Genetic Selection for Small Molecule Production in Competitive Microfluidic Droplets

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 physica...

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Bibliographic Details
Published in:ACS synthetic biology 2019-08, Vol.8 (8), p.1737-1743
Main Authors: Millet, Larry J, Vélez, Jessica M, Michener, Joshua K
Format: Article
Language:English
Subjects:
BASIC BIOLOGICAL SCIENCES
biosensors
coculture
lignin
metabolic engineering
microfluidics
muconate
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Summary: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. 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. Facile selections for small molecule production will increase testing throughput for engineered microbes and allow for the rapid optimization of novel metabolic pathways.
ISSN:2161-5063
2161-5063
DOI:10.1021/acssynbio.9b00226