Korean Journal of Fisheries and Aquatic Sciences (한국수산과학회지)

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
권호 표지
DOI QR코드

DOI QR Code

Characteristics of Angiotensin Converting Enzyme Inhibitory Peptides from Thermolysin Hydrolysate of Manila clam, Ruditapes philippinarum Proteins

바지락 단백질 Thermolysin 가수분해물의 Angiotensin Converting Enzyme 저해 Peptide의 특성

  • Lee Tae Gee (Department of Marine Food Industry, Namdo Provincial College of Jeonnam) ;
  • Yeum Dong Min (Department of Food Processing & Baking, Yangsan College) ;
  • Kim Seon Bong (Faculty of Food and Biotechnology, Pukyong National University)
  • 이태기 (남도대학 해양식품산업과) ;
  • 염동민 (양산대학 식품가공제과제빵과) ;
  • 김선봉 (부경대학교 식품생명공학부)
  • Published : 2002.09.01
Download PDF
⟨ Previous Next ⟩

Abstract

The peptides inhibiting angiotensin converting enzyme (ACE) were isolated from the hydrolysate of manila clam (Ruditapes philippinamm) proteins prepared with thermolysin. The thermolysin hydrolysate was pretreated with membrane filter (MW cut-off 10,000) to obtain the peptide fraction with ACE inhibition. The crude peptides were applied to a Sephadex LH-20 column and eluted with $30\%$ methanol. The three active fractions (A, B and C) were collected and concentrated, and then applied to a SP-Toyopearl 650S column equilibrated with distilled water and was eluted with a linear gradient of NaCl concentration (0 to 1 M). The four active fractions (A-1, A-2, B-1 and C-1) were collected and concentrated, and then applied to a SuperQ-Toyopearl 650S column equilibrated with distilled water and was eluted with a linear gradient of NaCl concentration (0 to 1 M). The maximum inhibitory activity was observed in the fraction B-1Q showed the IC_{50} values of 0.748 $\mu$g. The abundant amino acids obtained from active fraction B-1Q were leucine, isoleucine, alanine and threonine.

바지락 단백질의 thermolysin 가수분해물로부터 분자량 한계범위가 10,000 da인 한외여과막을 통과한 저분자량의 물질을 Sephadex LH-20 column을 이용한 겔 크로마토그래피에서 ACE에 대하여 저해 활성을 가지는 3개의 획분을 분취하여 SP-Toyopearl 650S column과 SuperQ-Toyopearl 650S column을 이용한 이온 교환 크로마토그래피에 의하여 4개의 활성 획분을 얻었다. 이 중 가장 높은 절해 활성을 나타내는 획분의 아미노산 조성은 alanine, leucine, isoleucine 및 threonine의 함량이 많았고, 활성 발현에 있어 C 말단 아미노산 잔기로서 중요한 역할을 하는 proline의 함량도 $3.7\%$%인 것으로 나타났다. ACE 저해 활성은 $IC_{50}$ 값이 0.748 $\mu$g이었다.

