New Cationic Antimicrobial Peptide Screened from Boiled-Dried Anchovies by Immobilized Bacterial Membrane Liposome Chromatography
New Cationic Antimicrobial Peptide Screened from Boiled-Dried Anchovies by Immobilized Bacterial Membrane Liposome Chromatography
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固定化细菌膜脂质体层析从水煮凤尾鱼中筛选新型阳离子抗菌肽
DOI：
10.1021/jf4052286
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发表时间：
2014-02-19
影响因子：
6.1
通讯作者：
Qian, Haifeng
中科院分区：
文献类型：
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作者：
Tang, Wenting;Zhang, Hui;Qian, Haifeng
采用固定化细菌膜脂质体色谱法筛选熟凤尾鱼干中潜在的抗菌肽。采用一步色谱法成功分离了一种新型阳离子抗菌肽Apep10。利用基质辅助激光解吸/电离四极杆飞行时间串联质谱（MALDI-Q-TOF MS）鉴定了Apep10的序列为GLARCLAGTL。抑菌活性评价表明，Apep10对对照菌（大肠杆菌、痢疾志贺菌、铜绿假单胞菌、鼠伤寒沙门菌、金黄色葡萄球菌、枯草芽孢杆菌和肺炎链球菌）的生长有抑制作用，最小抑菌浓度（MIC）为8 ~ 64 μ g/mL。浓度低于20 μ g/mL时，对小鼠红细胞几乎无细胞毒性。Apep10诱导的核苷酸渗漏表明该肽在细胞质膜上具有渗透活性。通过扫描电镜观察形态学变化。用碘化丙啶（PI）共聚焦激光扫描显微镜（CLSM）证实膜破坏。结果表明，固定化细菌膜脂质体层析法是一种从熟凤尾鱼中筛选具有细胞膜相互作用活性的未知抗菌肽的简单技术。
An efficient immobilized bacterial membrane liposome chromatography method was used to screen potential antimicrobial peptides from boiled-dried anchovies. A novel cationic antimicrobial peptide (Apep10) was successfully isolated by one-step chromatography. The sequence of Apep10 was identified as GLARCLAGTL by matrix-assisted laser desorption/ionization quadrupole time-of-flight tandem mass spectrometry (MALDI-Q-TOF MS). The antimicrobial activity assessment indicated that Apep10 inhibited the growth of the reference bacteria (Escherichia coli, Shigella dysenteriae, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Bacillus subtilis, and Streptococcus pneumoniae), with minimal inhibitory concentration (MIC) values ranging from 8 to 64 mu g/mL. Almost no cytotoxicity against mouse erythrocytes was observed at concentrations below 20 mu g/mL. Nucleotide leakage induced by Apep10 showed that the peptide exhibited permeable activity on the cytoplasmic membrane. Alterations in morphology were observed by scanning electronic microscopy (SEM). Membrane disruption was confirmed by confocal laser scanning microscopy (CLSM) with propidium iodide (PI). The results demonstrate that immobilized bacterial membrane liposome chromatography is a straightforward technique for screening unknown antimicrobial peptides with cell-membrane-interacting activities from boiled-dried anchovies.