扇贝裙边ACE抑制肽的分离鉴定及其降血压活性研究
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摘要
为研究扇贝裙边ACE抑制肽的结构及其降血压效果,以虾夷扇贝Patinopecten yessoensis裙边酶解液为研究对象,采用凝胶色谱及反相高效液相色谱对ACE抑制肽组分进行分离纯化、ESI MS/MS鉴定和动物试验。结果表明:经Sephadex LH-20分离得到的组分F5和F7具有ACE抑制活性,其半抑制浓度(IC_(50))分别为1.4、1.7 mg/m L;通过两次反相高效液相色谱分离得到F5-3-3,经质谱解析筛选出的肽段Val-Trp(VW)具有ACE抑制活性,其IC_(50)为86.9μmol/L;用扇贝酶解液经超滤获得的相对分子量小于3000的组分灌胃雄性原发性高血压大鼠(SHR)28 d,灌胃第9天时,SHR的心脏收缩压(SBP)达到172 mm Hg,之后心脏收缩压基本保持恒定且极显著低于空白对照组(P<0.01),停止灌胃7 d内,多肽处理组SHR的心脏收缩压与空白对照组相比仍表现出极显著性差异(P<0.01)。研究表明,扇贝裙边酶解产物具有较好的降血压效果,可作为研发降压药物或保健食品的良好原料。
The angiotensin convert enzyme peptides(ACE)inhibitor peptides were purified from enzymatic hydrolysate of soft part excluding adductors of yesso scallop Patinopecten yessoensis by gel filtration chromatography and reversed-phase high-performance liquid chromatography(RP-HPLC),and then the structure and hypertensive activity of the ACE inhibitor peptides were studied.The fractions F5 and F7 separated by Sephadex LH-20 were shown to inhibit ACE,with 50%inhibitory concentration(IC_(50))of 1.4 mg/m L for F5 and 1.7 mg/m L for F7.The F5-3-3 was derived from F5-3 with RP-HPLC for twice,and the peptide Val-Trp(VW)identified by mass spectrometry(MS)had ACE inhibition,with IC_(50)of 86.9μmol/L.Then the enzymic hydrolysate with molecular weight of below 3000 obtained by filtration was gavaged to spontaneously hypertensive rat(SHR)for 28 days when very significantly decrease in systolic blood pressure(SBP)was observed in SHR(P<0.01),even very significantly lower(systolic pressure of 172 mm Hg)than that in the control group 9 days after intragastric administration(P<0.01).The findings indicate that the enzymic hydrolysate from soft part excluding adductors of yesso scallop has a role in reduction in blood pressure and becomes a candidate for development of hypoglycemic drugs and health food.
The angiotensin convert enzyme peptides(ACE)inhibitor peptides were purified from enzymatic hydrolysate of soft part excluding adductors of yesso scallop Patinopecten yessoensis by gel filtration chromatography and reversed-phase high-performance liquid chromatography(RP-HPLC),and then the structure and hypertensive activity of the ACE inhibitor peptides were studied.The fractions F5 and F7 separated by Sephadex LH-20 were shown to inhibit ACE,with 50%inhibitory concentration(IC_(50))of 1.4 mg/m L for F5 and 1.7 mg/m L for F7.The F5-3-3 was derived from F5-3 with RP-HPLC for twice,and the peptide Val-Trp(VW)identified by mass spectrometry(MS)had ACE inhibition,with IC_(50)of 86.9μmol/L.Then the enzymic hydrolysate with molecular weight of below 3000 obtained by filtration was gavaged to spontaneously hypertensive rat(SHR)for 28 days when very significantly decrease in systolic blood pressure(SBP)was observed in SHR(P<0.01),even very significantly lower(systolic pressure of 172 mm Hg)than that in the control group 9 days after intragastric administration(P<0.01).The findings indicate that the enzymic hydrolysate from soft part excluding adductors of yesso scallop has a role in reduction in blood pressure and becomes a candidate for development of hypoglycemic drugs and health food.
引文
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[19]阮氏香江,陈宁,黄婉苏,等.鬼针草水提物的降血压作用及其作用机制研究[J].广西医科大学学报,2017,34(2):177-180.
