两种来源于牛皮胶原蛋白的新型ACE抑制肽

董莹; 王湛清; 沈菊泉; 范季瀛; 汤俊; 马志英; 沈亚领

doi:10.13386/j.issn1002-0306.2017.10.018

两种来源于牛皮胶原蛋白的新型ACE抑制肽

1. 华东理工大学生物工程学院,生物反应器工程国家重点实验室
2. 上海市食品研究所

基金项目:

国家重点实验室专项经费资助项目(2060204)；上海重点学科建设项目(B505)；

详细信息

作者简介:
董莹 (1983-) , 女, 硕士研究生, 研究方向:食品科学, E-mail:doney0211@126.com。;

沈亚领 (1960-) , 男, 博士, 教授, 研究方向:生物工程, E-mail:ylshen@ecust.edu.cn。;

中图分类号: TQ936.16
计量
- 文章访问数: 0130
- HTML全文浏览量: 019
- PDF下载量: 0119
出版历程
- 收稿日期: 2016-08-31

Two novel ACE inhibitory peptides derived from bovine skin collagen

State Key Laboratory of Bioreactor Engineering, School of Bioengineering, East China University of Science and Technology
Shanghai Food Research Institute

摘要

摘要: 从牛皮胶原蛋白中鉴定分离出抑制血管紧张素Ⅰ转换酶（ACE）的多肽片段。首先优化牛皮胶原蛋白水解方法,应用胃蛋白酶和碱性蛋白酶获得活性最高的水解产物（抑制ACE的IC50为0.168 mg/m L）。经MW 5000超滤分离后,得到水解液中活性最高的部分（IC50为0.079 mg/m L）。对分离后低于MW 5000的部分经RP-HPLC进一步分离,发现含有大量ACE抑制肽,并应用RP-HPLC-MS/MS鉴定出两种新型ACE抑制肽,氨基酸序列分别为ISVPGPM和LGPVGNPGPA,IC50值分别为0.017 mg/m L（24.3μmol/L）和0.077 mg/m L（87.8μmol/L）。最后,本文模拟了两种抑制肽在体内生理环境下的消化,发现均可被部分降解,且消化产物具有与原始多肽相近的ACE抑制活性。
- 牛皮胶原蛋白 /
- 水解产物 /
- ACE抑制肽
Abstract: Peptidic fractions which inhibit angiotensin I-converting enzyme ( ACE) were identified from bovine skin collagen.First, bovine skin collagen was hydrolyzed by an optimized method using pepsin and alcalase.Hydrolyzed collagen obtained the most active hydrolysate ( inhibitory against ACE with an IC50 of 0.168 mg/m L) . With an ultrafiltration by MW 5000, the most active fraction of the hydrolysate was obtained ( IC50 of 0.079 mg/m L) .After further separations with the fraction lower than MW5000 by RP-HPLC, considerable ACE inhibitory peptides were found. Two novel ACE inhibitory were identified by RP-HPLC-MS/MS. The amino acid sequences were ISVPGPM and LGPVGNPGPA, IC50 values of 0.017 mg/m L ( 24.3 μmol/L) and 0.077 mg/m L ( 87.8 μmol/L) , respectively.Finally, the digestion of the two peptides in vivo physiological environment was simulated and both of them were partly degraded. The ACE inhibitory activity of the digestive products were similar to the original polypeptides.
- bovine skin collagen /
- hydrolysate /
- ACE inhibitory peptide

HTML全文

参考文献(26)

