马乳清蛋白ACE、DPP-Ⅳ、AG抑制肽的分离及鉴定

doi:10.16431/j.cnki.1671-7236.2025.12.011

中国畜牧兽医 ›› 2025, Vol. 52 ›› Issue (12): 5651-5660.doi: 10.16431/j.cnki.1671-7236.2025.12.011

• 营养与饲料 • 上一篇

马乳清蛋白ACE、DPP-Ⅳ、AG抑制肽的分离及鉴定

娄肖肖^1,2, 赵艳坤^2,3, 郑楠³, 陈贺², 马梅², 邵伟¹

1. 新疆农业大学动物科学学院, 新疆奶业工程技术研究中心, 乌鲁木齐 830063;
2. 新疆维吾尔自治区农业科学院农业质量标准与检测技术研究所, 农业农村部农产品质量安全风险评估实验室, 新疆农产品质量安全实验室, 乌鲁木齐 830091;
3. 中国农业科学院北京畜牧兽医研究所, 农业农村部奶及奶制品质量安全控制重点实验室, 北京 100193

修回日期:2025-08-01 发布日期:2025-11-28
通讯作者: 邵伟 E-mail:dksw@xjau.edu.cn
作者简介:娄肖肖，E-mail,Louxiaoxiao1024@163.com。
基金资助:
新疆维吾尔自治区重点研发项目(2023B02034-1)；国家重点研发计划(2022YFD1600103)；新疆维吾尔自治区重大科技专项(2022A02006-3-2)；新疆维吾尔自治区现代农业产业技术体系(XJARS-11-09)

Isolation and Identification of ACE,DPP-Ⅳ,and AG Inhibitory Peptides from Mare Whey Protein

LOU Xiaoxiao^1,2, ZHAO Yankun^2,3, ZHENG Nan³, CHEN He², MA Mei², SHAO Wei¹

1. Xinjiang Dairy Engineering Technology Research Center, College of Animal Science, Xinjiang Agricultural University, Urumqi 830063, China;
2. Xinjiang Agricultural Product Quality and Safety Risk Assessment Laboratory, Laboratory of Quality and Safety Risk Assessment for Agro-Products, Ministry of Agriculture and Rural Affairs, Institute of Quality Standards & Testing Technology for Agro-Products, Xinjiang Uyghur Autonomous Region Academy of Agricultural Sciences, Urumqi 830091, China;
3. Key Laboratory for Quality & Safety Control for Milk and Dairy Products of Ministry of Agriculture and Rural Affairs, Institute of Animal Science of CAAS, Beijing 100193, China

Revised:2025-08-01 Published:2025-11-28

摘要/Abstract

摘要： 【目的】从马乳清蛋白水解产物中分离鉴定对血管紧张素转换酶(ACE)、二肽基肽酶-Ⅳ(DPP-Ⅳ)和α-葡萄糖苷酶(AG)具有多重抑制活性的多肽。【方法】自新疆乌鲁木齐南山牧场采集15份生马乳样品，以马乳清蛋白为底物，分别用单酶(胰蛋白酶、中性蛋白酶和碱性蛋白酶)、双酶(胰蛋白酶＋中性蛋白酶)和三酶(胰蛋白酶＋中性蛋白酶＋碱性蛋白酶)进行水解，通过超滤法对水解产物进行分离；通过测定ACE抑制活性评价各级分离产物的降血压活性，通过测定AG抑制活性和DPP-Ⅳ抑制活性评价各级分离产物的降血糖活性；通过液质联用(LC-MS/MS)的方法鉴定多肽的氨基酸组成，根据鉴定结果化学合成活性较好的多肽，并体外测定其抑制活性。【结果】与单酶、双酶水解产物相比，三酶水解产物的水解度(74.33%)、ACE抑制活性(22.81%)、DPP-Ⅳ抑制活性(23.46%)及AG抑制活性(20.77%)均显著提高(P＜0.05)。从活性最优的三酶水解产物中分离鉴定出具有多重抑制活性的新型多肽系列(HSVAMAASDISLLDSESAPLR、TQMVDEEIMEK、VQIVPDLTR、RALQPLPGRVQIVPDLTR、TELTEEEKNYLK、LRPTPEDNLEIILR)；体外化学合成这几种多肽，并测定其活性，结果显示，TQMVDEEIMEK对ACE抑制率达99.89%，VQIVPDLTR对DPP-Ⅳ抑制率达90.85%，TELTEEEKNYLK对AG抑制率达97.87%。【结论】本研究从马乳清蛋白水解产物中成功分离鉴定出6种具有ACE、DPP-Ⅳ、AG多重抑制活性的新型多肽，为马乳功能性产品的开发提供了理论依据。

