Genetic Selection for Small Molecule Production in Competitive Microfluidic Droplets

Millet, Larry J.; Velez, Jessica M.; Michener, Joshua K.

doi:10.1021/acssynbio.9b00226

Title: Genetic Selection for Small Molecule Production in Competitive Microfluidic Droplets

Journal Article · 29 July 2019 · ACS Synthetic Biology

DOI: https://doi.org/10.1021/acssynbio.9b00226 · OSTI ID:1550760

^[1]; Velez, Jessica M. ^[1];

^[2]

Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Tennessee, Knoxville, TN (United States)
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

Biosensors can be used to screen or select for small molecule production in engineered microbes. However, mutations to the biosensor that interfere with accurate signal transduction are common, producing an excess of false positives. Strategies have been developed to avoid this limitation by physically separating the production pathway and biosensor, but these approaches have only been applied to screens, not selections. In this work, we have developed a novel biosensor-mediated selection strategy using competition between cocultured bacteria. When applied to the biosynthesis of cis,cis-muconate, we show that this strategy yields a selective advantage to producer strains that outweighs the costs of production. By encapsulating the competitive cocultures into microfluidic droplets, we successfully enriched the muconate-producing strains from a large population of control nonproducers. Finally, facile selections for small molecule production will increase testing throughput for engineered microbes and allow for the rapid optimization of novel metabolic pathways.

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Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: USDOE

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 1550760

Journal Information:: ACS Synthetic Biology, Vol. 8, Issue 8; ISSN 2161-5063

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 5 works

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