文献类型：期刊文献

中文题名：酪蛋白抗凝血肽的制备及其体外抗凝血、溶栓作用研究

英文题名：Preparation of anticoagulant peptide from casein and study on thrombolysis and anticoagulation in vitro

作者：苗艳丽[1];刘卓勤[1];孙瑞雄[1];李卫东[1];廖艳[2];陈绍红[2]

机构：[1]广东海洋大学理学院,广东湛江524088;[2]广东海洋大学现代生物化学中心,广东湛江524088

年份：2016

卷号：47

期号：11

起止页码：1875

中文期刊名：中草药

外文期刊名：Chinese Traditional and Herbal Drugs

收录：CSTPCD、、北大核心2014、CSCD2015_2016、Scopus、北大核心、CSCD

基金：广东省湛江市科技计划项目(2013B01073);广西科学研究与计划开发项目(桂科合14125008-1-10);广东海洋大学创新强校项目(GDOU2015050209);广东省教育厅2015年重点平台及科研项目(2015KTSCX057)

语种：中文

中文关键词：抗凝血肽;酪蛋白;抗凝血;溶栓;木瓜蛋白酶;菠萝蛋白酶;中性蛋白酶;碱性蛋白酶;凝血酶;高效体积排阻色谱;马尿酰-组氨酰-亮氨酸

外文关键词：anticoagulant peptide; casein; anticoagulant activity; thrombolytic; papaya protease; pineapple protease; neutral protease; alkaline protease; thrombin; high performance size exclusion chromatography; N-Hippuryl-His-Leu hydrate

中文摘要：目的研究抗凝血肽的制备、分离方法及其体外抗凝血、溶栓效果。方法用木瓜蛋白酶、菠萝蛋白酶、中性蛋白酶和碱性蛋白酶对酪蛋白进行4酶混合水解制备抗凝血肽,以固定化凝血酶对抗凝血肽进行萃取,用新西兰兔进行体外抗凝血、溶栓实验。结果实验确定的抗凝血肽制备条件为酪蛋白质量浓度15%,水解每克酪蛋白的木瓜蛋白酶、菠萝蛋白酶、中性蛋白酶和碱性蛋白酶用量分别为1 500、2 400、1 000、1 250 U,水解温度50℃,pH 7.0,水解时间4 h。固定化凝血酶萃取抗凝血肽的初始浓度6 ATU/mL,萃取温度30℃,pH 5.0,萃取时间30 min。反萃取温度30℃,pH 7.6,反萃取时间40 min。抗凝血肽经高效体积排阻色谱分析,主要成分分子大小与马尿酰-组氨酰-亮氨酸相当。体外抗凝血时间超72 h,溶栓时间24 h。结论酪蛋白抗凝血肽主要成分为3肽,体外抗凝血及溶栓实验效果良好。

外文摘要：Objective To study the preparation and separation method for anticoagulant peptide and the effect of anticoagulation and thrombolysis in vitro.Methods The casein was hydrolyzed to prepare anticoagulant peptide using the mixed four enzymes such as papain,pineapple proteinase,neutral protease,and alkali protease.The anticoagulant peptide was extracted using immobilized thrombin.The effect of haemolysis and anticoagulation in vitro was investigated through the New Zealand rabbits experiments.Results The conditions of preparation anticoagulant peptide were as follows：quality of casein was 15%,papain proteinase,pineapple proteinase,neutral protease,and alcalase dosage were 1500,2400,1000,and 1250 U/（g casein）,respectively,temperature was 50 ℃,pH value was 7.0,and hydrolysis time was 4 h.The conditions for the extraction of anticoagulant peptide were as follows：the initial concentration was 6 ATU（Anti Thrombin Unit）/mL,temperature was 30 ℃,pH value was 5.0,and time was 30 min.Anti-extraction temperature was 30 ℃,pH value was 7.6,and time was 40 min.The purified anticoagulant peptide was analyzed via high performance size exclusion chromatography.The molecular weight of purified anticoagulant peptide was equal to N-Hippuryl-His-Leu hydrate and the main components were three peptides.The time of anticoagulation was more than 72 h and the time of hemolysis was 24 h in vitro.Conclusion The main components of anticoagulant peptides are three peptides.The effect of hemolysis and anticoagulation in vitro is good.