Investigation of the Biosynthesis of the Lasso Peptide Chaxapeptin Using an E. coli-Based Production System.
Investigation of the Biosynthesis of the Lasso Peptide Chaxapeptin Using an E. coli-Based Production System.
复制标题
DOI：
10.1021/acs.jnatprod.8b00392
复制
发表时间：
2018-09
影响因子：
5.1
通讯作者：
Helena Martin-Gómez;U. Linne;F. Albericio;Judit Tulla-Puche;J. Hegemann
中科院分区：
文献类型：
--
作者：
Helena Martin-Gómez;U. Linne;F. Albericio;Judit Tulla-Puche;J. Hegemann
Lasso肽是属于核糖体合成和后修饰肽（RIPPs）家族的天然产物，并且由其独特的拓扑结构定义。尽管在许多不同种类的细菌中发现了套索肽生物合成基因簇，但迄今为止研究的大多数套索肽是蛋白细菌或放线菌来源的。尽管如此，没有E。已经报道了基于大肠杆菌的生产系统用于放线菌套索肽，而对于变形菌套索肽有许多这样的例子。在此，在E.杆菌Chaxapeptin最初分离自Streptomyces leeuwenhoekii菌株C58，与套索肽sungsanpin（由海洋链霉菌产生）密切相关。并具有抑制人肺癌细胞系A549细胞侵袭的活性。我们的生产系统不仅允许在天然生产者之外以0.1mg/L的产率分离成熟套索肽（与来自S. Leeuwenhoekii），但也用于突变研究以鉴定前体肽中对生物合成重要的残基。除了这些实验之外，还评估了chaxapeptin对热变性和蛋白酶的稳定性。
Lasso peptides are natural products belonging to the family of ribosomally synthesized and posttranslationally modified peptides (RiPPs) and are defined by their unique topology. Even though lasso peptide biosynthetic gene clusters are found in many different kinds of bacteria, most of the hitherto studied lasso peptides were of proteobacterial or actinobacterial origin. Despite this, no E. coli-based production system has been reported for actinobacterial lasso peptides, while there are numerous examples of this for proteobacterial lasso peptides. Here, a heterologous production system of the lasso peptide chaxapeptin was established in E. coli. Chaxapeptin, originally isolated from Streptomyces leeuwenhoekii strain C58, is closely related to the lasso peptide sungsanpin (produced by a marine Streptomyces sp.) and shares its inhibitory activity against cell invasion by the human lung cancer cell line A549. Our production system not only allowed isolation of the mature lasso peptide outside of the native producer with a yield of 0.1 mg/L (compared to 0.7 mg/L from S. leeuwenhoekii) but also was used for a mutational study to identify residues in the precursor peptide that are important for biosynthesis. In addition to these experiments, the stability of chaxapeptin against thermal denaturation and proteases was assessed.