| 聂鹏,孟乐,李针,等.单环刺螠ACE抑制肽的分离、鉴定及活性评价[J].中国海洋药物,2024,43(3):51-59. |
| 单环刺螠ACE抑制肽的分离、鉴定及活性评价 |
| Isolation, identification and activity evaluation of ACE inhibitory peptides from Urechis unicinctus |
| 投稿时间：2023-03-02 修订日期：2023-04-07 |
| DOI：10.13400/j.cnki.cjmd.2024.03.010 |
| 中文关键词: 单环刺螠 酶解肽 ACE抑制活性 分离纯化 |
| English Keywords:Urechis Unicinctus hydrolyzed peptides ACE inhibitory activity isolation and purification |
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| 中文摘要: |
| 目的：以单环刺螠(Urechis unicinctus)为原材料,以血管紧张素转化酶(ACE)抑制活性为筛选指标,运用酶解技术和现代分离纯化技术,制备具有降压潜能的ACE抑制肽。方法：运用可控酶解技术,以ACE的抑制率为筛选指标,优化酶解条件。运用超滤、Sephadex G-15凝胶柱层析、反相高效液相色谱(RP-HPLC)及质谱测序等一系列分离纯化和鉴定技术,制备具有ACE抑制活性的生物活性肽,并通过Lineweaver-Burk法确定其抑制剂类型。结果：最佳酶解条件为：胰蛋白酶,37℃、pH 8.0、加酶量4.0%、料液比1:30、酶解时间4.0 h。通过超滤、Sephadex G-15凝胶层析和反相高效液相色谱(RP-HPLC)进行分离纯化,得到5条活性肽,经质谱和氨基酸序列分析确定其分别为：Pro-Gln-Met-Thr-Phe(DHCY1, 622.75 Da)、Pro-Tyr-Phe-Lys-His(DHCY2, 690.8 Da)、Pro-Gly-Trp-Lys-Ala(DHCY3, 557.66 Da)、Pro-Gln-Gly-Met-Ile-Val-Val(DHCY4, 742.94 Da)、Val-Met-Arg-Ile-Ile(DHCY5, 630.86 Da)。其中DHCY1、DHCY4、DHCY5表现出较好的ACE抑制活性,其抑制率分别为86.26±0.85%、92.11±1.13%、96.51±1.04%。经判断,DHCY5为竞争性ACE抑制剂。结论：胰蛋白酶酶解单环刺螠蛋白,可以产生具有显著ACE抑制活性的酶解肽,包括ACE竞争性抑制剂,具有潜在的降血压活性,可为单环刺螠酶解肽的开发利用和降血压产品的开发提供理论依据和参考。 |
| English Summary: |
| Objective: Urechis unicinctus was used as raw material to prepare ACE inhibitory peptides with hypotensive potential by enzymatic hydrolysis and modern separation and purification techniques. ACE inhibitory activity was used as screening index. Methods: The inhibition rate of ACE was used as the screening index, controlled enzymatic hydrolysis technology was used to optimize the conditions of enzymatic hydrolysis. A series of separation, purification and identification techniques including ultrafiltration, Sephadex G-15 gel column chromatography, reversed phase high performance liquid chromatography (RP-HPLC) and mass spectrometry sequencing were used to prepare bioactive peptides with ACE inhibitory activity, the inhibitor types were determined by Lineweaver-Burk method. Results: The optimum conditions were trypsin, 37 ℃, pH 8.0, 4.0% enzyme dosage, 1:30 feed-liquid ratio and 4.0 h enzymolysis time. Five active peptides were isolated, purified and identified as Pro-Gln-Met-Thr-Phe (DHCY1, 622.75 Da), Pro-Tyr-Phe-Lys-His (DHCY2, 690.8 Da), Pro-Gly-Trp-Lys-Ala (DHCY3, 557.66 Da), Pro-Gln-Gly-Met-Ile-Val-Val (DHCY4, 742.94 Da) and Val-Met-Arg-Ile-Ile (DHCY5, 630.86 Da) by mass spectrometry and amino acid sequence analysis. Among them, DHCY1, DHCY4 and DHCY5 showed better ACE inhibitory activity, the inhibitory rates were 86.26 ± 0.85%, 92.11±1.13% and 96.51±1.04%, respectively. DHCY5 were judged to be a competitive ACE inhibitor. Conclusion: The trypsin hydrolyzes Urechis Unicinctus proteins can produce peptides with significant ACE inhibitory activity, including ACE competitive inhibition, which have potential antihypertensive activity, it can provide the theoretical basis and reference for the exploitation and utilization of U. Unicinctus enzymatic peptides and the development of antihypertensive products. |
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