花生ACE抑制肽结构表征与构效关系

doi:10.7506/spkx1002-6630-201309035

食品科学 ›› 2013, Vol. 34 ›› Issue (9): 170-174.doi: 10.7506/spkx1002-6630-201309035

花生ACE抑制肽结构表征与构效关系

王强1，王春艳1,2 ，胡晖1，刘红芝1，刘丽1

1.中国农业科学院农产品加工研究所，农业部农产品加工综合性重点实验室，北京 100193；
2.沈阳农业大学食品学院，辽宁沈阳 110866

收稿日期:2013-03-12 修回日期:2013-04-16 出版日期:2013-05-15 发布日期:2013-05-07
通讯作者: 王强 E-mail:wangqiang365@263.net
基金资助:
农业部公益性行业科研专项(201203037)；国家国际科技合作专项(2012DFA31400)

Structural Characterization and Structure-activity Relationship of ACE Inhibitory Peptides from Peanut

WANG Qiang1，WANG Chun-yan1,2，HU Hui1，LIU Hong-zhi1，LIU Li1

1. Key Laboratory of Agro-products Processing, Ministry of Agriculture, Institute of Agro-products Processing Science and
Technology, Chinese Academy of Agricultural Sciences, Beijing 100193, China；
2. College of Food Science, Shenyang Agricultural University, Shenyang 110866, China

Received:2013-03-12 Revised:2013-04-16 Online:2013-05-15 Published:2013-05-07
Contact: Qiang Wang E-mail:wangqiang365@263.net

摘要/Abstract

摘要：

在碱性蛋白酶Alcalase与N120P蛋白酶复合酶解花生分离蛋白制备花生短肽的基础上，采用凝胶过滤色谱与制备型反相高效液相色谱(RP-HPLC)分离纯化得到27个组分，从中筛选出ACE抑制活性最高的组分P8，其在0.010mg/mL质量浓度条件下ACE抑制率达85.77%，IC50值为0.0052mg/mL(6.42μmol/L)。采用基质辅助激光解吸附电离飞行时间串联质谱(MALDI-TOF-TOF MS)对P8的氨基酸序列进行分析，得出P8的相对分子质量为808.80，氨基酸序列为Lys-Leu-Tyr-Met-Arg-Pro(KLYMRP)。通过固相合成的方法合成短肽KLYMRP，并证实其具有显著的ACE抑制活性，ACE抑制IC50值为0.0038mg/mL(4.69μmol/L)。构效关系分析结果表明：短肽Lys-Leu-Tyr-Met-Arg-Pro的C末端为Pro，N末端为碱性氨基酸Lys，且肽的疏水性很强，有利于与ACE活性部位的结合；短肽KLYMRP与ACE活性位点的残基形成5个氢键，一个Pi-Cation相互作用，并与锌离子形成配位键，这些作用力共同稳定了短肽-ACE复合物的构象，从而有利于ACE抑制活性的发挥。

关键词: 花生短肽, 氨基酸序列, ACE抑制活性, 分子构象, 构效关系

Abstract:

Peanut short-chain peptides were prepared by hydrolyzing peanut protein isolate with both alcalase and protease
N120P and separated by gel filtration chromatography and preparatve RP-HPLC. As a result, 27 components were obtained,
and P8 was found to have the highest ACE inhibitory activiy among them, with an inhbitory rate of 85.77% at 0.010 mg/mL
and an IC50 of 0.0052 mg/mL (6.42 μmol/L). The relativel molecular mass of the component P8 was 808.80 with an amino
acid sequence of Lys-Leu-Tyr-Met-Arg-Pro(KLYMRP) as determined by MALDI-TOF-TOF MS. The short-chain peptide
KLYMRP prepared with P8 by solid-phase synthesis was confirmed to have potent ACE inhibitory activity with an IC50 of
0.0038 mg/mL (4.69 μmol/L). Analysis of structure-activity relationship indicated that the short-chain peptide Lys-Leu-Tyr-
Met-Arg-Pro consisted of Pro at the C terminus and Lys at the N terminus and had strong hydrophobicity, thus facilitating
active site docking with ACE. In addition, KLYMRP formed five hydrogen bonds and one Pi-Cation interaction with residual
groups at the ACE active positions and formed coordination bonds with Zn ions. All these contributed greatly to stabilize the
conformation of the ACE-KLYMRP complex and therefore benefited ACE inhibition.

Key words: peanut peptides, amino acid sequence, ACE inhibitory activity, molecular conformation, structure-activity relationship

中图分类号:

TQ936.16

王强1，王春艳1,2,胡晖1，刘红芝1，刘丽1. 花生ACE抑制肽结构表征与构效关系[J]. 食品科学, 2013, 34(9): 170-174.

