Isolation of Angiotensin I-Converting Enzyme Inhibitory Peptide from Chungkookjang

청국장으로부터 Angiotensin I 전환효소 저해 Peptide의 분리

  • Matsui Toshiro (Bioscience and Biotechnology, Graduate School of Agriculture, Kyushu University) ;
  • Yoo Hyung Jae (Department of Life Science, Hoseo University) ;
  • Hwang Jae Sung (Department of Life Science, Hoseo University) ;
  • Lee Dong Seok (Department of Medical Laboratory Science, Inje University) ;
  • Kim Han Bok (Department of Life Science, Hoseo University)
  • 송정이랑 (일본 구주대학원 농학연구원) ;
  • 유형재 (호서대학교 자연과학부 생명과학전공) ;
  • 황재성 (호서대학교 자연과학부 생명과학전공) ;
  • 이동석 (인제대학교 의생명공학대학 임상병리학과) ;
  • 김한복 (호서대학교 자연과학부 생명과학전공)
  • Published : 2004.12.01

Abstract

Chunkookjang, Korean traditional fermented soybean food emerges as a functional food to improve intestinal function and blood circulation. During Chungkookjang fermentation, microorganisms, enzymes, and diverse bioactive compounds increase sharply. Chungkookjang contains diverse oligo-peptides. Formation of peptides was confirmed by SDS-PAGE. Solube fermented soybean in our sample contained Tyr, Gln-Lys, Trp, Gln, and Lys-Pro as major components. Lys-Pro (0.083 mg/100 g sample) was purified by HPLC analysis. Angiotensin I­converting enzyme (ACE) causes hypertension by converting angiotensin I to angiotension II. ACE inhibitory activity of Lys-Pro was determined to be $IC_{50}=32.1\;{\mu}M.$ Whether or not eating Chungkookjang can lower blood pressure was also determined. Sistolic blood pressure dropped by 15 mmHg, and diastolic blood pressure by 8 mmHg 2 hr after a single administration of 20 g of fermented soybean. Chungkookjang might be helpful in improving blood circulation since it has ACE inhibitor and antihypertenisve effect.

Keywords

angiotensin I-converting enzyme;blood pressure;Chungkookjang;peptide;HPLC

References

  1. Kiers, J.L., A.E.A. van Laeken, F.M. Rombouts, and M.J.R. Nout. 2000. In vitro digestibility of Bacillus fermented soya bean. Int. J. Food Microbiol. 60, 163-169
  2. Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685
  3. Rhaleb, N.E., H. Peng, X.P. Yang, Y.H. Liu, D. Mehta, E. Ezan, and O.A. Carretero. 2001. Long-term effect of N-acetyl-serylaspartyl- lysyl-proline on left ventricular collagen deposition in rats with 2-kidney, 1-clip hypertension. Circulation 103, 3136-3141
  4. Urano, T., H. Ihara, K. Umemura, Y. Suzuki, M. Oike, S. Akita. Y. Tsukamoto, I. Suzuki, and A. Takada. 2001. The profibrinolytic enzyme subtilisin NAT purified from Bacillus subtilis cleaves and inactivates plasminogen activator inhibitor type I. J. Biol. Chem. 276, 24690-24696
  5. Matsui, T., K. Tamaya, E. Seki, K. Osajima, K. Matsumoto, and T. Kawasaki. 2002. Val-Tyr as a natural antihypertensive dipeptide can be absorbed into the human circulatory blood system. Clin. Exp. Pharm. Physiol. 29, 204-208
  6. Sarkar, P.K., B. Hasenack, and M.J.R. Nout. 2002. Diversity and functionality of Bacillus and related genera isolated from spontaneously fermented soybeans (Indian Kinema) and locust beans (African Soumbala). Int. J. Food Microbiol. 77, 175-186
  7. 이재중, 이동석, 김한복. 1999. Bacillus licheniformis B1 에 의한 청국장 및 간장발효. 미생물학회지 35, 296-301
  8. Matsui, T., C.H. Li, and Y. Osajima. 1999. Preparation and characterization of novel bioactive peptides responsible for angiotension I-converting enzyme inhibition from wheat germ. J. Peptide Sci. 5, 289-297
  9. Sarkar, F.H. and Y. Li. 2002. Mechanisms of cancer chemoprevention by soy isoflavone genistein. Cancer Metastasis Rev. 21, 265- 280
  10. Chun, H., M. Sasaki, Y. Fujiyama, and T. Bamba. 1996. Effect of peptide chain length on absorption and intact transport of hydrolyzed soybean peptide in rat intestinal everted sac. J. Clin. Biochem. Nutr. 21, 131-140
  11. Sumi, H., H. Hamada, K. Nakanishi, and H. Hiratani. 1990. Enhancement of the fibrinolytic activity in plasma by oral administration of nattokinase. Acta Haematol. 84, 139-143
  12. Suzuki, Y, K. Kondo, Y. Matsumoto, B. Zhao, K. Otsuguro, T. Maeda, Y. Tsukamoto, T. Urano, and K. Umemura. 2003. Dietary supplementation of fermented soybean, natto, suppresses intimal thickening and modulates the lysis of mural thrombi after endothelial injury in rat femoral artery. Life Sci. 73, 1289-1298
  13. Kim, J.S., R.L. Oberle, D.A. Krummel, J.B. Dressman, and D. Fleisher. 1994. Absorption of ACE-inhibitors from small intestine and colon. J. Pharm. Sci. 83, 1350-1356
  14. Shin Z.I., R. Yu, S.A. Park, D.K. Chung, C.W. Ahn, H.S. Nam, K.S. Kim, and H.J. Lee. 2001. His-His-Leu, an angiotensin I converting enzyme inhibitory peptide derived from Korean soybean paste, exerts antihypertensive activity in vivo. J. Agric. Food Chem. 49, 3004-3009
  15. Hirota, A., S. Taki, S. Kawaii, M. Yano, and N. Abe. 2000. 1,1- diphenyl-2-picrylhydrazyl radical-scavenging compounds from soybean miso toward the cancer lines. Biosci. Biotechnol. Biochem. 64, 1038-1040
  16. 이봉기, 장윤수, 이숙이, 정건섭, 최신양 1997. 된장 추출물의 면역조절 기능과 그 작용기전. Kor. J. Immunol. 19, 559-569
  17. 유형재, 이동석, 김한복. 2004. 보리, 쑥, 다시마, 대두 혼합물의 청국장 발효. 미생물학회지 40, 49-53
  18. Kuba. M, K. Tanaka, S. Tawata, Y. Takeda, and M. Yusuda. 2003. Angiotensin I-converting enzyme inhibitory peptides isolated from tofuyo fermented soybean food. Biosci. Biotechnol. Biochem. 67, 1278-1283
  19. Yokoyama, K., H. Chiba, and M. Yoshikawa. 1992. Peptide inhibitors for angiotensin I-converting enzyme from thermolysin digest of dried bonito. Biosci. Biotechnol. Biochem. 56, 1541-1545
  20. Yamamoto, N., A. Akino, and T. Takano. 1994. Antihypertensive effect of the peptides derived from casein by an extracellular proteinase from Lactobacillus helveticus CP790. J. Dairy. Sci. 77, 917-922