[1]	Martinez M D, Miralles B, Recio I, et al.Antihypertensive peptides from food proteins:a review[J].Food&Function, 2012 (3) :350-361.
[2]	Natesh R, Schwager S L, Sturrock E D, et al.Crystal structure of the human angiotensin-converting enzyme-lisinopril complex[J].Nature, 2003 (421) :551-554.
[3]	Acharya K R, Sturrock E D, Riordan J F, et al.Ace revisited:A new target for structure-based drug design[J].Nature Reviews Drug Discovery, 2003 (2) :891-902.
[4]	Bhat Z F, Kumar S, Bhat H F.Antihypertensive Peptides of Animal Origin:A review[J].Critical Reviews in Food Science&Nutrition, 2017 (57) :566-578.
[5]	Hartmann R, Meisel H.Food-derived peptides with biological activity:from research to food applications[J].Current Opinion in Biotechnology, 2007 (18) :163-169.
[6]	Pripp A H.Initial proteolysis of milk proteins and its effect on formation of ACE-inhibitory peptides during gastrointestinal proteolysis:a bioinformatic, in silico, approach[J].European Food Research and Technology, 2005 (10) :1-5.
[7]	Quirós A, Ramos M, Muguerza B, et al.Identification of novel antihypertensive peptides in milk fermented with Enterococcus faecalis[J].International Dairy Journal, 2007 (17) :33-41.
[8]	Miguel M, Aleixandre M A, Ramos M, et al.Effect of simulated gastrointestinal digestion on the antihypertensive properties of ACE-inhibitory peptides derived from ovalbumin[J].Journal of Agricultural and Food Chemistry, 2006 (54) :726-731.
[9]	Hyun C K, Shin H K.Utilization of bovine blood plasma proteins for the production of angiotensin I converting enzyme inhibitory peptides[J].Process Biochemistry, 2000 (36) :65-71.
[10]	Li GH, Shi YH, Liu H, et al.Antihypertensive effect of alcalase generated mung bean protein hydrolysates in spontaneously hypertensive rats[J].Eur Food Res Technol, 2006 (222) :733-736.
[11]	Jung WK, Mendis E, Je JY, et al.Angiotensin I-converting enzyme inhibitory peptide from yellowfin sole (Limanda aspera) frame protein and its antihypertensive effect in spontaneously hypertensive rats[J].Food Chemistry, 2006 (94) :26-32.
[12]	Fahmi A, Morimura S, Guo H C, et al.Production of angiotensin I converting enzyme inhibitory peptides from sea bream scales[J].Process Biochemistry, 2004 (39) :1195-1200.
[13]	Zhen W, Pan D, Zhen X, et al.Angiotensin I-converting enzyme inhibitory peptides derived from bovine casein and identified by MALDI-TOF-MS/MS[J].Journal of the Science of Food&Agriculture, 2013 (93) :1331-1337.
[14]	Cushman D W, Cheung H S.Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung[J].Biochemical Pharmacology, 1971 (20) :1637-1648.
[15]	Gómez-Ruiz J, Taborda G, Amigo L, et al.Identification of ACE-inhibitory peptides in different Spanish cheeses by tandem mass spectrometry[J].Eur Food Res Technol, 2006 (223) :595-601.
[16]	Miguel M, Recio I, Gomez-Ruiz J A, et al.Angiotensin I-converting enzyme inhibitory activity of peptides derived from egg white proteins by enzymatic hydrolysis[J].Journal of Food Protection, 2004 (67) :1914-1920.
[17]	Chiang W D, Tsou M J, Tsai Z Y, et al.Angiotensin I-converting enzyme inhibitor derived from soy protein hydrolysate and produced by using membrane reactor[J].Food Chemistry, 2006 (98) :725-732.
[18]	Ruiz JáG, Ramos M, Recio I.Angiotensin converting enzyme-inhibitory activity of peptides isolated from Manchego cheese.Stability under simulated gastrointestinal digestion[J].International Dairy Journal, 2004 (14) :1075-1080.
[19]	Vermeirssen V, Camp J V, Decroos K, et al.The Impact of Fermentation and In Vitro Digestion on the Formation of Angiotensin-I-Converting Enzyme Inhibitory Activity from Pea and Whey Protein[J].Journal of Dairy Science, 2003 (86) :429-438.
[20]	Yu Y, Hu J, Miyaguchi Y, et al.Isolation and characterization of angiotensin I-converting enzyme inhibitory peptides derived from porcine hemoglobin[J].Peptides, 2006 (27) :2950-2956.
[21]	Alting A C, Meijer R J, Beresteijn E C V.Incomplete elimination of the ABBOS epitope of bovine serum albumin under simulated gastrointestinal conditions of infants[J].Diabetes Care, 1997 (20) :875-880.
[22]	Oomen A G, Hack A, Minekus M, et al.Comparison of five in vitro digestion models to study the bioaccessibility of soil contaminants[J].Environmental Science&Technology, 2002 (36) :3326-3334.
[23]	Cheung H S, Wang F L, Ondetti M A, et al.Binding of peptide substrates and inhibitors of angiotensin-converting enzyme.Importance of the COOH-terminal dipeptide sequence[J].The Journal of Biological Chemistry, 1980 (255) :401-407.
[24]	Shoji A, Yanagida A, Shindo H, et al.Counter-current chromatographic estimation of hydrophobicity of Z- (cis) and E- (trans) enalapril and kinetics of cis/trans isomerization[J].Journal of Chromatography A, 2007 (1157) :101-107.
[25]	Kim H M, Shin D R, Yoo O J, et al.Crystal structure of Drosophila angiotensin I-converting enzyme bound to captopril and lisinopril[J].FEBS Letters, 2003 (538) :65-70.
[26]	Gomez-Ruiz J A, Recio I, Belloque J.ACE-inhibitory activity and structural properties of peptide Asp-Lys-Ile-HisPro[beta-CN f (47-51) ].Study of the peptide forms synthesized by different methods[J].Journal of Agricultural And Food Chemistry, 2004 (52) :6315-6319.