Keywords

References

  1. Cheung, H.S., F.L. Wang, M.A. Ondetti, E.F. Sabo and D.W. Cushman. 1980. Binding of peptide substrates and inhibitors of angiotensin converting enzyme, Importance of the COOH terminal dipeptide sequence. J. Biol. Chem., 255, 401-407
  2. Cho, YJ., W.S. Cha, S.K. Bok, M.U. Kim, S.S. Chun, U.K. Choi, S.H. Kim and K.S. Park. 2000. Production and separation of angiotensin converting enzyme inhibitor during Natto Fermentation. J. Korean Soc. Food Sci. Nutr., 29, 737-742 (in Korean)
  3. Choi, Y.R., P.J. Park, J.H. Choi, H.G. Byun, I.C. Jeong, S.H. Moonand S.K. Kim. 2000. Purification and characterization of angiotensn I converting enzyme inhibitory peptides from enzymatic hydrolysate of cod liver protein. Korean J. Life Sci, 10, 140-149 (in Korean)
  4. Cushman, D.W., H.S. Cheung, E.F. Sabo and MA Ondetti. 1977. Design of potent competitive inhibitors of angiotensin converting enzyme: carboxyalkanoyl and mercapoalkanoyl amino acids. Biochemistiy, 16, 54-84
  5. Feneira, S.H., D.C. Bartelt and L.J. Greene. 1970. Isolation of bradykinin-potentiating peptides from Bothiops jaraiaca venom. Biochemistiy, 9, 2583-2593 https://doi.org/10.1021/bi00815a005
  6. Frohlich, E.D. 1982. Hemodynamic factors in the pathogenesis and maintenance of hypertension. Fed. Proc. Fed. Am. Soc. Exp. Biol., 41, 2400-2408
  7. Kato, H. and T. Suzuki. 1971. Bradykinin-potentiating peptides from the venom of Agkistmdon halys blomhoffii. Isolation of five bradykinin potentiators and the amino acid sequences of two of them, Potentiators B and C. Biochemistry, 10, 972-980 https://doi.org/10.1021/bi00782a007
  8. Kohama, Y., H. Oka, Y. Kayamori, K. Tsujikawa, T. Mimura, Y. Nagase and M. Satake. 1991. Potent synthetic analogues of angiotensin-converting enzyme inhibitor derived from tuna muscle. Agric. Biol. Chem., 55, 2169-2170 https://doi.org/10.1271/bbb1961.55.2169
  9. Maruyama, S., H. Mitachi, H. Tanaka, N. Tomizuka and H. Suzuki. 1987. Studies on the active site and antihypertensive activity of angiotensin I -converting enzyme inhibitors derived from casein. Agric. Biol. Chem., 51, 1581-1586 https://doi.org/10.1271/bbb1961.51.1581
  10. Matsufuji, H., T. Matsui, E. Seld, K. Osajima, M. Nakashima and Y. Osajima. 1994. Angiotensin I-converting enzyme inhibitory peptides in an alkaline protease hydrolyzate derived from sardine muscle. Biosci. Biotech. Biochem., 58, 2244-2245 https://doi.org/10.1271/bbb.58.2244
  11. Matsumura, N., M. Fujii, Y. Takeda and T. Shimizu. 1993. Isolation and characterization of angiotensin I-converting enzyme inhibitory peptides derived from bonito bowels. Biosci. Biotech. Biochem., 57, 1743-1744 https://doi.org/10.1271/bbb.57.1743
  12. Miyoshi, S., H. Ishikawa, T. Kaneko, F. Fukui, H. Tanaka and S. Maruyama. 1991. Structures and activity of angiotensin converting enzyme inhibitors in an azein hydrolysate. Agric. Biol. Chem., 55, 1313-1318 https://doi.org/10.1271/bbb1961.55.1313
  13. Muramoto, M. and Y. Kawamura. 1991. Antihypertensive peptides derived from rice protein. Shokuhin Kogyo, 11, 18-26 (in Japanese)
  14. Nishikawa, K. 1993. Nonpeptide angiotensin U receptor antagonists. Protein, Nucleic Acid and Enzyme, 38, 239-246 (in Japanese)
  15. Ondetti, M.A, N.J. Williams, E.F. Sabo, J. Pluscec, E.R. Weaver and O. Kocy. 1971. Angiotensin-converting enzyme inhibitors from the venom of Bothmps jararaca. Isolation, elucidation of structure, and synthesis. Biochemistry, 10, 4033-4039 https://doi.org/10.1021/bi00798a004
