Genetic Selection for Small Molecule Production in Competitive Microfluidic Droplets.
ACS Synth Biol
; 8(8): 1737-1743, 2019 08 16.
Article
in En
| MEDLINE
| ID: mdl-31356044
ABSTRACT
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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Biosensing Techniques
/
Coculture Techniques
/
Microfluidics
Language:
En
Journal:
ACS Synth Biol
Year:
2019
Document type:
Article
Affiliation country:
United States