猪血红蛋白ACE抑制肽的分离和理化性质研究
详细信息Study on separation and physicochemical properties of ACE inhibitory peptides from porcine hemoglobin
-
摘要: 以猪血红蛋白的胃蛋白酶水解液为原料,依次通过大孔吸附树脂DA201-C、葡聚糖凝胶SephadexLH-20进行纯化,最终得到了3个降血压组分;并收集ACE抑制活性最高的组分α-Ⅱ,对其进行理化性质的研究。结果表明,酶解液经过大孔树脂DA201-C脱盐后,IC50为0.87mg/mL;继续经过葡聚糖凝胶SephadexLH-20分离得到的猪血红蛋白ACE抑制肽（α-Ⅱ）的IC50为0.21mg/mL。分离后的猪血红蛋白ACE抑制肽（α-Ⅱ）在pH为7时,溶解度最低,为50.37%;胃蛋白酶对其活性影响不显著。温度在25～55℃之间以及中性和偏酸性的条件下时,α-Ⅱ具有较好的热稳定性;当NaCl浓度大于0.6mol/L时,其活性急剧下降。Abstract: The pepsin hydrolysate of porcine hemoglobin as raw material was used for adsorption and desorption through using macroporous resin DA201-C and Sephadex LH-20 chromatogram, then three antihypertensive components were obtained;and to collect and to study on physicochemical properties of ACE inhibitory peptideα-Ⅱthat possessing the largest ACE inhibitory activity.The results showed that:ACE inhibitory peptide of porcine hemoglobin was separaed by macroporous resin DA201-C and the IC50 was 0.87mg/mL;next separaed by Sephadex LH-20 and IC50 was 0.21mg/mL.The lowest solubility of ACE inhibitory peptides of porcine hemoglobin (α-Ⅱ) was 50.37%at pH 7;and this fraction (α-Ⅱ) demonstrated high stability against gastrointestinal proteases, temperature 25～55℃and pH<7.But ACE inhibitory activity could reduce rapidly when the NaCl concentration exceeded 0.6mol/L.
-
[1] MURRAY B A, FITZGERALD R J.Angiotensin converting enzyme inhibitory peptides derived from food proteins:Biochemistry, bioactivity and production[J].Current Pharmaceutical Design, 2007, 13 (8) :773-791.
[2] CHEUNG I W, NAKAYAMA S, MONICA N K, et al.Angiotensin-I converting enzyme inhibitory activity of hydrolysates from oat (Avena sativa) proteins by in silico and in vitro analyses[J].Journal of Agricultural and Food Chemistry, 2009, 57 (19) :9234-9242.
[3] FERREIRA S H.A bradykinin-potentiating factor (BPF) present in the venom of Bothrops jararaca[J].British Journal of Pharmacology and Chemotherapy, 1965, 24 (1) :163-169.
[4] JIMSHEENA V K, GOWDA R L.Angiotensin I-converting enzyme (ACE) inhibitory peptides derived from arachin by simulated gastric digestion[J].Food Chemistry, 2011, 125 (2) :561-569.
[5] PIHLANTOLEPPALA A, ROKKA T, KORHONEN H.Angiotensin I converting enzyme inhibitory peptides derived from bovine milk proteins[J].International Dairy Journal, 1998, 8 (4) :325-331.图4pH对猪血红蛋白ACE抑制肽 (α-Ⅱ) 的ACE抑制活性的影响Fig.4Effect of pH value on the activity of ACE inhibitory peptide (α-Ⅱ) from porcine hemoglobin [6] FUJITA H, YOSHIKAWA M.LKPNM:a prodrug-type ACE-inhibitory peptide derived from fish protein[J].Immunopharmacology, 1999, 44 (1/2) :123-127.
[7] Pan Dao-dong, Luo Yong-kang, TANOKURA M.Antihypertensive peptides from skimmed milk hydrolysate digested by cell-free extract of Lactobacillus helveticus JCM1004[J].Food Chemistry, 2005, 91 (1) :123-129.
[8] Zhao Yuan-hui, Li Ba-fang, Liu Zun-ying, et al.Antihypertensive effect and purification of an ACE inhibitory peptide from sea cucumber gelatin hydrolysate[J].Process Biochemistry, 2007, 42 (12) :1586-1591.