WANG Qiang1，WANG Chun-yan1,2，HU Hui1，LIU Hong-zhi1，LIU Li1. Structural Characterization and Structure-activity Relationship of ACE Inhibitory Peptides from Peanut[J]. FOOD SCIENCE, 2013, 34(9): 170-174.

参考文献

[1]张宇昊, 王强.花生蛋白的开发与利用[J].花生学报,2005,34(4):12-16

[2]柳杰. 花生粕抗氧化肽的研究[D]. 无锡: 江南大学, 2010.

[3]Su G W, Cui C, Zheng L, et al.Isolation and identification of two novel umami and umami-enhancing peptides from peanut hydrolysate by consecutive chromatography and MALDI-TOF/TOF MS[J].Food Chemistry,2012,135:479-485

[4]Zaliani A, Gancia E J.Structure and activity of angiotensin I converting enzyme inhibitory peptides from sake and sake lees[J].Chemical Information and Modeling,1999,39:525-533

[5]刘焕, 乐国伟, 施用晖, 等.血管紧张素转化酶二肽抑制剂的构效关系[J].计算机与应用化学,2005,22(8):631-635

[6]黎观红, 乐国伟, 施用晖.花生分离蛋白水解物中血管紧张素转化酶抑制肽的分离纯化与结构鉴定[J].中国粮油学报,2010,25(6):56-61

[7]王瑛瑶, 王璋, 陈尚卫.花生ACE抑制活性肽的分离纯化与结构鉴定[J].食品科学,2008,29(10):341-344

[8]Wang N F, Li C Y, Yan Zheng.Purification and characterization of angiotensin I-converting enzyme inhibitory peptide from lactic acid fermented peanut flour[J].Journal of Biotechnology,2008,136:717-742

[9]Quist E E, Phillips R D, Saalia F K.Angiotensin converting enzyme inhibitory activity of proteolytic digests of peanut (Arachis hypogaea L) flour[J]. Food Science and Technology,2009,42(3):6-

[10]Guang C, Phillips R D.Purification, Activity and Sequence of Angiotensin I Converting Enzyme Inhibitory Peptide from Alcalase Hydrolysate of Peanut Flour[J].Journal of Agricultural and Food Chemistry,2009,57(21):10102-10106

[11]Eresha M, Niranjan R, Se K K.Antioxi-dant properties of a radical-scavenging peptide purified from enzymatically prepared fish skin gelatin hydrolysate[J].Journal of Agricultural＆Food Chemistry,2005,53(3):581-587

[12]Suetsuna K.nullUkeda H, Ochi H. Isolation and characterization of freeradical scavenging activities peptides derived from casein[J]. Journal of Nutritional Biochemistry,2000,:-

[13]Rajapakse N.nullMendis E, Byun H G, et a1. Purification and invitro antioxidative effects of giant squid muscle peptides on free radical—mediated oxidative systems[J].. Journal of Nutrition Biochemistry,2005,:-

[14]Qian Z J, Jung W K, Kim S K.Free radical scavenging activity of a novel antioxidative peptide purified from hydrolysate of bullfrog skin，Rana catesbeiana Shaw[J].Bioresource Technology,2008,99(6):1690-

[15]Takahashi N, Togashi S, Watanahe S S, et a1.Antioxidatie collagen-derived peptides in human-placenta extract[J].Placenta,2002,(23):497-502

[16]Fitz R J, Meisel H.Milk protein-derived peptide inhibitors of angiotensin I-converting enzyme[J].British Journal of Nutrition,2000,84(Supp11):33-37

[17]陈芳进, 刘涛, 潘和平, 等.N端6-甲基辛酰基化修饰对小肽KTKKKFLKKT结构的影响[J].自然科学进展,2009,19(10):1056-1061

[18]郭慧青, 毛惠, 赵波, 等.两种血管紧张素转化酶抑制肽作用于靶标的分子机理[J].食品科学,2010,31(23):1-5

[19]Pan D D, Cao J X, Guo H Q, et al.Studies on purification and the molecular mechanism of a novel ACE inhibitory peptide from whey protein hydrolysate[J].Food Chemistry,2012,1(1):121-126

[20]David W O.[J].Norman H N. Principles of modern chemistry[M]. New York: Saunders College Publishing,1986,:-

花生ACE抑制肽结构表征与构效关系

Structural Characterization and Structure-activity Relationship of ACE Inhibitory Peptides from Peanut

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