組織蛋白質酶水解物之抗高血壓胜肽的分離與特性分析

Purification and Characterization of an Angiotensin I-Converting Enzyme Inhibitory Peptide from Catheptic Hydrolysates

10.29770/JTCMT.200909.0007

李嘉展(Jai-Jaan Lee)；聶方珮(Fang-Pei Nieh)；孫明輝(Ming-Hui Sun)；彭清勇(Ching-Yung Pong)

組織蛋白酶 ； 竹筴魚 ； 機能性食品 ； cathepsin ； scad ； functional food

台北海洋技術學院學報

2卷2期（2009 / 09 / 01）

149 - 160

繁體中文

本研究以竹筴魚肌肉爲材料，純化出內生性組織蛋白酶B，其純化倍率、產率和分子量分別爲7，814倍、3.8%和27,500Da，利用純化的蛋白酶B水解肌動凝蛋白質，其水解產物經Shephadex G-25膠過濾層析分劃出四個分劃物，其分子量分別爲1,700、1,400、900和550Da，其血管升壓素轉換酶半抑制濃度分別爲592.6、190.3、91.6和65.8μg peptide/ml，本研究結果顯示竹筴魚肌肉蛋白質經組織蛋白酶B水解後的產物具有血管升壓素轉換酶抑制活性，未來將可應用於降高血壓機能性食品的開發。

The study was to purify cathepsin B from horse mackerel (Trachurus japonicus) and fractionate the angiotensin I-converting enzyme (ACE) inhibitory peptides from catheptic hydrolysates. The purification fold, yield and molecular weight of horse mackerel cathepsin B were 7,814 fold, 3.8% and 27,500Da respectively. Four fractions with ACE-inhibitory activity appeared on Shephadex G-25 chromatography. Their molecular weight were 1,700, 1,400, 900and 550Da respectively. The IC50 of four hydrolysates were 592.6, 190.3, 91.6 and 65.8μg peptide/ml. The results suggest that horse mackerel protein hydrolysates by cathepsin B might be a potential ingredient for the formulation of hypotensive functional foods.

