Native and Engineered Clifednamide Biosynthesis in Multiple Streptomyces spp.
Native and Engineered Clifednamide Biosynthesis in Multiple Streptomyces spp.
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DOI：
10.1021/acssynbio.7b00349
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发表时间：
2018-02-01
影响因子：
4.7
通讯作者：
Blodgett, Joshua A. V.
中科院分区：
文献类型：
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作者：
Qi, Yunci;Ding, Edward;Blodgett, Joshua A. V.
多环四酸大内酰胺(PTM)天然产物是由放线菌等细菌产生的。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中具有吸引力。链霉菌Clive dnamide-type PTMS的研究菌株JV178含有一个独特的酮基，提示存在一种新的PTM氧化酶。在这里，我们报告了新的细胞色素P450酶(CFTA)是生产克利福那胺所必需的。基因组挖掘被用来确定几个新的克立非那胺生产商，其中一些已经提高了克立非那胺的产量。采用平行合成生物学方法，利用CFTA同工酶对链霉菌的伊卡鲁霉素途径进行了基因工程研究。菌株NRRL F-2890产生克立苯那胺。此外，我们观察到CFTA的强表达导致了一种新的PTM的产生，clfednamide C。我们证明了基因组挖掘和合成生物学的效用，以快速增加clfednamide的产量。
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.