Dr. Guangyu Yang
Project Description and Objectives
Directed evolution is a powerful tool for the engineering of enzymes. By mimicking natural evolution process in test tubes, random mutagenesis libraries of the target enzyme are created and then screened for desired properties. Directed evolution has been successful in improving enzyme activity, stability, and tolerance against organic solvent and substrate selectivity. However, since the libraries produced by random mutagenesis are extremely huge (e.g. by introducing 3 mutations in a protein of 200 amino acids, theoretically there would be 9 billion mutants), the success of enzyme direct evolution relies severely on the efficiency and accuracy of the screening method used. The traditional screening platforms, based on monitoring mutant activity on agar plates or in microtiter plates, are relatively low throughput (103~105/d) and laborious. Thus, screening method comprises the bottleneck of directed evolution and high throughput screening methods with high efficiency and high versatility are desired.
Integrate of multi-band fluorescence excitation
Detection optics in a microfluidic droplet screening chip
Establish a new droplet sorting mechanism that operates with high fidelity at a high droplet manipulation frequency
Validate the devices’ performance towards realizing the aimed on-chip MC-FADS for enzyme directed evolution.