Angiotensin- I Converting Enzyme Inhibitory Properties of Bovine Casein Hydrolysates in Different Enzymatic hydrolysis Conditions

효소가수분해 조건에 따른 우유 케이신의 Angiotensin-I 전환효소 저해효과

  • Published : 2002.03.01

Abstract

Angiotensiri-I converting enzyme(ACE) catalyst the removal of the C-terminal dipeptide from the angiotensin-I to give the angiotensin-II, a potent peptide that causes constriction of regulation of blood pressure. Recently, ACE inhibitor peptides have been isolated from enzymatic digests of food protein. The aim of this study was to identify bovine casein hydrolysates with ACE inhibitory properties in different enzymatic hydrolysis conditions. The casein were hydrolyzed neutrase, alcalase, protamax, flavourzyme, premed 192, sumizyme MP, sumizyme LP and pescalase alone and with an enzyme combination. Premed 192 produced ACE inhibitory peptides most efficiently. In order to ACE inhibitory peptide produced enzymatic hydrolysis condition were premed 192 added to casein ratio of 1:100(w/w), and incubated at 47$\^{C}$ for 12hrs. Casein hydrolysate gave 50% inhibition(IC$\_$50/ value) of ACE activity at concentration with 248ug/ml(general method) and 265ug/ml(pretreatment method) respectively.

최근 고혈압을 예방하기 위한ACE 저해 펩타이드에 대한 연구는 주로 여러 가지 식품 단백질의 효소 가수분해물로부터 얻어진 펩타이드를 중심으로 이루어지고 있다. 본 연구에서는 케이신을 여러 가지 상업용 단백질분해 효소를 사용하여 ACE저해 효과가 높은 가수분해물 제조시 가수분해 조건이 ACE저해효과에 미치는 영향을 알아보자 하였으며 적정 가수분해 조건을 설정하고자 하였다. ACE 저해효과를 가지는 케이신 가수분해 물을 제조하기 위한 효소 종류, 첨가량 및 가수분해시간은 효소는 Aspergillus oryzae 유래의 promod 192를 사용하고, 효소의 첨가량은 케이신에 대하여 1%, 반응시간은 47$^{\circ}C$에서 12시간으로 하는 것이 적당하였다. 이 때 케이신 가수분해물의 $IC_{50}$/값은 248.71ug/m1(통상법), 265.84ug/ml(전처리법)로서 ACE 저해효과가 높았다.

