食品科学 ›› 2025, Vol. 46 ›› Issue (1): 40-48.doi: 10.7506/spkx1002-6630-20231219-160

• 生物工程 • 上一篇    下一篇

鳖蛋黄血管紧张素转化酶抑制肽分离纯化及活性分析

刘华宇,廖彭莹,张天丰,张新锐,邓纭宁,李耀华,韦金锐,陈俊   

  1. (1.广西中医药大学药学院,广西 南宁 530200;2.广西中医药大学 广西高校中药提取纯化与质量分析重点实验室,中药学国家级实验教学示范中心,广西 南宁 530200;3.广西中医药大学 教学实验实训中心,广西 南宁 530200;4.广西中医药大学 广西中医药科学实验中心,广西 南宁 530200)
  • 出版日期:2025-01-15 发布日期:2024-12-30
  • 基金资助:
    国家自然科学基金地区科学基金项目(81960698); 广西高等学校千名中青年骨干教师培育计划项目(桂教师范[2019]81号); 中药学广西一流学科建设项目(桂教科研[2022]1号); 2022年自治区中药学研究生联合培养基地开放项目(桂学位[2021]6号); 广西壮瑶药重点实验室科研任务项目(GXZYKF2022-10); 广西中医药大学“桂派中医药传承创新团队”资助项目(2022A005); 2023年度广西高校中青年教师科研基础能力提升项目(2023KY0289)

Separation, Purification, and Activity Analysis of Angiotensin Converting Enzyme Inhibitory Peptides in Enzymatically Hydrolyzed Egg Yolk of Chinese Soft-Shelled Turtle

LIU Huayu, LIAO Pengying, ZHANG Tianfeng, ZHANG Xinrui, DENG Yunning, LI Yaohua, WEI Jinrui, CHEN Jun   

  1. (1. College of Pharmacy, Guangxi University of Chinese Medicine, Nanning 530200, China; 2. Key Laboratory of TCM Extraction and Purification and Quality Analysis, National Demonstration Center for Experimental Traditional Chinese Pharmacology, Guangxi University of Chinese Medicine, Nanning 530200, China; 3. Teaching Experiment and Training Centre, Guangxi University of Chinese Medicine, Nanning 530200, China; 4. Guangxi Scientific Research Centre of Traditional Chinese Medicine, Guangxi University of Chinese Medicine, Nanning 530200, China)
  • Online:2025-01-15 Published:2024-12-30

摘要: 为从鳖蛋黄酶解物中筛选具有血管紧张素转化酶(angiotensin converting enzyme,ACE)抑制活性的肽段,以ACE抑制活性为评价指标,采用超滤和凝胶过滤色谱技术进行分离纯化。采用液相色谱-串联质谱技术对活性组分进行肽段鉴定,借助生物信息学工具进行活性评估。优选预测活性较高的肽段进行合成和活性验证,并用分子对接工具分析活性肽与ACE的相互作用。结果表明,鳖蛋黄菠萝蛋白酶酶解物的水解度为(17.70±0.34)%,抑制ACE的半抑制浓度(half maximal inhibitory concentration,IC50)值为(0.210±0.019)mg/mL。对酶解产物进行分离纯化,从活性组分F3中鉴定出36 条肽段,选择6 条活性评分较高的肽段进行合成,其中肽段YNGIWPRD和ASDILPKK的IC50值分别为(0.019 00±0.000 36)、(0.170 0±0.001 3)mg/mL。分子对接结果表明,二者均通过多条氢键与ACE紧密结合。综上,从鳖蛋黄中筛选出2 条新的ACE抑制活性肽。

关键词: 鳖蛋黄;血管紧张素转化酶抑制肽;分离纯化;生物信息学;分子对接

Abstract: Angiotensin converting enzyme (ACE) inhibitory peptides were separated and purified from an enzymatic hydrolysate of egg yolk from Chinese soft-shelled turtle (Pelodiscus sinensis) by sequential ultrafiltration and gel filtration chromatography. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was utilized for the identification of the active peptides, and bioinformatics tools were utilized for activity evaluation. The peptides with strong ACE inhibitory activity were synthesized and verified, and molecular docking was used to analyze the interaction between the active peptides and ACE. The results showed that the hydrolysis degree of soft-shelled turtle egg yolk by bromelain was (17.70 ± 0.34)%, and the half maximal inhibitory concentration (IC50) of ACE inhibitory activity was (0.210 ± 0.019) mg/mL. Altogether, 36 peptides were identified from fraction F3 with ACE inhibitory activity. Six peptides with higher activity scores were selected for synthesis and activity verification, among which peptides YNGIWPRD and ASDILPKK exhibited strong ACE inhibitory activities with IC50 of (0.019 00 ± 0.000 36) and (0.170 0 ± 0.001 3) mg/mL, respectively. The molecular docking results showed that both peptides bounded to ACE tightly through multiple hydrogen bonds. In conclusion, two new ACE inhibitory peptides were selected from the egg yolk of Chinese soft-shelled turtle.

Key words: Chinese soft-shelled turtle egg yolk; angiotensin converting enzyme inhibitory peptide; isolation and purification; bioinformatics; molecular docking

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