1. Arihara, K.,Nakashima, Y.,Mukai, T.,Ishikawa, S.,Itoh, M.(2001).Peptide inhibirors for angiotensin I-converting enzyme from enzymatic hydrolysates of porcine skeletal muscle protieins.Meat Sci.,57,319-324.
2. Barrett, A. J.,Kirschke, H.(1981).Cathepsin B, cathepsin H and cathepsin L.Methods Enzymol.,80,535-561.
3. Bougatef, A.,Nedjar-Arroume, N.,Ravallec-Ple, R.,Leroy, Y.,Guillochon, D.,Barkia, A.,Nasri, M.(2008).Angiotensin I-converting enzyme (ACE) inhibitory activities of sardinelle (Sardinella aurita) by-products protein hydrolysates obtained by treatment with microbial and visceral fish serine proteases.Food Chem.,111,350-356.
4. Byun, H. G.,Kim, S. K.(2001).Purification and characterization of angiotensin I converting enzyme (ACE) inhibitory peptides from Alaska pollack (Theragra chalcogramma) skin.Process Biochem.,36,1155-1162.
5. Chen, J. R.,Yang, S. C.,Suetsana, K.,Chao, J. C. J.(2004).Soybean protein-derived hydrolysate affects blood pressure in spontaneously hypertensive rats.J. Food Biochem.,28,61-73.
6. Cheung, H. S.,Chushman, D. W.(1971).Spectrophotometric assay and properties of the angiotensin converting enzyme of rabbit lung.Biochem. Pharmacol.,20,1637-1648.
7. Clare, D. A.,Swaisgood, H. E.(2000).Bioactive milk peptides: a prospectus.J. Dairy Sci.,83,1187-1195.
8. Fujita, H.,Yasumoto, R.,Hasegawa, M.,Ohshima, K.(2001).Effects of an ace-inhibitory agent, katsuobushi oligopeptide, in the spontaneously hypertensive rat and in borderline and mildly hypertensive subjects.Nutr. Res.,21,1149-1158.
9. Fujita, H.,Yoshikawa, M.(1999).LKPNM: a prodrug type ACE-inhibitory peptide derived from fish protein.Immunopharmacology,44,123-127.
10. Fujita, H.m,Yokoyama, K.,Yoshikawa, M.(2000).Classification and antihypertensive activity of angiotensin I-converting enzyme inhibitory peptides derived from food proteins.J. Food Sci.,65,564-569.
11. He, H. L.,Chen, X. L.,Wu, H.,Sun, C. Y.,Zhang, Y. Z.,Zhou, B. C.(2007).High throughput and rapid screening of marine protein hydrolysates enriched in peptides with angiotensin-I-converting enzyme inhibitory activity by capillary electrophoresis.Bioresource Technol.,98,3499-3505.
12. Hyun, C. K.,Shin, H. K.(2000).Utilization of bovine plasma proteins for the production of angiotensin I converting enzyme inhibitory peptides.Process Biochem.,36,65-71.
13. Israaili, Z. H.,Hall, W. D.(1992).Cough and angioneurotic edema associated with angiotensin converting enzyme inhibitor therapy: A review of the literature and pathophysiology.Ann. Intern. Med.,117,234-242.
14. Je, J. Y.,Park, P. J.,Kwon, J. Y.,Kim, S. K.(2004).A novel angiotensin I-converting enzymeinhibitory peptide from Alaska Pollack (Theragra chalcogramma) frame protein hydrolysate.J. Agri. Food Chem.,52,7842-7825.
15. Jiang, S. H.,Lee, Jai-Jaan,Chen, H. C.(1996).Proteolysis of actomyosin by cathepsins B, L, L-like, and X from mackerel (Scomber australasicus).J. Agri. Food Chem.,44(3),769-773.
16. Jiang, S. T.,Lee, Jai-Jaan,Chen, H. C.(1994).Purification and characterization of cathepsin B from ordinary muscle of mackerel (Scomber australasicus).J. Agri. Food Chem.,42(5),1073-1079.
17. Kapel, R.,Rahhou, E.,Lecouturier, D.,Guillochon, D.,Dhuslster, P.(2006).Characterization of an antihypertensive peptide from an alfalfa white protein hydrolysate produced by a continuous enzymatic membrane reactor.Process Bioch.,41,1961-1966.
18. Kim, S. K.,Byun, H. G.,Park, P. J.,Shahidi, F.(2001).Angiotensin I converting enzyme inhibitory peptides purified from bovine skin gelatin hydrolysate.J. Agri. Food Chem.,49,2992-2997.
19. Kohama, Y.,Matsumoto, S.,Oka, H.,Teramoto, T.,Okabe, M.,Mimura, Y.(1988).Isolation of angiotensin-converting enzyme-inhibitor from tuna muscle.Biochem. Biophys. Res. Commun.,155,332-337.
20. Kohama, Y.,Oka, H,Kayamori, Y.,Tsujikawa, K.,Mimura, T.,Nagase, Y.,Satake, M.(1991).Poten synthetic analogs of angiotensin-converting enzyme-inhibitor derived from tuna muscle.Agric. Biol. Chem.,55,2169-2170.
21. Li, C. H.,Matsui, T.,Matsumoto, K.,Yamasaki, R.,Kawasaki, T.(2002).Latent production of angiotensin I-converting enzyme inhibitors from buckwheat protein.J. Pept. Sci.,8,267-274.
