作 者：
王敏;褚夫江;巫春旭;刘文彬;金小宝;朱家勇
单 位：
广东药科大学基础学院药用生物活性物质研究所/广东省生物活性药物研究重点实验室
关键词：
五谷虫;抑菌肽;RP-HPLC;分离;纯化
摘 要：
旨在建立一种五谷虫抑菌活性多肽分离纯化的方法,为研究其药用价值提供实验依据。通过匀浆、过滤、加热、离心后利用超滤膜超滤获得分子量在30 ku以下的蛋白多肽,采用凝胶过滤层析系统初步分离并筛选出抑菌活性峰,以RP-HPLC建立该活性峰的分离纯化方法。结果显示,凝胶过滤层析法分离并且筛选出两个具有抑菌活性组分S2和S3;RP-HPLC色谱法确定了S2具有7个峰和S3具有4个峰;该方法稳定性、精密度、重复性实验的保留时间与峰面积的RSD值均小于5%;6批样品RP-HPLC图谱相似度值大于0.99;采用RPHPLC能快速分离五谷虫抑菌活性多肽,且该方法稳定、可靠、重复性好。
译 名：
A Study on Separation and Purification of Antibacterial Peptides in Maggot with RP-HPLC
作 者：
WANG Min;CHU Fu-jiang;WU Chun-xu;LIU Wen-bin;JIN Xiao-bao;ZHU Jia-yong;Institution of Pharmaceutical Bioactive Substances,School of Basic Courses,Guangdong Pharmaceutical University/Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances;
关键词：
Maggot;;antibacterial peptides;;RP-HPLC;;separation
摘 要：
This study aims at establishing a method for separation of antibacterial peptides from Maggot and providing experimental evidence for further study on its medicinal value. Peptides,with molecular weight below 30 ku were obtained through the process of homogenization,filtrating,heating,centrifuging and ultrafiltration.Subsequently,the method of SuperdexTM30 gel filtration chromatography was utilized to separate peptides and to identified the antibacterial activity peaks,which were further purified with RP-HPLC. The antibacterial components of S2 and S3 were screened out with gel filtration chromatography.Besides,seven peaks in S2 and four peaks in S3 were identified with RP-HPLC.The RSD of retention time and peak area were all less than 5%in stability,precision and repetition experiment,and the similarity of 6 bathes of samples was more than 0.99.The method of RP-HPLC can be used to quickly separate antibacterial polypeptides from Maggot,and it is stable,reliable and reproducible.
