珍珠蚌肉二肽基肽酶-Ⅳ抑制肽的制备与分离

TS254.9; 目的:以珍珠蚌肉为原料制备对二肽基肽酶(dipeptidyl peptidase,DPP)-Ⅳ具有抑制作用的生物活性肽.方法:利用两步酶解法并优化珍珠蚌肉酶解条件,筛选体外DPP-Ⅳ抑制活性较高组分进行超滤膜浓缩,SuperdexTM peptide 10/300 GL凝胶柱层析与反相高效液相色谱(reverse phase-high performance liquid chromatography,RP-HPLC)技术分离纯化出酶解液中的DPP-Ⅳ抑制肽,最终通过液相色谱-串联质谱鉴定其氨基酸序列.结果:中性蛋白酶与碱性蛋白酶对珍珠蚌肉均有良好降解效果,经3 kDa超滤膜...

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Published in食品科学 Vol. 45; no. 17; pp. 63 - 70
Main Authors 张楚涵, 陈寅格, 马婷, 翁凌, 张凌晶, 张根芳, 曹敏杰
Format Magazine Article
LanguageChinese
Published 集美大学体育学院,福建厦门 361021 15.09.2024
集美大学海洋食品与生物工程学院,福建厦门 361021%集美大学海洋食品与生物工程学院,福建厦门 361021%金华职业技术大学农学院,浙江金华 321000
Subjects
separation and purification
抑制肽
pearl mussel
珍珠蚌
inhibitory peptides
二肽基肽酶-
dipeptidyl peptidase-
分离纯化
enzymatic hydrolysis
酶解
Online AccessGet full text
ISSN1002-6630
DOI10.7506/spkx1002-6630-20240125-233

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Abstract TS254.9; 目的:以珍珠蚌肉为原料制备对二肽基肽酶(dipeptidyl peptidase,DPP)-Ⅳ具有抑制作用的生物活性肽.方法:利用两步酶解法并优化珍珠蚌肉酶解条件,筛选体外DPP-Ⅳ抑制活性较高组分进行超滤膜浓缩,SuperdexTM peptide 10/300 GL凝胶柱层析与反相高效液相色谱(reverse phase-high performance liquid chromatography,RP-HPLC)技术分离纯化出酶解液中的DPP-Ⅳ抑制肽,最终通过液相色谱-串联质谱鉴定其氨基酸序列.结果:中性蛋白酶与碱性蛋白酶对珍珠蚌肉均有良好降解效果,经3 kDa超滤膜浓缩后,酶解产物的DPP-Ⅳ抑制率可达67.4％.通过凝胶过滤柱和RP-HPLC进一步分离纯化出6个活性较高的肽段,序列分别为FNAPAM、FIPNY、IYNPPTPF、LAMPYP、FFVVMP和LAGMP.进一步,以分子对接的方式分析6个肽段与DPP-Ⅳ的相互作用关系.结论:本研究利用珍珠蚌肉制备DPP-Ⅳ抑制肽,可为以水产加工副产物为原料制备功能性食品提供理论参考.
AbstractList TS254.9; 目的:以珍珠蚌肉为原料制备对二肽基肽酶(dipeptidyl peptidase,DPP)-Ⅳ具有抑制作用的生物活性肽.方法:利用两步酶解法并优化珍珠蚌肉酶解条件,筛选体外DPP-Ⅳ抑制活性较高组分进行超滤膜浓缩,SuperdexTM peptide 10/300 GL凝胶柱层析与反相高效液相色谱(reverse phase-high performance liquid chromatography,RP-HPLC)技术分离纯化出酶解液中的DPP-Ⅳ抑制肽,最终通过液相色谱-串联质谱鉴定其氨基酸序列.结果:中性蛋白酶与碱性蛋白酶对珍珠蚌肉均有良好降解效果,经3 kDa超滤膜浓缩后,酶解产物的DPP-Ⅳ抑制率可达67.4％.通过凝胶过滤柱和RP-HPLC进一步分离纯化出6个活性较高的肽段,序列分别为FNAPAM、FIPNY、IYNPPTPF、LAMPYP、FFVVMP和LAGMP.进一步,以分子对接的方式分析6个肽段与DPP-Ⅳ的相互作用关系.结论:本研究利用珍珠蚌肉制备DPP-Ⅳ抑制肽,可为以水产加工副产物为原料制备功能性食品提供理论参考.
Abstract_FL Objective:Bioactive peptides with an inhibitory effect on dipeptidyl peptidase-Ⅳ(DPP-Ⅳ)were prepared from pearl mussel muscle.Methods:Two-step enzymatic hydrolysis was adopted and enzymatic hydrolysis conditions were optimized.Fractions with in vitro DPP-Ⅳ inhibitory activity were concentrated by ultrafiltration,and purified by SuperdexTM peptide 10/300 GL gel filtration column chromatography and reverse phase-high performance liquid chromatography(RP-HPLC).DPP-Ⅳ inhibitory peptides were purified from the enzymatic hydrolysate and their amino acid sequences were identified by liquid chromatography-tandem mass spectrometry.Results:Both neutral protease and alkaline protease could effectively hydrolyze pearl mussel muscle.After concentration using a 3 kDa cut-off ultrafiltration membrane,the DPP-Ⅳ inhibition rate of the enzymatic hydrolysate reached 67.4％.After separation by gel filtration column and purification by RP-HPLC,6 active peptides were obtained,whose sequences were FNAPAM,FIPNY,IYNPPTPF,LAMPYP,FFVVMP and LAGMP,respectively.Furthermore,the interactions between these peptides and DPP-Ⅳ were analyzed by molecular docking.Conclusion:Our present study provides a theoretical reference for the effective utilization of fish processing byproducts as raw materials for functional food production.
Author 陈寅格
翁凌
张凌晶
曹敏杰
马婷
张根芳
张楚涵
AuthorAffiliation 集美大学体育学院,福建厦门 361021;集美大学海洋食品与生物工程学院,福建厦门 361021%集美大学海洋食品与生物工程学院,福建厦门 361021%金华职业技术大学农学院,浙江金华 321000
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Author_FL ZHANG Lingjing
WENG Ling
ZHANG Genfang
CHEN Yinge
ZHANG Chuhan
CAO Minjie
MA Ting
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Copyright Copyright © Wanfang Data Co. Ltd. All Rights Reserved.
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DocumentTitle_FL Preparation and Purification of Dipeptidyl Peptidase-Ⅳ Inhibitory Peptides from Pearl Mussel Muscle
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Issue 17
Keywords separation and purification
抑制肽
pearl mussel
珍珠蚌
inhibitory peptides
二肽基肽酶-
dipeptidyl peptidase-
分离纯化
enzymatic hydrolysis
酶解
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PublicationTitle 食品科学
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Publisher 集美大学体育学院,福建厦门 361021
集美大学海洋食品与生物工程学院,福建厦门 361021%集美大学海洋食品与生物工程学院,福建厦门 361021%金华职业技术大学农学院,浙江金华 321000
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Snippet TS254.9; 目的:以珍珠蚌肉为原料制备对二肽基肽酶(dipeptidyl peptidase,DPP)-Ⅳ具有抑制作用的生物活性肽.方法:利用两步酶解法并优化珍珠蚌肉酶解条件,筛选体外DPP-Ⅳ抑制...
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StartPage 63
Title 珍珠蚌肉二肽基肽酶-Ⅳ抑制肽的制备与分离
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