Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210046, China.
Microbiol Res. 2010 Oct 20;165(8):713-8. doi: 10.1016/j.micres.2010.01.002. Epub 2010 Jan 20.
Different strategies have been developed to produce small antimicrobial peptides using recombinant techniques. Here we report a new technology of biosynthesis of moricin CM4 and human β-defensins 4 (HβD4) in the Escherichia coli. The CM4 and HβD4 gene were cloned into a vector containing the tags elastin-like peptide (ELP) and intein to construct the expression vector pET-EI-CM4 and pET-EI-HβD4. All the peptides, expressed as soluble fusions, were isolated from the protein debris by the method called inverse transition cycling (ITC) rather than traditional immobilized metal affinity chromatography (IMAC) and separated from the fusion leader by self-cleavage. Fully reduced peptides that were purified exhibited expected antimicrobial activity. The approach described here is a low-cost, convenient and potential way for generating small antimicrobial peptide.
已经开发了不同的策略来使用重组技术生产小的抗菌肽。在这里，我们报告了一种在大肠杆菌中生物合成莫瑞西菌素 CM4 和人 β-防御素 4 (HβD4) 的新技术。CM4 和 HβD4 基因被克隆到一个含有弹性蛋白样肽 (ELP) 和内含子标签的载体中，构建了表达载体 pET-EI-CM4 和 pET-EI-HβD4。所有的肽，作为可溶融合蛋白表达，通过称为逆相变循环（ITC）的方法而不是传统的固定化金属亲和层析（IMAC）从蛋白碎片中分离出来，并通过自切割从融合前导肽中分离出来。完全还原的肽经纯化后表现出预期的抗菌活性。这里描述的方法是一种低成本、方便且具有潜力的生成小抗菌肽的方法。