[9] Wang Jia-pei, Hu Jia-nen, Cui Jin-zhe, et al.Purification and identification of a ACE inhibitory peptide from oyster proteins hydrolysate and the anti hypertensive effect of hydrolysate in spontaneously hypertensive rats[J].Food Chemistry, 2008, 111 (2) :302-308.
[10] BYUN H G, KIM S K.Purification and characterization of angiotensin I converting enzyme (ACE) inhibitory peptides from Alaska pollack (Theragra chalcogramma) skin[J].Process Biochemistry, 2011, 36 (12) :1155-1162.
[11] Han Hua, Yi Yang-hua, Li Ling, et al.A new triterpene glycoside from sea cucumber Holothuria leucospilota[J].Chinese Chemical Letters, 2007, 18 (2) :161-164.
[12] KIM M R, KAWAMURA Y, LEE C H.Isolation and identification of bitter peptides of tryptic hydrolysate of soybean11S glycinin by reverse-phase high-performance liquid chromatography[J].Journal of Food Science, 2003, 68 (8) :2416-2422.
[13] Wu Jiang-ping, Ding Xiao-lin.Characterization of inhibition and stability of soy-protein-derived angiotensinⅠ-converting enzyme inhibitory peptides[J].Food Research International, 2002, 35 (4) :367-375.
[14] TAKUO N, LECH O.Purification of glycomacropeptide from nondialyzable fraction of sweet whey by hydrophobic interaction chromatography on phenyl-agarose[J].Biotechnology Letters, 2000, 22 (5) :413-416.
[15] ALEMAN A, GIMENEZ B, PEREZ-SANTIN E, et al.Contribution of Leu and Hyp residues to antioxidant and ACE-inhibitory activities of peptide sequences isolated from squid gelatin hydrolysate[J].Food Chemistry, 2011, 125 (2) :334-341.
[16] Zhao Qiu-yu, SANNIER F, PIOT J M.Kinetics of appearance of four hemorphins from bovine hemoglobin peptic hydrolysates by HPLC coupled with photodiode array detection[J].Biochimica et Biophysica Acta (BBA) -Protein Structure and Molecular Enzymology, 1996, 1295 (1) :73-80.
[17] Wan J B, Zhang Q W, Ye W C, et al.Quantification and separation of protopanaxatriol and protopanaxadiol type saponins from Panax notoginseng with macroporous resins[J].Separation and Purification Technology, 2008, 60 (2) :198-205.
[18] 刘佳.大豆蛋白ACE抑制肽的研究[D].无锡:江南大学, 2008. [19] RIIHINEN K R, GODECKE T, PAULI G F, et al.Purification of berry flavonol glycosides by long-bed gel permeation chromatography[J].Journal of chromatography A, 2012, 1244 (29) :20-27.
[20] Wu J, ALUKO R, MUIR A.Imp roved Method for Direct High-performance Liquid Chromatography Assay of Angiotensin-converting Enzyme-catalyzed Reaction[J].Journal of Chromatograpy A, 2002, 950:125-131.
[21] 盛彩虹, 刘晔, 刘大川, 等.紫苏分离蛋白功能性研究[J].食品科学, 2011, 32 (17) :137-139. [22] 陈季旺, 孙勤, 夏文水.鱼降压肽的大孔树脂分离及其活性稳定性[J].食品科学, 2009, 30 (18) :25-28. [23] LIESELOT V, GUY S, TOSHIRO M, et al.C (ACE) inhibitory peptide from the gastrointestinal hydrolysate of the cotton leafworm, Spodoptera littoralis[J].Process Biochemistry, 2008, 43 (8) :900-904.
[24] 廖丹葵.鸡蛋蛋黄蛋白质制备降血压肽的研究[D].南宁:广西大学, 2006. [25] 杨锋, 马千里, 黄永春.醋蛋中ACE抑制肽的分离及稳定性研究[J].食品科学, 2009, 30 (24) :78-79. [26] Jiang Jiang, Chen Jie, Xiong You-ling.Structural and emulsifying properties of soy protein isolate subjected to acid and alkaline pH-shifting processes[J].Journal of Agricultural and Food Chemistry, 2009, 57 (16) :7576-7583.
[27] 潘道东, 林璐.DA201-C大孔吸附树脂静态吸附ACE抑制肽的研究[J].食品科学, 2009, 30 (5) :20-23.
计量
- 文章访问数: 94
- HTML全文浏览量: 6
- PDF下载量: 144
下载:
