Recombinant expression and purification of the antimicrobial peptide magainin‐2
Recombinant expression and purification of the antimicrobial peptide magainin‐2
复制标题
DOI：
10.1002/btpr.1650
复制
发表时间：
2013
影响因子：
2.9
通讯作者：
R. Ramos;S. Moreira;A. Rodrigues;M. Gama;L. Domingues
中科院分区：
文献类型：
--
作者：
R. Ramos;S. Moreira;A. Rodrigues;M. Gama;L. Domingues
Magainin-2（MAG 2）是一种从非洲爪蟾皮肤中分离的多聚阳离子抗菌肽。它对革兰氏阳性和革兰氏阴性细菌、真菌具有广谱抗菌活性，并诱导原生动物的渗透溶解。MAG 2还具有抗病毒和抗肿瘤特性。这些活性使该肽成为治疗应用的有希望的候选物。重组表达系统对于负担得起的大量生物活性肽的生产是必需的。在这项工作中，MAG 2已被克隆到与来自热纤梭菌的接头序列（LK-CBM 3）融合的III家族碳水化合物结合模块的N末端;在两个模块之间引入了一个甲酸识别位点，用于化学切割肽。在大肠杆菌BL 21（DE 3）中表达了重组蛋白MAG 2-LK-CBM 3，并成功地从融合伴侣LK-CBM 3中切割并纯化了MAG 2。通过测试其对革兰氏阴性菌的活性来证实其功能性。© 2012美国化学工程师学会生物技术。程序：2013
Magainin‐2 (MAG2) is a polycationic antimicrobial peptide isolated from the skin of the African clawed frog Xenopus laevis. It has a wide spectrum of antimicrobial activities against gram‐positive and gram‐negative bacteria, fungi, and induces osmotic lysis of protozoa. MAG2 also possesses antiviral and antitumoral properties. These activities make this peptide a promising candidate for therapeutic applications. Recombinant expression systems are necessary for the affordable production of large amounts of the biologically active peptide. In this work, MAG2 has been cloned to the N‐terminal of a family III carbohydrate‐binding module fused to the linker sequence (LK‐CBM3) from Clostridium thermocellum; a formic acid recognition site was introduced between the two modules for chemical cleavage of the peptide. The recombinant protein MAG2‐LK‐CBM3 was expressed in Escherichia coli BL21 (DE3) and MAG2 was successfully cleaved and purified from the fusion partner LK‐CBM3. Its functionality was confirmed by testing its activity against gram‐negative bacteria. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2013