Keywords

References

  1. Ariyoshi, Y. (1993) Angiotensin- I converting enzyme inhibitors derived from food proteins. Trends in Food Science & Technology, 4, 139-144 https://doi.org/10.1016/0924-2244(93)90033-7
  2. Cheung, H. S., Wang, F. L., Ondetti, M. A., Sabo, E. F. and Cushman, D. W. (1980) Binding of peptide substrates and inhibitors of angiotensin-converting enzyme. The J. of Biol. Chem. 255, 401-407
  3. Cushman, D. W. and Cheung, H. S. (1971) Spectrophotometric assay and properties of the angiotensin converting enzyme of rabbit lung. Biochemical Pharm. 20, 1637-1648 https://doi.org/10.1016/0006-2952(71)90292-9
  4. Hyun, C. K. and Shin, H. K. (2000) Utilization of bovine blood plasma proteins for the production of angiotensin I converting enzyme inhibitory peptides. Process Biochem. 36, 65-71 https://doi.org/10.1016/S0032-9592(00)00176-X
  5. Kinoshita, E., Yamakoshi, J. and Kiluchi, M. (1993) Purification and idientification of angiotensin-I converting enzyme inhibitor from soy sauce. Biosci. Biotech. Biochem. 57, 1107-1081 https://doi.org/10.1271/bbb.57.1107
  6. Kohama, Y., Matsumoto, S., Oka, H., Teramoto, T., Okabe, M. and Mimura, T. (1988) Isolation of angiotensin-converting enzyme inhibitor from tuna muscle. Biochimical and Biophysical Research Communications, 155, 332-337 https://doi.org/10.1016/S0006-291X(88)81089-1
  7. Manjusri, D. and Richard, L. S. (1975) Pulmonary angiotensin converting enzyme. J. Biol. Chem. 250, 6762-6768
  8. Maruyama, S., Mitachi, H., Tanaka, H., Tomizuka, N. and Suzuki, H. (1987) Studies on the active site and antihypertensive activity of angiotensin-I converting enzyme inhibitors derived from casein. Agric. Biol. Chem. 51, 1581-1586
  9. Maruyama, S., Miyoshi, S. and Tanaka, H. (1989) Angiotensin-I converting enzyme inhibitors derived from Ficus carica. Agric. Biol. Chem. 53, 2763-2767
  10. Maruyama, S., Nakagomi, K., Tomizuka, N. and Suzuki, H. (1985) Angiotensin-I converting enzyme inhibitor derived from an enzymatic hydrolysate of casein. II. Isolation and bradykinin-potenting activity on the uterus and the ileum of rats. Agric. Biol. Chem. 49, 1405-1409
  11. Maruyama, S. and Suzuki, H. (1982) A peptide inhibitor of angiotensin converting enzyme in the tryptic hydrolysate of casein. Agric. Biol. Chem. 46, 1393-1394
  12. Matsui, T., Matsufuji, H., Seki, E., Osajima, K., Nakashima, M. and Osajima, Y. (1993) Inhibition of angiotensin I converting enzyme by Bacillus lichenifirmis alkaline protease hydrolysates derived from sardine muscle. Biosci. Biotech. Biochem. 57, 922-925 https://doi.org/10.1271/bbb.57.922
  13. Miyoshi, S., Ishikawa, H., Kaneko, T., Fukui, F., Tanaka, H. and Maruyama, S. (1991) Structures and activity of angiotensin converting enzyme inhibitors in an $\alpha$-zein hydrolysate. Agric. Biol. Chem. 55, 1313-1318
  14. Nakamura, Y., Yamamoto, N., Sakai, K., Okubo, A., Yamazaki, S. and Takano, T. (1995) Purification and characterization of angiotensin- I converting enzyme inhibitor from sour milk. J. of Dairy Sci. 78, 777-783 https://doi.org/10.3168/jds.S0022-0302(95)76689-9
  15. Nakamura, Y., Yamamoto, N., Sakai, K. and Takano, T. (1995) Antihypertensive effect of sour milk and peptide isolated from it that are inhibitors to angiotensin- I converting enzyme. J. of Dairy Sci. 78, 1253-1257 https://doi.org/10.3168/jds.S0022-0302(95)76745-5
  16. SAS (1996) SAS/STAT Software for PC. Releasw 6.11, SAS Institute, Cary, Nc, U.S.A
  17. Webster, J. and Koch, H. F. (1996) Aspects of tolerability of centrally acting antihypertensive drugs. J. of Cardiovascular Pharm. 27, S49-54 https://doi.org/10.1097/00005344-199627003-00007
  18. Yamamoto, N., Akino, A. and Takano, T. (1994) Antihypertensive effect of the peptide derived from casein by on extracellular protenase from Lactobacillus heveticus CP790. J. of Dairy Sci. 77, 917-922 https://doi.org/10.3168/jds.S0022-0302(94)77026-0
  19. Yokoyama, K., Chiba, H. and Yoshikawa, M. (1992) Peptide inhibitors for angiotensin- I converting enzyme from thermolysin digest of dried bonito. Biosci. Biotech. and Biochem. 56, 1541-1545 https://doi.org/10.1271/bbb.56.1541
  20. 上野川修一(1996) 乳の科學. 朝書店, pp. 11
  21. 이은선, 이정희, 박민홍, 허철성, 백영진 (2000) 안지오텐신 전환 효소 효과가 있는 발효유의 제조방법. 대한민국공개특허 2000-0020202
  22. 오세종, 김세헌, 김상교, 백영진, 조경현 (1997) $\kappa$-Casein의 chymosin, Pepsin 및 trypsin 가수분해물에 대한 안지오텐신 변환 효소저해효과 탐색. 한국식품과학회지, 29, 1316-1318
  23. 염동민, 노승배, 이태기, 김선봉, 박영호 (1993) 식품단백질 효소 가수분해물의 Angiotensin I 전환효소 저해작용. 한국영양식량학회지, 22, 226-232
  24. 염동민, 이태기, 변한석, 김선봉, 박영호 (1992) 효소에 의한 고등어 근육단백질 가수분해물의 Angiotensin- I 전환효소 저해작용. 한국수산학회지, 25, 229-235
  25. 原征産, 松本 (1995) 茶の血壓上昇抑制作用. 倉品工業, 38, 78
  26. 川岸瞬朗 (1997) 생체기능조절물질 연구법. 한림원, pp. 121