关键词: 马乳清蛋白; 生物活性肽; 抑制活性

Abstract: 【Objective】 This study aimed to isolate and identify peptides with multiple inhibitory activities against angiotensin-converting enzyme (ACE),dipeptidyl peptidase-Ⅳ (DPP-Ⅳ),and α-glucosidase (AG) from hydrolyzed mare whey protein. 【Method】 Fifteen raw mare milk samples were collected from Nanshan Pasture in Urumqi,Xinjiang.Using mare whey protein as the substrate,hydrolysis was performed with single enzymes (trypsin,neutral protease,and alkaline protease),a two-enzyme combination (trypsin + neutral protease),and a three-enzyme combination (trypsin＋neutral protease＋alkaline protease).The hydrolyzed products were separated by ultrafiltration.The antihypertensive activity of the separated products was evaluated by determining ACE inhibitory activity,while their antidiabetic activity was assessed by measuring AG inhibitory activity and DPP-Ⅳ inhibitory activity.The amino acid composition of the peptides was identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS).Based on the identification results,peptides with better activity were chemically synthesized,and their inhibitory activity was determined in vitro. 【Result】 Compared with the products of single-enzyme and two-enzyme hydrolysis,the three-enzyme hydrolyzates showed significantly higher performance (P<0.05),with a hydrolysis degree of 74.33%,ACE inhibitory activity of 22.81%,DPP-Ⅳ inhibitory activity of 23.46%,and AG inhibitory activity of 20.77%.From the three-enzyme hydrolyzates with the optimal activity,a series of novel peptides with multiple inhibitory activities were isolated and identified,including HSVAMAASDISLLDSESAPLR,TQMVDEEIMEK,VQIVPDLTR,RALQPLPGRVQIVPDLTR,TELTEEEKNYLK,and LRPTPEDNLEIILR.These peptides were chemically synthesized in vitro,and their activities were determined.The results showed that TQMVDEEIMEK exhibited an ACE inhibition rate of 99.89%,VQIVPDLTR had a DPP-Ⅳ inhibition rate of 90.85%,and TELTEEEKNYLK showed an AG inhibition rate of 97.87%. 【Conclusion】 In this study, six novel peptides with multiple inhibitory activities against ACE,DPP-Ⅳ,and AG from mare whey protein hydrolyzates were successfully isolated and identified,laying a theoretical foundation for the development of functional mare milk products.

Key words: mare whey protein; bioactive peptides; inhibitory activity

中图分类号:

S82
TS252.1

娄肖肖, 赵艳坤, 郑楠, 陈贺, 马梅, 邵伟. 马乳清蛋白ACE、DPP-Ⅳ、AG抑制肽的分离及鉴定[J]. 中国畜牧兽医, 2025, 52(12): 5651-5660.

LOU Xiaoxiao, ZHAO Yankun, ZHENG Nan, CHEN He, MA Mei, SHAO Wei. Isolation and Identification of ACE,DPP-Ⅳ,and AG Inhibitory Peptides from Mare Whey Protein[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(12): 5651-5660.

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马乳清蛋白ACE、DPP-Ⅳ、AG抑制肽的分离及鉴定

Isolation and Identification of ACE,DPP-Ⅳ,and AG Inhibitory Peptides from Mare Whey Protein

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