期刊
ACS SYNTHETIC BIOLOGY
卷 8, 期 8, 页码 1737-1743出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.9b00226
关键词
microfluidics; biosensors; coculture; metabolic engineering; muconate; lignin
资金
- Laboratory Directed Research and Development program at the Oak Ridge National Laboratory
- DOE [DE-AC05-000R22725]
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.
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