High-throughput Discovery Lab

The application of enzymes to catalyze industrial reactions, known as “biocatalysis”, has the potential to make indsutrial chemical processes less contaminant and more sustainable, following the principles of Green Chemistry. The natural diversity represents a wealthy source of microbial biocatalysts, which can be mined using functional metagenomics and tailored using directed evolution.  In either case, finding a suitable enzyme for the desired reaction requires exploring vast genetic diversity in timeframes compatible with industrial process development and the screening of metagenomic or directed evolution libraries has been particularly hindered by the lack of ultrahigh-throughput screening methods that are independent of complex and expensive equipment. Using “droplet” microfluidics, enzyme assays are miniaturized to the size of a picoliter droplet with an increase in throughput as well as a reduction of assay costs, making droplet-based screening the best option to screen large libraries of enzymes or enzyme variants. 

This TFM project deals with development of ultrahigh-throughput screening assays using in vitro (using cell-free protein expression) or in vivo (using microbial hosts for recombinant expression) for future implementation in microfluidic droplets to carry out directed evolution or functional metagenomics campaigns.  Using a model enzyme, conditions for protein expression coupled with the enzymatic assay will be optimized in solution.  Subsequently, depending on progress in the previous stage, these assays may be adapted to function within water-in-oil microdroplets and optimized.