辣木籽ACE抑制肽的分离纯化、结构鉴定及其体外活性评价  

Isolation,Structural Identification and in Vitro Activity Evaluation of Angiotensin-Converting Enzyme Inhibitory Peptides from Moringa oleifera Seeds

作　　者：钟玉旺 徐万莉 范尧珠 黎依艳 王雪峰[1] ZHONG Yuwang;XU Wani;FAN Yaozhu;LI Yiyan;WANG Xuefeng(College of Food Science and Technology,Yunnan Agricultural University,Kunming 650201,China;Lijiang Vocational and Technical College,Lijiang 674100,China;Ziyang Vocational College of Environmental Science and Technology,Ziyang 641300,China)

机构地区：[1]云南农业大学食品科学技术学院,云南昆明650201 [2]丽江职业技术学院,云南丽江674100 [3]资阳环境科技职业学院,四川资阳641300

出　　处：《食品科学》2023年第24期118-126,共9页Food Science

基　　金：国家自然科学基金地区科学基金项目(31960462);云南省基础研究计划面上项目(2019FB052);云南省农业联合专项重点项目(202101BD070001-013);云南省科学技术协会青年科技人才托举工程项目。

摘　　要：采用超滤、离子交换层析分离辣木籽蛋白酶解产物,以血管紧张素转换酶(angiotensin-converting enzyme,ACE)抑制率为评价指标,筛选具有较高降压活性的肽组分,并通过高效液相色谱-串联质谱鉴定其肽序列,结合生物信息学和分子对接技术筛选出潜在的ACE抑制肽,进一步利用傅里叶变换红外光谱、酶反应抑制剂动力学和四甲基偶氮唑蓝法实验解析其二级结构特征及体外活性。结果表明:分离得到的F-b肽组分具有较好的降压效果,共鉴定出11条肽序列,进一步筛选出肽QGPRPQ为潜在的ACE抑制肽(IC_(50)=(1.15±0.3)mmol/L),分子对接表明QGPRPQ可以与ACE以氢键、疏水作用更好地结合;二级结构分析表明QGPRPQ由22.8%α-螺旋、33.3%β-折叠和43.9%β-转角构成;QGPRPQ的抑制模型为混合型抑制,且质量浓度低于0.01 mg/mL时对HepG2细胞无毒性作用。该研究可为辣木籽蛋白源降压肽的开发利用提供一定的理论参考。In this study,angiotensin-converting enzyme(ACE)inhibitory peptides from an enzymatic hydrolysate of Moringa oleifera seeds were separated by sequential ultrafiltration and ion exchange chromatography.The peptide sequences were identified by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)and potential ACE inhibitory peptides were selected by bioinformatics and molecular docking;their secondary structure was analyzed by Fourier transform infrared(FTIR)spectroscopy and their in vitro activity was evaluated by enzymatic inhibition kinetics and the methyl thiazolyl tetrazolium(MTT)method.The results showed that peptide fraction F-b had a good antihypertensive effect.A total of 11 peptide sequences were identified.Peptide QGPRPQ was identified as a potential ACE inhibitory peptide with a half-maximum inhibitory concentration(IC_(50))(1.15±0.3)mmol/L.Molecular docking showed that QGPRPQ could better bind to ACE through hydrogen bond and hydrophobic interaction.Secondary structure analysis showed that QGPRPQ was composed of 22.8%α-helix,33.3%β-fold and 43.9%β-turn.The mode of inhibition of QGPRPQ was mixed type,and it had no toxic effect on HepG2 cells at a concentration lower than 0.01 mg/mL.This study can provide an important theoretical basis for the development and utilization of hypotensive peptides derived from M.oleifera seed protein.

关 键 词：辣木籽 ACE抑制肽 高效液相色谱-串联质谱 分子对接 二级结构 酶反应抑制动力学 

分 类 号：TS201.2[轻工技术与工程—食品科学]