Exploring sequence space in search of functional enzymes using microfluidic droplets

This is the author accepted manuscript. The final version is available from Elsevier via the DOI in this record.Screening of enzyme mutants in monodisperse picoliter compartments, generated at kilohertz speed in microfluidic devices, is coming of age. After a decade of proof-of-principle experiments, workflows have emerged that combine existing microfluidic modules to assay reaction progress quantitatively and yield improved enzymes. Recent examples of the screening of libraries of randomised proteins and from metagenomic sources suggest that this approach is not only faster and cheaper, but solves problems beyond the feasibility scope of current methodologies. The establishment of new assays in this format - so far covering hydrolases, aldolases, polymerases and dehydrogenases - will enable the exploration of sequence space for new catalysts of natural and non-natural chemical transformations.This work was funded by the Engineering and Physical Sciences Research Council (EPSRC; studentship in the Centre for Doctoral Training “Sensor Technologies and Applications” to P.M., EP/L015889/1) and the Biotechnology and Biological Research Council (BBSRC; BB/K013629/1)