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
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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-HPKLC)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～225nm短波紫外区，AMP-1有紫外吸收。
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关键词:
- 小皱蝽,抗菌肽,纯化,HPLC
Abstract: 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 C1s 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 inhinhibition assay on S. aureus and E. coli, AMP- 1 is the antimicrobial polypeptide. -
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