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

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.