Qi Yunci, Ding Edward, Blodgett Joshua A V
Department of Biology, Washington University in St Louis , St Louis, Missouri 63130, United States.
ACS Synth Biol. 2018 Feb 16;7(2):357-362. doi: 10.1021/acssynbio.7b00349. Epub 2017 Dec 20.
Polycyclic tetramate macrolactam (PTM) natural products are produced by actinomycetes and other bacteria. PTMs are often bioactive, and the simplicity of their biosynthetic clusters make them attractive for bioengineering. Clifednamide-type PTMs from Streptomyces sp. strain JV178 contain a distinctive ketone group, suggesting the existence of a novel PTM oxidizing enzyme. Here, we report the new cytochrome P450 enzyme (CftA) is required for clifednamide production. Genome mining was used to identify several new clifednamide producers, some having improved clifednamide yields. Using a parallel synthetic biology approach, CftA isozymes were used to engineer the ikarugamycin pathway of Streptomyces sp. strain NRRL F-2890 to yield clifednamides. Further, we observed that strong CftA expression leads to the production of a new PTM, clifednamide C. We demonstrate the utility of both genome mining and synthetic biology to rapidly increase clifednamide production.
多环四酰胺大环内酯（PTM）天然产物由放线菌和其他细菌产生。PTM通常具有生物活性，其生物合成簇的简单性使其在生物工程方面具有吸引力。来自链霉菌属菌株JV178的克利夫地酰胺型PTM含有一个独特的酮基，这表明存在一种新型的PTM氧化酶。在此，我们报道克利夫地酰胺的产生需要新的细胞色素P450酶（CftA）。通过基因组挖掘来鉴定几个新的克利夫地酰胺产生菌，其中一些具有提高的克利夫地酰胺产量。使用平行合成生物学方法，利用CftA同工酶对链霉菌属菌株NRRL F-2890的伊卡鲁霉素途径进行工程改造以产生克利夫地酰胺。此外，我们观察到CftA的强表达会导致产生一种新的PTM，即克利夫地酰胺C。我们展示了基因组挖掘和合成生物学在快速提高克利夫地酰胺产量方面的效用。
