title: Exploring sequence space in search of functional enzymes using microfluidic droplets.
abstract: 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.
authors.affiliation: Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge CB2 1GA, UK.
authors.forename: Philip
authors.initials: P
authors.lastname: Mair
authors.affiliation: Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge CB2 1GA, UK; Living Systems Institute, University of Exeter, Stocker Road, Exeter EX4 4QD, UK.
authors.forename: Fabrice
authors.initials: F
authors.lastname: Gielen
authors.affiliation: Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge CB2 1GA, UK. Electronic address: fh111@cam.ac.uk.
authors.forename: Florian
authors.initials: F
authors.lastname: Hollfelder
mesh_terms: D057075:Enzyme Assays / Q000379:methods*; D004798:Enzymes / Q000235:genetics / Q000378:metabolism*; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D009154:Mutation
chemical_list: D004798:Enzymes