Abstract
An extension of directed evolution strategies to genome-wide variations increases the chance of obtaining metabolite-overproducing microbes. However, a general high-throughput screening platform for selecting improved strains remains out of reach. Here, to expedite the evolution of metabolite-producing microbes, we utilize synthetic RNA devices comprising a riboswitch and a selection module that specifically sense inconspicuous metabolites. Using L-lysine-producing Escherichia coli as a model system, we demonstrated that this RNA device could enrich pathway-optimized strains to up to 75% of the total population after four rounds of enrichment cycles. Furthermore, the potential applicability of this device was examined by successfully extending its application to the case of L-tryptophan. When used in conjunction with combinatorial mutagenesis for metabolite overproduction, our synthetic RNA device should facilitate strain improvement.
| Original language | English |
|---|---|
| Article number | 1413 |
| Journal | Nature Communications |
| Volume | 4 |
| DOIs | |
| State | Published - 2013 |
Bibliographical note
Funding Information:This work was supported by grants from the Advanced Biomass R&D Center (ABC) (ABC-2010-0029800) and World Class University (WCU) program (R31-10105) of the National Research Foundation of Korea funded by the Ministry of Education, Science and Technology and by the Marine Biotechnology Program and the Manpower Development Program funded by Ministry of Land, Transport and Maritime Affairs of Korean Government. We thank Dr J.B. Seo of KSBI for the analysis of intracellular lysine concentration.