南瓜籽蛋白源血管紧张素转化酶抑制肽的制备及其降血压活性

doi:10.7506/spkx1002-6630-20201118-197

食品科学 ›› 2021, Vol. 42 ›› Issue (23): 214-220.doi: 10.7506/spkx1002-6630-20201118-197

南瓜籽蛋白源血管紧张素转化酶抑制肽的制备及其降血压活性

何海艳，刘梦婷，杨爱萍，蒋彩云，崔逸，汪洪涛

（1.江苏经贸职业技术学院健康学院，江苏?南京 211168；2.南京财经大学食品科学与工程学院，江苏?南京 210023）

出版日期:2021-12-15 发布日期:2021-12-29
基金资助:
江苏经贸职业技术学院校级重点课题（JSJM009）；江苏省高等学校自然科学研究面上项目（17KJB550002）；江苏省高校优秀科技创新团队项目

Preparation of Angiotensin Converting Enzyme Inhibitory Peptides from Pumpkin Seed Protein and Their Antihypertensive Activity

HE Haiyan, LIU Mengting, YANG Aiping, JIANG Caiyun, CUI Yi, WANG Hongtao

(1. School of Health, Jiangsu Vocational Institute of Commerce, Nanjing 211168, China;2. College of Food Science and Engineering, Nanjing University of Finance & Economics, Nanjing 210023, China)

Online:2021-12-15 Published:2021-12-29

摘要/Abstract

摘要： 食源性降血压肽在调节机体血压过程中发挥着一定的积极作用。本实验采用碱性蛋白酶水解南瓜籽蛋白，水解物经膜分离获得血管紧张素转化酶（angiotensin converting enzyme，ACE）抑制肽，通过质谱鉴定其结构，并采用抑制动力学和分子对接方法研究ACE抑制肽的活性机制；此外，通过ACE抑制活性实验、自发性高血压大鼠（spontaneously hypertensive rats，SHRs）模型等评价南瓜籽蛋白水解物、膜分离组分以及南瓜籽肽的降血压活性。结果表明，低于1 kDa南瓜籽蛋白水解物组分ACE抑制活性较好，1 mg/mL下ACE抑制率为46.22%，100 mg/kg mb 剂量下灌胃SHRs 6 h后收缩压可以降低21.42 mm Hg；鉴定出9 个ACE抑制肽（LLV、LVF、LTPL、SVLF、LLPQ、MLPL、LLPGF、VLLPE和RFPLL），其中RFPLL、LLPGF、MLPL和LVF具有较好的ACE抑制活性，半抑制浓度均低于1 mmol/L；RFPLL和LVF具有较好的降血压活性，30 mg/kg mb剂量下灌胃6 h后SHRs的收缩压降低量分别为37.0 mm Hg和22.2 mm Hg，SHRs的舒张压降低量分别为17.0 mm Hg和11.2 mm Hg；RFPLL、LLPGF、MLPL和LVF可以很好地对接ACE的活性中心，RFPLL是ACE混合型抑制剂，MLPL对ACE的抑制模式可能是竞争型抑制，LLPGF、LVF可能是ACE的非竞争型抑制剂。综上，低于1 kDa南瓜籽蛋白水解物是一种良好的降血压功能食品原料，其中RFPLL和LVF可用于降血压功能食品或者药品的开发原料。

关键词: 南瓜籽肽；血管紧张素转化酶；降血压活性；抑制动力学；分子对接

Abstract: Food-derived antihypertensive peptides play a positive role in regulating blood pressure. In this study, angiotensin converting enzyme (ACE) inhibitory peptides from the hydrolysate of pumpkin seed protein with alcalase for 4 h was separated by membrane filtration and identified structurally by mass spectrometry (MS). The action mechanism of the ACE inhibitory peptides was studied by kinetic analysis of the inhibitory effect and molecular docking. In addition, ACE inhibition assay and spontaneously hypertensive rats (SHRs) were used to evaluate the antihypertensive activity of the protein hydrolysate, its fractions separated by membrane filtration and the peptides present in the fraction with molecular mass less than 1 kDa. The results showed that the fraction with molecular mass less than 1 kDa had good ACE inhibitory activity, with an inhibition rate of 46.22% at a concentration of 1 mg/mL. The systolic blood pressure of SHRs decreased by 21.42 mm Hg at 6 hours after oral administration with this faction at a dose of 100 mg/kg mb. Nine ACE inhibitory peptides were identified, namely LLV, LVF, LTPL, SVLF, LLPQ, MLPL, LLPGF, VLLPE and RFPLL, and RFPLL, LLPGF, MLPL and LVF had good ACE inhibitory activity, with a half maximal inhibitory concentration less than 1 mmol/L. RFPLL and LVF had relatively better antihypertensive activity, decreasing the systolic blood pressure of SHRs by 37.0 and 22.2 mm Hg, and decreasing the diastolic blood pressure by 17.0 and 11.2 mm Hg at 6 hours after gavage administration at a dose of 30 mg/kg mb, respectively. RFPLL, LLPGF, MLPL and LVF could dock at the active center of ACE very well. RFPLL was a mixed inhibitor of ACE. ACE could be inhibited by MLPL competitively, while LLPGF and LVF could be non-competitive inhibitors of ACE. Therefore, pumpkin seed protein hydrolysates with molecular mass less than 1 kDa are promising ingredients of functional foods with antihypertensive activity, and RFPLL and LVF can be used for the development of antihypertensive functional foods or drugs.

Key words: pumpkin seed peptide; angiotensin converting enzyme; antihypertensive activity; inhibitory kinetics; molecular docking

中图分类号:

TS255.1

何海艳，刘梦婷，杨爱萍，蒋彩云，崔逸，汪洪涛. 南瓜籽蛋白源血管紧张素转化酶抑制肽的制备及其降血压活性[J]. 食品科学, 2021, 42(23): 214-220.

HE Haiyan, LIU Mengting, YANG Aiping, JIANG Caiyun, CUI Yi, WANG Hongtao. Preparation of Angiotensin Converting Enzyme Inhibitory Peptides from Pumpkin Seed Protein and Their Antihypertensive Activity[J]. FOOD SCIENCE, 2021, 42(23): 214-220.

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南瓜籽蛋白源血管紧张素转化酶抑制肽的制备及其降血压活性

Preparation of Angiotensin Converting Enzyme Inhibitory Peptides from Pumpkin Seed Protein and Their Antihypertensive Activity

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