杂色蛤中ACE抑制肽的分离鉴定与分子对接研究

Separation and identification of ACE inhibitory peptides from Ruditapes philippinarum and molecular docking

目的:建立一种快速、准确筛选杂色蛤酶解物中活性多肽的方法。方法:在ACE抑制活性导向下,以超滤技术与离子交换法对杂色蛤酶解物进行分离,确定ACE抑制活性多肽,通过质谱分析与已有序列比对的方法,确定活性多肽的氨基酸序列;通过计算机模拟将多肽与ACE蛋白对接,筛选活性多肽,并确定其与ACE蛋白的作用位点。结果:共鉴定67个多肽,其中QIIVQDLTKR、LDYRPGDKFKGT、NTQIIVQDLTKR和LLFDRAPVNFGNYR这四个多肽能与ACE稳定结合,ACE的Glu403为一重要的结合位点,且Ala356和Arg522对多肽与ACE的稳定结合也产生了重要影响。结论:基于串联质谱与分子对接技术,建立从混合多肽中快速筛选、鉴定活性多肽的方法,明确活性多肽与ACE可能的结合位点,为后续的深入研究提供了依据和参考。

Objective: To select the bioactive peptides quickly and accurately from Ruditapes philippinarum hydrolysate.Methods: Peptides from Ruditapes philippinarum hydrolysate could be separated by ultrafiltration and ion exchange chromatography based on the ACE inhibitory assay- direct fractionation. Then amino acids sequences of peptides were identified according to the MS / MS spectra and protein database. Interaction mechanism between the peptides and ACE was investigated by molecular docking. Results: 67 peptides were identified,among which four peptides,QIIVQDLTKR,LDYRPGDKFKGT,NTQIIVQDLTKR and LLFDRAPVNFGNYR,can bind to ACE stably. Our docking results also suggested that the ACE inhibitory peptides bind to ACE via interactions with Glu403,Ala356 and Arg522 residues and the residue Glu403 might be a significant binding site.Conclusions: this study can quickly select,identify and evaluate the bioactive peptides while finding potential binding sites between bioactive peptides and ACE,which provides the basis and reference for further research.