Synthetic RNA devices to expedite the evolution of metabolite-producing microbes.
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.
Department of Chemical Engineering, Pohang University of Science and Technology, San 31, Hyoja-dong, Nam-gu, Pohang 790-784, Korea.
Jina
J
Yang
Sang Woo
SW
Seo
Sungho
S
Jang
So-I
SI
Shin
Chae Hyun
CH
Lim
Tae-Young
TY
Roh
Gyoo Yeol
GY
Jung
D001483:Base Sequence; D019020:Directed Molecular Evolution / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism*; D008239:Lysine / Q000096:biosynthesis; D053858:Metabolic Networks and Pathways; D055442:Metabolome; D008954:Models, Biological; D008969:Molecular Sequence Data; D012313:RNA / Q000378:metabolism*; D014364:Tryptophan / Q000378:metabolism
D012313:RNA; D014364:Tryptophan; D008239:Lysine