Van Staden Anton Du Preez, Faure Lindsay M, Vermeulen Ross R, Dicks Leon M T, Smith Carine
Department of Physiological Sciences , Stellenbosch University , Matieland 7602 , South Africa.
Department of Microbiology , Stellenbosch University , Matieland 7602 , South Africa.
ACS Synth Biol. 2019 Oct 18;8(10):2220-2227. doi: 10.1021/acssynbio.9b00167. Epub 2019 Sep 25.
Lanthipeptides are ribosomally synthesized and post-translationally modified peptides, with several having antimicrobial activity. The biosynthetic machinery responsible for modification of the class I lanthipeptide nisin provides a means for modification of a diverse range of lanthipeptides. However, literature regarding expression of class I lanthipeptides in a malleable Gram-negative host such as is limited. Here, we coexpressed precursor class I lanthipeptides fused to green fluorescent protein (GFP) along with the dehydratase and cyclase from the nisin operon. Fusion to GFP did not interfere with post-translational modifications as antimicrobially active nisin could be proteolytically liberated from the expressed GFP fusion. Additionally, we used this system to express two other class I lanthipeptides precursors fused to GFP (Pep5 and epilancin 15X), although only Pep5 exhibited consistent antimicrobial activity. This is the first report of a GFP-based fusion expression system for the expression of class I lanthipeptides in . The GFP-based fusion expression system is a robust system with the advantage of directly visualizing expression and purification through GFP fluorescence.
羊毛硫肽是核糖体合成并经翻译后修饰的肽，其中几种具有抗菌活性。负责修饰I类羊毛硫肽乳链菌肽的生物合成机制为多种羊毛硫肽的修饰提供了一种方法。然而，关于I类羊毛硫肽在可塑的革兰氏阴性宿主（如[此处原文缺失具体宿主名称]）中的表达的文献有限。在此，我们将与绿色荧光蛋白（GFP）融合的I类羊毛硫肽前体与来自乳链菌肽操纵子的脱水酶和环化酶共表达。与GFP融合并不干扰翻译后修饰，因为具有抗菌活性的乳链菌肽可以从表达的GFP融合物中通过蛋白水解作用释放出来。此外，我们使用该系统表达了另外两种与GFP融合的I类羊毛硫肽前体（Pep5和表皮兰辛15X），尽管只有Pep5表现出一致的抗菌活性。这是关于在[此处原文缺失具体宿主名称]中表达I类羊毛硫肽的基于GFP的融合表达系统的首次报道。基于GFP的融合表达系统是一个强大的系统，具有通过GFP荧光直接可视化表达和纯化的优点。
