基于生物信息学与分子对接技术对坛紫菜降血压肽的筛选及活性分析

doi:10.7506/spkx1002-6630-20201021-201

摘要/Abstract

摘要： 采用木瓜蛋白酶和糜蛋白酶双酶联合水解坛紫菜蛋白（Porphyra haitanensis protein，PHP）制备降血压活性肽，测定了经超滤后各个分子质量PHP酶解液清除1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl，DPPH）自由基和2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)（2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)，ABTS）阳离子自由基能力、铁离子还原抗氧化能力（ferric ion reducing antioxidant power，FRAP）以及血管紧张素转化酶（angiotensin converting enzyme，ACE）抑制活性。经质谱鉴定和活性肽数据库BIOPEP以及AHTPDB网站查找筛选活性肽，采用分子对接解释多肽抑制ACE机理，并对双酶酶解PHP动力学进行研究。结果表明：经超滤后的8?个分子质量的坛紫菜多肽组分中，＜0.5?kDa的组分具有最高DPPH自由基和ABTS阳离子自由基清除能力，＜1?kDa的组分具有最高FRAP值，而0.5～1?kDa组分IC50值（（2.21±0.28）mg/mL）最低。经查找筛选出14?条降血压肽，其中4?条肽的序列和对接能量分别为AVP（－5.87?kJ/mol）、GPL（－6.13?kJ/mol）、LDY（－7.65?kJ/mol）和LGYL（－7.78?kJ/mol）。酶解动力学模型预测PHP水解度与实验水解度相对误差在10%以下。本实验基于串联质谱与分子对接技术，通过生物信息筛选和体外活性检测从混合多肽中快速鉴定、筛选活性多肽，探究多肽抑制ACE的机理，并建立了描述蛋白可控酶解过程规律的动力学模型，为活性肽的制备提供了参考和借鉴。

关键词: 坛紫菜；血管紧张素转化酶抑制肽；分子对接；酶解动力学模型

Abstract: Porphyra haitanensis protein (PHP) was simultaneously hydrolyzed with papain and chymotrypsin to prepare antihypertensive peptides. The hydrolyate was fractionated by ultrafiltration, and the resulting fractions were evaluated for scavenging capacity toward 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) cation radicals, ferric ion reducing antioxidant power (FRAP) and angiotensin converting enzyme (ACE) inhibitory activity. The bioactive peptides were identified by mass spectrometry (MS) as well as through search in the bioactive peptides (BIOPEP) and antihypertensive peptides (AHTPDB) databases, and their ACE inhibitory mechanism was elucidated by molecular docking. Additionally, the kinetics of enzymatic hydrolysis of PHP was studied. The results showed that among the eight fractions obtained after ultrafiltration, the fraction with molecular mass less than 0.5 kDa had the highest DPPH and ABTS cation radical scavenging capacity, the fraction with molecular mass less than 1 kDa had the highest FRAP value, and the fraction with molecular mass in the range of 500–1 000 Da had the lowest half-maximal inhibitory concentration (IC50) value ((2.21 ± 0.28) mg/mL). Totally, 14 bioactive peptides were identified, four of which were selected for molecular docking analysis, indicating that their amino acid sequences were and docking energy were AVP (?5.87 kJ/mol), GPL (?6.13 kJ/mol), LDY (?7.65 kJ/mol) and LGYL (?7.78 kJ/mol), respectively. The relative error between the hydrolysis degree of PHP predicted by the kinetic model of enzymatic hydrolysis and the experimental values was less than 10%. This study can provide reference for the preparation of bioactive peptides.

Key words: Porphyra haitanensis; angiotensin converting enzyme inhibitory peptides; molecular docking; enzymatic hydrolysis kinetics

中图分类号:

TS209

叶贤江，苏志琛，林晓娟，陈继承. 基于生物信息学与分子对接技术对坛紫菜降血压肽的筛选及活性分析[J]. 食品科学, 2021, 42(24): 140-148.

YE Xianjiang, SU Zhichen, LIN Xiaojuan, CHEN Jicheng. Screening and Activity Evaluation of Antihypertensive Peptides from Porphyra haitanensis by Bioinformatics and Molecular Docking[J]. FOOD SCIENCE, 2021, 42(24): 140-148.

[1]	叶贤江苏志琛林晓娟陈继承. 基于生物信息学与分子对接技术对坛紫菜降血压肽的筛选及活性分析[J]. 食品科学, 0, (): 0-0.
[2]	侯成杰聂彩清王彦茜艾连中夏永军张汇谢凡王光强. α-乳白蛋白源ACE抑制肽精准筛选及验证[J]. , 0, (): 0-0.
[3]	许馨予杨鹄隽贾斌张慧敏左锋. 大豆分离蛋白-高酯柑橘果胶-没食子酸复合Pickering乳液制备及其稳定性分析[J]. 食品科学, 0, (): 0-0.
[4]	杨舒乔高彦祥王迪. 非变性II型胶原蛋白研究进展[J]. , 0, (): 0-0.
[5]	梁曾恩妮刘伟汪秋安张菊华苏东林付复华李高阳单杨. 圣草次苷的合成及其对过氧化氢诱导HUVEC细胞氧化损伤的保护作用[J]. , 0, (): 0-0.
[6]	陈永凯张鸿儒刘泓张春晖姜珊郭玉杰. 促成骨细胞增殖活性骨胶原蛋白肽的靶向筛选及活性分析[J]. 食品科学, 0, (): 0-0.
[7]	刘静波，王子秦，于一丁，张婷，刘博群. 豆粕血管紧张素转化酶抑制肽的结构鉴定及作用机制解析[J]. 食品科学, 2021, 42(12): 123-129.
[8]	刘静波王子秦于一丁张婷刘博群. 豆粕血管紧张素转化酶抑制肽的结构鉴定及作用机制解析[J]. 食品科学, 0, (): 0-0.
[9]	方东路，马晓惠，赵明文，郑惠华，陈惠，胡秋辉，赵立艳. 添加灰树花粉面团的酶法改性及面条品质评价[J]. 食品科学, 2021, 42(10): 23-31.
[10]	阴宏婕鞠化鹏钟利敏孙娜林松毅. 核黄素结合肽的生物活性及结构表征[J]. , 0, (): 0-0.
[11]	阴宏婕，鞠化鹏，钟利敏，孙娜，林松毅. 核黄素结合肽的生物活性及结构表征[J]. 食品科学, 2021, 42(9): 137-144.
[12]	张欢戴宏杰陈媛马良张宇昊. 离子液体-球磨法制备柠檬籽纤维素纳米纤丝及其结构表征[J]. , 0, (): 0-0.
[13]	张欢，戴宏杰，陈媛，高焕秋，刘姣，马良，余永，张宇昊. 离子液体-球磨法制备柠檬籽纤维素纳米纤丝及其结构表征[J]. 食品科学, 2021, 42(7): 120-127.
[14]	陈慧, YOSHINORI Katakura, 扈洪波, 尹淑涛, 赵冲. SIRT3多酚激活剂的筛选及其对UVB诱导HaCaT细胞衰老的修复作用[J]. 食品科学, 2021, 42(5): 115-121.
[15]	陈慧 Katakura Yoshinori 扈洪波尹淑涛赵冲. SIRT3多酚激活剂的筛选及其对UVB诱导皮肤细胞衰老的修复作用[J]. 食品科学, 0, (): 0-0.

基于生物信息学与分子对接技术对坛紫菜降血压肽的筛选及活性分析

Screening and Activity Evaluation of Antihypertensive Peptides from Porphyra haitanensis by Bioinformatics and Molecular Docking

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价