| Autor: |
Millet LJ; Biosciences Division , Oak Ridge National Laboratory , Oak Ridge , Tennessee 37830 , United States.; The Joint Research Activity Unit of The Bredesen Center for Interdisciplinary Research and Graduate Education , University of Tennessee , Knoxville , Tennessee 37996 , United States., Vélez JM; Biosciences Division , Oak Ridge National Laboratory , Oak Ridge , Tennessee 37830 , United States.; The Bredesen Center for Interdisciplinary Research and Graduate Education , University of Tennessee , Knoxville , Tennessee 37996-3394 , United States., Michener JK; Biosciences Division , Oak Ridge National Laboratory , Oak Ridge , Tennessee 37830 , United States. |
| Abstrakt: |
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. |
