A microfluidic platform is utilized for rapid enzyme engineering relevant to identify improved and/or missing catalysts in production pathways. This approach is compatible with aerobic/anaerobic enzyme screening (in vivo or cell-free). The system is capable of utilizing TF-sensors or FRET and enzyme-linked sensors. Once candidate genes have been identified, variants are synthesized using rational design/directed evolution approaches. Appropriate sensors are linked to monitor reaction improvements, and in-depth characterization of candidate enzymes is carried out in vitro and in target host systems. The pairing of microfluidics and biosensor design/engineering for molecular adaptation is only possible in a few unique research environments. It can be used to eliminate bottlenecks in metabolic engineering. Improved biocatalysts can be incorporated into production strains to improve titers, rates, and yields of bioproducts. This approach is aided by the availability of biosensors and is most successful with target proteins where (i) structures can be inferred from database resources or (ii) functional correlations between active and inactive enzymes can result from the study of evolutionary relationships in sequences of large numbers of homologs. Ready to use through collaboration. The strategy works best if able to be approached with a structural perspective or enzymes from large structural or functional classes.
Argonne National Laboratory
Los Alamos National Laboratory
National Renewable Energy Laboratory