利用牡蛎制备DPP-IV抑制肽及其活性分析

doi:10.7506/spkx1002-6630-20200226-294

食品科学 ›› 2021, Vol. 42 ›› Issue (10): 120-126.doi: 10.7506/spkx1002-6630-20200226-294

利用牡蛎制备DPP-IV抑制肽及其活性分析

陈宏，章骞，陈玉磊，翁凌，张凌晶，谢少浩，刘光明，曹敏杰

（1.集美大学食品与生物工程学院，福建厦门 361021；2.大连工业大学海洋食品深加工协同创新中心，辽宁大连 116034）

出版日期:2021-05-25 发布日期:2021-06-02
基金资助:
“十三五”国家重点研发计划重点专项（2018YFD0901004）；财政部和农业农村部：国家现代农业产业技术体系资助项目（CARS-49）

Preparation and Activity Analysis of DPP-IV Inhibitory Peptides from Pacific Oyster (Crassostrea gigas)

CHEN Hong, ZHANG Qian, CHEN Yulei, WENG Ling, ZHANG Lingjing, XIE Shaohao, LIU Guangming, CAO Minjie

(1. College of Food and Biological Engineering, Jimei University, Xiamen 361021, China;2. Collaborative Innovation Center of Marine Food Deep Processing, Dalian Polytechnic University, Dalian 116034, China)

Online:2021-05-25 Published:2021-06-02

摘要/Abstract

摘要： 以牡蛎肉为原料制备二肽基肽酶-IV（dipeptidyl peptidase-IV，DPP-IV，EC 3.4.14.5）抑制肽。通过对比5 种蛋白酶（木瓜蛋白酶、碱性蛋白酶、胰酶、中性蛋白酶和复合蛋白酶）制备的酶解液对DPP-IV的抑制活性，发现胰酶制备的酶解液对DPP-IV抑制效果最好。通过优化胰酶酶解条件，确定最优酶解条件为加酶量0.8%、pH 8.0、温度37 ℃，酶解时间90 min。酶解液经超滤、Sephadex G-15和高效液相色谱分离纯化，获得肽片段。通过质谱鉴定筛选得到2 种肽段的序列为Glu-Ile-Thr-Ala-Leu-Ala-Pro-Ser-Thr-Met-Lys（EITALAPSTMK）和Ile-Leu-Ala-Pro-Pro-Glu-Arg（ILAPPER）。利用BIOPEP在线模拟分析了这2 个肽的胃肠液消化特性。采用固相合成法合成肽段APSTM和ILAPPER，并应用于DPP-IV抑制活性研究。结果显示，2 个肽段的IC50值分别为354.81 μmol/L和16.98 μmol/L。分子对接结果表明，抑制肽与DPP-IV的活性部位主要以氢键、范德华力和π键相互作用为主。本研究通过酶解牡蛎肉制备高抑制活性的DPP-IV抑制肽，为以牡蛎为原料的功能性食品开发利用提供了理论基础。

关键词: 牡蛎；二肽基肽酶-IV抑制肽；分离纯化；抑制活性；分子对接

Abstract: To prepare dipeptidyl peptidase-IV (DPP-IV) inhibitory peptides, Pacific oyster flesh (Crassostrea gigas) was hydrolyzed with five different proteases, including papain, alcalase, pancreatin, neutrase and protromex. The pancreatin hydrolysate was found to have higher DPP-IV inhibitory activity as compared with the other four hydrolysates. The optimal hydrolysis conditions with pancreatin were determined to be 0.8%, 8.0, 37 ℃ and 90 min for enzyme dosage, pH, temperature, and hydrolysis duration, respectively. The hydrolysate obtained using the optimized conditions was purified and fractionated sequentially by ultrafiltration, Sephadex G-15 column chromatography and high performance liquid chromatography (HPLC) into different peptide fractions. Two DPP-IV inhibitory peptides were obtained and identified as Glu-Ile-Thr-Ala-Leu-Ala-Pro-Ser-Thr-Met-Lys (EITALAPSTMK) and Ile-Leu-Ala-Pro-Pro-Glu-Arg (ILAPPER) by mass spectrometry (MS). Their digestibility characteristics in gastrointestinal fluid were analyzed using online simulation with BIOPEP. Two peptides APSTM and ILAPPER were synthesized by solid-phase synthesis for DPP-IV inhibitory activity evaluation. The results showed that the half-maximal inhibitory concentration (IC50) values of the two peptides were 354.81 and 16.98 mmol/L, respectively. Molecular docking results indicated that inhibitory peptides interacted with the active site of DPP-IV through hydrogen bonds, van der Waals forces and p bonds. The results of this study provide a theoretical basis for the development of functional foods from oyster in the future.

Key words: oyster; dipeptidyl peptidase-IV inhibitory peptide; isolation and purification; inhibitory activity; molecular docking

中图分类号:

TS254.1

陈宏，章骞，陈玉磊，翁凌，张凌晶，谢少浩，刘光明，曹敏杰. 利用牡蛎制备DPP-IV抑制肽及其活性分析[J]. 食品科学, 2021, 42(10): 120-126.

CHEN Hong, ZHANG Qian, CHEN Yulei, WENG Ling, ZHANG Lingjing, XIE Shaohao, LIU Guangming, CAO Minjie. Preparation and Activity Analysis of DPP-IV Inhibitory Peptides from Pacific Oyster (Crassostrea gigas)[J]. FOOD SCIENCE, 2021, 42(10): 120-126.

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利用牡蛎制备DPP-IV抑制肽及其活性分析

Preparation and Activity Analysis of DPP-IV Inhibitory Peptides from Pacific Oyster (Crassostrea gigas)

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