Journal
CHEMISTRY & BIOLOGY
Volume 19, Issue 8, Pages 1001-1009Publisher
CELL PRESS
DOI: 10.1016/j.chembiol.2012.06.009
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Funding
- Research Councils UK Basic Technology Grant
- European Union (EU)
- Biotechnology and Biological Sciences Research Council
- EU Marie-Curie Programme
- Natural Sciences and Engineering Research Council of Canada
- Engineering and Physical Sciences Research Council [EP/H046593/1, EP/D048664/1] Funding Source: researchfish
- EPSRC [EP/D048664/1, EP/H046593/1] Funding Source: UKRI
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We demonstrate the utility of a microfluidic platform in which water-in-oil droplet compartments serve to miniaturize cell lysate assays by a million-fold for directed enzyme evolution. Screening hydrolytic activities of a promiscuous sulfatase demonstrates that this extreme miniaturization to the single-cell level does not come at a high price in signal quality. Moreover, the quantitative readout delivers a level of precision previously limited to screening methodologies with restricted throughput. The sorting of 3 x 10(7) monodisperse droplets per round of evolution leads to the enrichment of clones with improvements in activity (6-fold) and expression (6-fold). The detection of subtle differences in a larger number of screened clones provides the combination of high sensitivity and high-throughput needed to rescue a stalled directed evolution experiment and make it viable.
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