高效液相色谱法分离纯化小皱蝽抗菌肽AMP-1  

PURIFICATION OF THE ANTIMICROBIAL PEPTIDE AMP-1 OF CYCLOPELTA PARVA DISTANT BY HPIC

作　　者：杨泽宏[1] 杨星勇[1] 袁小红[1] 罗克明[1] 罗小英[1] 张永军[1] 郭余龙[1] 裴炎[1] 

机构地区：[1]西南农业大学生物技术中心,重庆400716

出　　处：《西南农业大学学报（自然科学版）》2001年第2期104-107,共4页Journal of Southwest Agricultural University

基　　金：重庆市科委攻关项目;西南农业大学专项基金项目

摘　　要：用昆虫生理盐水浸提小皱蝽成虫 ,浸提液冻干后 ,酸提取法制备粗提液 ,固相萃取 (SPE)初分级分离 ,活性洗脱组分反相高效液相色谱 (RP -HPLC)纯化 ,进一步凝胶渗透色谱 (GPC)及反相高效液相色谱 (RP -HPLC)纯化 ,分离得到一抗菌肽 ,茚三酮反应黄色 ,命名为AMP - 1。在 190nm～ 2 2 5nm短波紫外区 ,AMP - 1有紫外吸收。An antimicrobial peptide,AMP-1,was isolated from the bug Cyclopelta parva Distant,an insect of Pentatomidae,Hemiptera,by the extraction and preparation of the crude extract with the acid extraction methods and fractionation of Sep-Pak C 18 Cartridge.Subsequently,40% Sep-Pak fraction was purified by the initial reverse phase HPLC.The active peak was further fractionated by gel-permeation chromatography HPLC and the final reverse phase HPLC.All the HPLC purifications utilized the Acetonitrile-Water-Trifluoroacetic acetic elution system as mobile phase,with detection at 225 nm.The pure substances were recovered and the anti-bacterial activity of each aliquot was routinely monitored by the plate growth inhibition assay on S.aureus and E.coli, AMP-1 is the antimicrobial polypeptide.

关 键 词：小皱蝽 抗菌肽 纯化 HPLC 

分 类 号：Q966[生物学—昆虫学]