, Kumar P, Sharma AK. 2008. Purification and characterization of a trypsin inhibitor from Putranjiva roxburghii seeds. Phytochemistry 69: 2120-2126.
  16. Klomklao S, Benjakul S, Kishimura H, Osako K, Tanaka M. 2009. A heat-stable trypsin inhibitor in adzuki bean (Vigna angularis): effect of extraction media, purification and biochemical characteristics. Int J Food Sci Tech 45: 163-169.
  17. Klomklao S, Benjakul S, Kishimura H, Chaijan M. 2011. Extraction, purification and properties of trypsin inhibitor from Thai mung bean (Vigna radiata (L.) R. Wilczek). Food Chem 129: 1348-1354.
  18. Kim JM, Park JO, Shin YH. 2008. Purification and characterization of trypsin inhibitor from Alismatis Rhizoma and its binding protein, 10-formyltetrahydrofolate dehydrogenase. Yakhak Hoeji 52: 79-84.
  19. Lee JS, Lee MH, Son HS, Mang YS. 1996. Effects of buckwheat on the activities of pancreatic digestive enzymes in streptozotocin-induced diabetic rats. J Korean Soc Food Sci Nutr 25: 831-838.
  20. Awad NE, Selim MA, Saleh MM, Matloub AA. 2003. Seasonal variation of the lipoidal matters and hypolipidaemic activity of the red alga Corallina officinalis L. Phytother Res 17: 19-25.
  21. Liu JN, Yoshida Y, Wang MQ, Okai Y, Yamachita UB. 1997. B cell stimulating activity of seaweed extracts. Int J Immunopharmaco 19: 135-142.
  22. Kim JY, Kim KH, Suh HS, Choi WC. 1997. Antiinflammatory effects of new chemical compounds, HS-1580 series (HS-1580, HS-1581, HS-1582). J Life Sci 16: 1181-1187.
  23. Ryu BH, Kim DS, Cho K, Sin DB. 1989. Antitumor activity of seaweeds toward Sarcoma-180. Korean J Food Sci Technol 21: 595-600.
  24. Kuda T, Kunii T, Goto H, Suzuki T, Yano T. 2007. Varieties of antioxidant and antibacterial properties of Ecklonia kurome products harvested and processed in the Noto Peninsula, Japan. Food Chem 103: 900-905.
  25. Heo SJ, Park EJ, Lee KW, Jeon YJ. 2005. Antioxidant activities of enzymatic extracts from brown seaweeds. Bioresource Technol 96: 1613-1623.
  26. Jung JY, Kim KBWR, Lee CJ, Kwak JH, Kim MJ, Kim DH, Sunwoo C, Kim TW, Ahn DH. 2011. Inhibitory effect of Ecklonia cava extracts against lipase activity and stability effect of temperature and pH on their activity. J Korean Soc Food Sci Nutr 40: 969-974.
  27. Li Y, Qian ZJ, Ryu BM, Lee SH, Kim MM, Kim SK. 2009. Chemical components and its antioxidant properties in vitro: an edible marine brown alga, Ecklonia cava. Bioorgan Med Chem 17: 1963-1973.
  28. Jung WK, Ahn YW, Lee SH, Choi YH, Kim SW, Choi I, Park SG, Seo SK, Lee SW, Choi IW. 2009. Ecklonia cava ethanolic extracts inhibit lipopolysaccharide-induced cyclooxygenase- 2 and inducible nitric oxide synthase expression in BV2 microglia via the MAP kinase and NF-${\kappa}B$ pathways. Food Chem Toxicol 47: 410-417.
  29. Kong CS, Kim JA, Yoon NY, Kim SK. 2009. Induction of apoptosis by phloroglucinol derivative from Ecklonia Cava in MCF-7 human breast cancer cells. Food Chem Toxicol 47: 1653-1658.
  30. Lee SH, Li Y, Karadeniz F, Kim MM, Kim SK. 2009. $\alpha$-Glucosidase and $\alpha$-amylase inhibitory activities of phloroglucinal derivatives from edible marine brown alga, Ecklonia cava. J Sci Food Agric 89: 1552-1558.
  31. Mabeau, S, Fleurence J. 1993. Seaweed in food products: biochemical and nutritional aspects. Trends Food Sci Tech 4: 103-107.
  32. AOAC. 2000. Official methods of analysis. 17th ed. Association of Official Analytical Chemists, Washington, DC, USA. Vol 3, p 1-25, Vol 31, p 10.
  33. Hummel BCW. 1959. A modified spectrophotometric determination of chymotrypsin, trypsin, and thrombin. Can J Biochem Physiol 37: 1394-1399.
  34. Durre S, Muhammad AR, Shafiq UR, Muhammad AA, Atta UR. 2011. An investigation of phenolic compounds from plant sources as trypsin inhibitors. Nat Prod Res 1: 1-7.
  35. Bitou N, Ninomiya M, Tsujita T, Okuda H. 1999. Screening of lipase inhibitors from marine algae. Lipids 34: 441-445.
  36. Kim SJ, Kweon DH, Lee JH. 2006. Investigation of anti-oxidative activity and stability of ethanol extracts of licorice root (Glycyrrhzia glabra). Korean J Food Sci Technol 38: 584-588.
  37. Ramos VS, Silva GS, Freire MGM, Machado OLT, Parra JRP, Macedo MLR. 2008. Purification and characterization of a trypsin inhibitor from Plathymenia foliolosa seeds. J Agric Food Chem 56: 11348-11355.

Cited by

  1. Isolation of Eckol from Ecklonia cava via Centrifugal Partition Chromatography (CPC) and Characterization of it's Anti-inflammatory Activity vol.48, pp.3, 2015,
  2. Effect of Temperature and pH on Trypsin Inhibitory Activity of Ethanol Extracts from Ecklonia cava vol.27, pp.6, 2012,
  3. Antioxidant and Antiobesity Activity of Natural Color Resources vol.24, pp.6, 2014,
  4. 감태추출물의 기능성원료 표준화를 위한 지표성분 dieckol의 분석법 검증 vol.51, pp.5, 2019,