期刊
CELL REPORTS
卷 20, 期 5, 页码 1029-1037出版社
CELL PRESS
DOI: 10.1016/j.celrep.2017.07.015
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资金
- European Union Seventh Framework Programme [FP7-KBBE-2013-7-single-stage, 613745]
- Novo Nordisk Foundation (NNF) [11355-444]
- NNF [NNF16CC0020908]
- NNF
- Lundbeck Foundation [R140-2013-13496, R77-2010-6772] Funding Source: researchfish
- NNF Center for Biosustainability [Bacterial Synthetic Biology] Funding Source: researchfish
- Novo Nordisk Fonden [NNF10CC1016517, NNF13SA0009311, NNF13SA0006019, NNF14OC0011335] Funding Source: researchfish
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.
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