22. Li, G. H.,Le, G. W.,Shi, Y. H.,Shrestha, S.(2004).Angiotensin I-converting enzyme inhibitory peptides derived from food proteins and their physiological and pharmacological effects.Nutr. Res.,24,469-486.
23. Li, G. H.,Wan, J. Z.,Le, G. W.,Shi, Y. H.(2006).Novel angiotensin I-converting enzyme inhibitory peptides isolated from alcase hydrolysate of mung bean protein.J. Pept. Sci.,12,509-514.
24. Maruyama, S.,Miyoshi, S.,Kaneko, T.,Tanaka, H.(1989).Angiotensin-I-converting enzyme inhibitory activities of synthetic peptides related ot the tandem repeated sequence of a miaze indosperm protein.Agric. Biol. Chem.,53,1077-1081.
25. Matsui, T.,Li, C. H.,Osajima, Y.(1999).Preparation and characterization of novel bioactive peptides responsible for angiotensinI-converting enzyme inhibition from wheat germ.J. Pept. Sci.,5,289-297.
26. Meisel, H.(1997).Biochemical properties of bioactive peptides derived from milk proteins: Potential nutraceuticals for food and pharmaceutical applications.Livest. Prod. Sci.,50,125-138.
27. Miyoshi, S.,Ishikawa, H.,Kaneko, T.,Fukui, F.,Tanaka, H.,Maruyama, S.(1991).Structures and activity of angiotensin-converting enzyme inhibitors in an alpha-zein hydrolysate.Agric. Biol. Chem.,55,1313-1318.
28. Nakano, D.,Ogura, K.,Miyakoshi, M.,Ishii, F.,Kawanishi, H.,Kurumazuka, D.,Kwak, C.,Ikemura, K.,Takaoka, M.,Moriguchi, S.,Iino, T.,Kusumoto, A.,Asami, S.,Shibata, H.,Kiso, Y.,Matsumura, Y.(2006).Antihypertensive effect of angiotensin I-converting enzyme inhibitory peptides from a sesame protein hydrolysate in spontaneously hypertensive rats.Biosci. Biotechnol. Biochem.,70,1118-1126.
29. Noguchi, S.(1974).Tokyo, Japan,Sophia University.
30. Ondetti, M. A.,Cushman, D. W.(1982).Enzymes of the rennin-angiotensin system and their inhibitors.Annu. Rev. Bioch.,51,283-308.
31. Ono, S.,Hosokawa, M.,Miyashita, K.,Takahashi, K.(2003).Isolation of peptides with angiotensin I-converting enzyme inhibitory effect derived from hydrolysate of upstream chem. Salmon muscle.J. Food Sci.,68,1611-1614.
32. Ono, S.,Hosokawa, M.,Miyashita, K.,Takahashi, K.(2006).Inhibition properties of dipeptides from salmon muscle hydrolysae on angiotensin I-converting enzyme.Int. J. Food Sci. Technol.,41,383-386.
33. Oshima, G.,Shimabukuro, H.,Nagasawa, K.(1979).Peptide inhibitors of angiotensin I-converting enzyme in digests of gelatin by bacterial collagenase.Biochim. Biophys. Acta,566,128-137.
34. Saiga, A.,Okumura, T.,Makihara, T.,Katsuda, S. I.,Morimatsu, F.,Nishimura, T.(2006).Action mechanism of an angiotensin I-converting enzyme inhibitory peptide derived from chicken breast muscle.J. Agric. Food Chem.,54,942-945.
35. Seki, E.,Osajima, K.,Matsufuji, H.,Matsui, T.,Osajima, Y.(1995).Val-Tyr, an angiotensin-I converting enzyme-inhibitor from sardines that have resistance to gastrointestinal proteases.J. Jpn. Soc. Biosci. Biotechol. Agrochem.,69,1013-1020.
36. Sentandreu, M. A.,Toldra, F.(2007).Evluation of ACE inhibitory activity of dipeptides generatd by the action of porcine muscle dipeptidyl peptidases.Food Chem.,102,511-515.
37. Suetsuna, H.(1998).Purification and identification of angiotensin I-converting enzyme inhibitors from the red alga (Porphyra yezoensis).J. Mar. Biotechnol,6,163-167.
38. Suetsuna, K.,Chen, J. R.(2001).Identification of antihypertensive peptides from peptic digests of two microalgae, Chlorella vulgaris and Spirulina platensis.J. Mar. Biotechnol.,3,305-309.
39. Tauzin, J.,Miclo, L.,Gaillard, J. L.(2002).Angiotensin-I-converting enzyme inhibitort peptides frum tryptic hydrolysate of bovine αs2-casein.FEBS Lett.,531,369-374.
40. Turner, A. J.,Hooper, N. M.(2002).The angiotensin-converting enzyme gene family: genomics and pharmacology.Trends Pharmacol. Sci.,23,177-183.
41. Yang, H. Y. T.,Erdos, E. G.,Levin, Y.(1970).A dipeptidyl carboxypeptidase that converts angiotensin II and inactivates bradykinin.Biochi. Biophys. Acta,214,374-376.
42. Yang, Y.,Marczak, E. D.,Yokoo, M.,Usui, H.,Yoshikawa, M.(2003).Isolation and antihypertensive effect of angiotensin I-converting enzyme inhibitory peptides from spinach Rubisco.J. Agri. Food Chem.,51,4897-4902.
43. Yokoyama, K. H.,Chiba, H.,Yoshikawa, M.(1992).Peptide inhibitors for angiotensin-I-converting enzyme from thermolysin digest of dried bonito.Biosci. Biotechnol. Biochem,56(6),1541-1545.