  16. Osajima, K., S. Sakakibara, T. Sawabe, K. Kitamura, H. Matsuda and Y. Osajima. 1993. Study on separation system of angiotensinconverting enzyme inhibitory peptides originating from sardinemeat. Nippon Shokuhin Kogyo Gakkaishi, 40, 568-576 (in Japanese) https://doi.org/10.3136/nskkk1962.40.568
  17. Oshima, G., H. Shimabukuro and K. Nagasawa. 1979. Peptide inhibitors of angiotensin I -converting enzyme in digests of gelatin by bacterial collagenase. Biochim, Biophys. Acta, 566, 128-137 https://doi.org/10.1016/0005-2744(79)90255-9
  18. Saito, Y., K. Nakamura, A. Kawato and S. Imayasu. 1992. Angiotensin I converting enzyme inhibitors in sake and its by-products. Nippon Nogeikagaku Kaishi, 66, 1081-1087 (in Japanese) https://doi.org/10.1271/nogeikagaku1924.66.1081
  19. Saito, Y., K. Wanezaki (Nakamura), A. Kawato and S. Imayasu. 1994. Antihypertensive effects of peptide in sake and its by-products on spontaneously hypertensive rats. Biosci. Biotech. Biochem., 58, 812-816 https://doi.org/10.1271/bbb.58.812
  20. Saxena, P.R. 1992. Interaction between the renin-angiotensin-aldosterone and sympathetic nervous systems. Joumal of Cardiovascular Pharmacology, 19, S80-S88 https://doi.org/10.1097/00005344-199219006-00013
  21. Sealey, J.E. and J.H. Laragh. 1990. Hypertension: Pathophysiology, diagnosis and management (Edited by Laragh, J.H. and B.M. Brenner). The renin-angiotensin-aldosterone system for normal regulation of blood pressure and sodium and potassium homeostasis, pp. 1287-1317, Raven Press, LTD., New York
  22. Suetsuna, K., M. Yamagami and K. Kuwata. 1988. Inhibitory activity against angiotensin I -converting enzyme of peptides originatingfrom fish and shellfish muscle. Nippon Suisan Gakkaishi, 54, 1853 https://doi.org/10.2331/suisan.54.1853
  23. Suh, H.J., D.B. Suh, S.H. Chung, J.H. Whang, HJ. Sung and H.C. Yang. 1994. Purification of ACE inhibitor from soybean paste. Korean J. Agric. Chem. Biotech., 37, 441-446 (in Korean)
  24. Suh, H.J., S.J. Cho, J.H. Whang, H. Lee and H.C. Yang. 1997. Characterization of angiotensin converting enzyme inhibitorfrom squid hydrolysate. Food and Biotechnology, 6, 122-124
  25. Yeum, D.M., T.G. Lee, H.S. Byun, S.B. Kirn and Y.H. Park. 1992. Angiotensin-1 converting enzyme inhibitory activity of enzymatic hydrolysates of mackerel muscle protein. Bull. Korean Fish. Soc., 25, 229-235 (in Korean)
  26. Yokoyama, K-, H. Chiba and M. Yoshikawa. 1992. Peptide inhibitors for angiotensin-converting enzyme from thermolysin digest of dried bonito. Biosci. Biotech. Biochem., 56, 1541-1545 https://doi.org/10.1271/bbb.56.1541

Cited by

  1. Radical Scavenging Potential of Hydrophilic Phlorotannins of Hizikia fusiformis vol.20, pp.1, 2002, https://doi.org/10.4490/algae.2005.20.1.069
  2. 제주 자생 해양 녹조류와 갈조류 추출물로부터의 항고혈압 활성 vol.35, pp.3, 2002, https://doi.org/10.3746/jkfn.2006.35.3.307
  3. Screening of Extracts from Red Algae in Jeju for Potentials MarineAngiotensin - I Converting Enzyme (ACE) Inhibitory Activity vol.21, pp.3, 2002, https://doi.org/10.4490/algae.2006.21.3.343
  4. The antihypertensive effect and mechanisms of bioactive peptides from Ruditapes philippinarum fermented with Bacillus natto in spontaneously hypertensive rats vol.79, pp.None, 2002, https://doi.org/10.1016/j.jff.2021.104411