Journal article
Genetic-Metabolic Coupling for Targeted Metabolic Engineering
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
Bacterial Synthetic Biology, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
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.
Language: | English |
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Publisher: | Elsevier |
Year: | 2017 |
Pages: | 1029-1037 |
ISSN: | 22111247 |
Types: | Journal article |
DOI: | 10.1016/j.celrep.2017.07.015 |
ORCIDs: | Cardinale, Stefano , Tueros Farfan, Felipe Gonzalo and Sommer, Morten Otto Alexander |
Bacterial Proteins Biology (General) Escherichia coli Escherichia coli Proteins Fe-S clusters Metabolic Engineering QH301-705.5 RNA biosensors Repressor Proteins Thiamine Pyrophosphate ferric uptake regulating proteins, bacterial oxidative stress response oxyR protein, E coli thiamine biosynthesis transposon mutagenesis