[20]邵江娟,李蒙蒙,付金柏.牡蛎配伍葛根对自发性高血压(雄性原发性高血压大鼠)大鼠降血压作用机制研究[J].江苏中医药,2017,49(8):75-78.
[21]全向阳,刘芸,甘海宁.水解酪蛋白肽粉复方对高血压大鼠的降血压作用及机制研究[J].保健医学研究与实践,2016,13(2):43-46,57.
[2]吕青,陈彦兵,马云杰.我国贝类出口遭遇的技术性贸易壁垒及应对措施[J].安徽农业科学,2007,35(35):11655-11657.
[3]农业部渔业渔政管理局.中国渔业统计年鉴2016[M].北京:中国农业出版,2016:29-33.
[4]丁琦.扇贝裙边发酵调味品开发研究[D].大连:大连工业大学,2008.
[5]徐文超.扇贝蛋白的酶降解研究[D].青岛:中国海洋大学,2015.
[6] Cudennec B,Violle N,ChataignéG,et al.Evidence for an antihypertensive effect of a land snail(Helix aspersa)by-product hydrolysate-identification of involved peptides[J].Journal of Functional Foods,2016,22:602-611.
[7] Rawendra R D S,Aisha S,Chang C I,et al.A novel angiotensin converting enzyme inhibitory peptide derived from proteolytic digest of Chinese soft-shelled turtle egg white proteins[J].Journal of Proteomics,2013,94(20):359-369.
[8] Lin lin,Lv Shun,Li Bafang. Angiotensin-I-converting enzyme(ACE)-inhibitory and antihypertensive properties of squid skin gelatin hydrolysates[J].Food Chemistry,2012,131(1):225-230.
[9] Bougatef A,Nedjar-Arroume N,Ravallec-PléR,et al.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[J].Food Chemistry,2008,111(2):350-356.
[10]刘媛.海湾扇贝酶解物的制备、鉴定及其体内外抗肿瘤作用研究[D].保定:河北农业大学,2015.
[11]李荣乔.菠萝蛋白酶酶解海湾扇贝裙边制备抗氧化肽的研究[D].保定:河北农业大学,2014.
[12]肖斌.鲍鱼内脏蛋白质酶解制备降血压多肽的研究[D].福州:福建农林大学,2013.
[13] Wang Jiapei,Hu Jianen,Cui Jinzhe,et al.Purification and identification of a ACE inhibitory peptide from oyster proteins hydrolysate and the antihypertensive effect of hydrolysate in spontaneously hypertensive rats[J]. Food Chemistry,2008,111(2):302-308.
[14]赵玉菲,顾伟,徐永健,等.大海马ACE抑制肽的制备及其理化性质分析[J].中国食品学报,2018,18(4):139-147.
[15]华鑫,孙乐常,万楚君,等.刺参ACE抑制肽制备及降压功效分析[J].食品科学,2018,39(10):125-130.
[16]王晶晶,吉薇,吉宏武,等.远东拟沙丁鱼酶解产物的降血压效果评价及其ACE抑制肽的分离纯化与鉴定[J].食品与发酵工业,2016,42(4):63-68.
[17]张绵松,史亚萍,袁文鹏,等.青柳蛤酶解条件优化及辅助降血压活性的研究[J].食品工业,2017,38(3):52-56.
[18]吴秉宇,张建华,左琦.羧肽酶A/B水解港湾扇贝裙边分离ACE抑制肽[C]//中国食品科学技术学会.北京:中国食品科学技术学会,2014.
[19]阮氏香江,陈宁,黄婉苏,等.鬼针草水提物的降血压作用及其作用机制研究[J].广西医科大学学报,2017,34(2):177-180.
[20]邵江娟,李蒙蒙,付金柏.牡蛎配伍葛根对自发性高血压(雄性原发性高血压大鼠)大鼠降血压作用机制研究[J].江苏中医药,2017,49(8):75-78.
[21]全向阳,刘芸,甘海宁.水解酪蛋白肽粉复方对高血压大鼠的降血压作用及机制研究[J].保健医学研究与实践,2016,13(2):43-46,57.