Genetic-Metabolic Coupling for Targeted Metabolic Engineering
Genetic-Metabolic Coupling for Targeted Metabolic Engineering
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DOI：
10.1016/j.celrep.2017.07.015
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发表时间：
2017-08-01
期刊：
影响因子：
8.8
通讯作者：
Sommer, Morten Otto Alexander
中科院分区：
文献类型：
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作者：
Cardinale, Stefano;Tueros, Felipe Gonzalo;Sommer, Morten Otto Alexander
微生物中化学物质的生产通常使用强大的生物合成酶，这些酶可以与微生物的天然代谢相互作用，从而影响细胞的适应性和产品的产量。然而，生产优化在很大程度上依赖于在没有代谢紊乱的情况下从野生型菌株收集的数据，从而限制了它们与特定条件的相关性。在这里，我们通过使用合成的RNA生物传感器将细胞适应性与大肠杆菌中硫胺素二磷酸的生产偶联来解决这个问题。我们使用这种策略来询问一个转座子突变体库，并阐明影响细胞适应度和硫胺素产生的天然基因网络。最后，我们确定了OxyR-Fur胁迫反应的效应物，这些效应物通过铁储存和Fe-S簇在酶中的替代调节来限制硫胺素的生物合成。这项研究提出了一种可靠的高通量鉴定已知和未知功能的遗传靶点的新方法，这些靶点与特定的生物合成过程直接相关。
Production of chemicals in microbes often employs potent biosynthetic enzymes, which can interact with the microbial native metabolism to affect cell fitness and product yield. However, production optimization largely relies on data collected from wild-type strains in the absence of metabolic pertur-bations, thus limiting their relevance to specific conditions. Here, we address this issue by coupling cell fitness to the production of thiamine diphosphate in Escherichia coli using a synthetic RNA biosensor. We use this strategy to interrogate a library of transposon mutants and elucidate the native gene network influencing both cell fitness and thiamine production. Ultimately, we identify effectors of the OxyR-Fur stress response that limit thiamine biosynthesis via alternative regulation of iron storage and Fe-S cluster inclusion in enzymes. This study presents a new approach for the reliable high-throughput identification of genetic targets of both known and unknown function that are directly relevant to a specific biosynthetic process.