五谷虫抑菌活性多肽RP-HPLC分离与纯化方法研究  

A Study on Separation and Purification of Antibacterial Peptides in Maggot with RP-HPLC

作　　者：王敏[1] 褚夫江[1] 巫春旭 刘文彬[1] 金小宝[1] 朱家勇[1] 

机构地区：[1]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,广东广州510006

出　　处：《江西农业大学学报》2017年第3期567-571,共5页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家自然科学基金项目(81274061)~~

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。This study aims at establishing a method for separation ol antibacterial peptides Irom Maggot and providing experimental evidence lor lurther study on its medicinal value. Peptides,with molecular weight below 30ku were obtained through the process of homogenization, filtrating, heating, centrifuging and ultrafiltra-tion. Subsequently, the method of SuperdexTM30 gel filtration chromatography was utilized to separate peptides and to identified the antibacterial activity peaks,which were lurther purilied with RP-HPLC.The antibacterial components ol S2 and S3 were screened out with gel liltration chromatography. Besides, seven peaks in S2 andlour peaks in S3 were identilied with RP-HPLC.The RSD ol retention time and peak area were all less than 5 %in stability,precision and repetition experiment, and the similarity ol 6 bathes ol samples was more than 0.99. The method ol RP-HPLC can be used to quickly separate antibacterial polypeptides lrom Maggot, and it is sta- ble,reliable and reproducible.

关 键 词：五谷虫 抑菌肽 RP-HPLC 分离 纯化 

分 类 号：R384.2[医药卫生—医学寄生虫学] Q966[医药卫生—基础医学]