Scripps Institution of Oceanography, University of California at San Diego, La Jolla, CA, USA.
Department of Pediatrics, University of California at San Diego, La Jolla, CA, USA.
Cell Chem Biol. 2019 May 16;26(5):724-736.e7. doi: 10.1016/j.chembiol.2019.02.004. Epub 2019 Mar 7.
The disconnect between the genomic prediction of secondary metabolite biosynthetic potential and the observed laboratory production profile of microorganisms is well documented. While heterologous expression of biosynthetic gene clusters (BGCs) is often seen as a potential solution to bridge this gap, it is not immune to many challenges including impaired regulation, the inability to recruit essential building blocks, and transcriptional and/or translational silence of the biosynthetic genes. Here we report the discovery, cloning, refactoring, and heterologous expression of a cryptic hybrid phenazine-type BGC (spz) from the marine actinomycete Streptomyces sp. CNB-091. Overexpression of the engineered spz pathway resulted in increased production and chemical diversity of phenazine natural products belonging to the streptophenazine family, including bioactive members containing an unprecedented N-formylglycine attachment. An atypical discrete adenylation enzyme in the spz cluster is required to introduce the formylglycine moiety and represents a phylogenetically distinct class of adenylation proteins.
次生代谢物生物合成潜能的基因组预测与微生物实验室观察到的生产情况之间存在脱节，这已得到充分证明。尽管异源表达生物合成基因簇（BGCs）通常被认为是弥合这一差距的一种潜在方法，但它并非没有许多挑战，包括调节受损、无法招募必要的构建块，以及生物合成基因的转录和/或翻译沉默。在这里，我们报告了从海洋放线菌链霉菌 sp. CNB-091 中发现、克隆、重构和异源表达一个隐藏的混合吩嗪型 BGC（spz）。工程化 spz 途径的过表达导致吩嗪天然产物（属于链霉菌吩嗪家族）的产量和化学多样性增加，包括含有前所未有的 N-甲酰甘氨酸附着的生物活性成员。spz 簇中一种非典型的离散氨酰化酶需要引入甲酰甘氨酸部分，代表了一个在系统发育上不同的氨酰化蛋白类群。
