PT  - JOURNAL ARTICLE
AU  - Stefano Cardinale
AU  - Felipe Gonzalo Tueros
AU  - Morten Otto Alexander Sommer
TI  - Genetic-metabolic coupling for targeted metabolic engineering
AID  - 10.1101/156927
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 156927
4099  - http://biorxiv.org/content/early/2017/07/02/156927.short
4100  - http://biorxiv.org/content/early/2017/07/02/156927.full
AB  - To produce chemicals, microbes typically employ potent biosynthetic enzymes that interact with native metabolism to affect cell fitness as well as chemical production. However, production optimization largely relies on data collected from wild type strains in the absence of metabolic perturbations, thus limiting their relevance to specific process scenarios. Here, we address this issue by coupling cell fitness to the production of thiamine diphosphate in Escherichia coli using a synthetic RNA biosensor.We apply this system to interrogate a library of transposon mutants to elucidate the native gene network influencing both cell fitness and thiamine production. Specifically, we identify uncharacterized effectors of the OxyR-SoxR stress response that limit thiamine biosynthesis via alternative regulation of iron storage and Fe-S-cluster inclusion in enzymes.Our study represents a new generalizable 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.