Synthetic DNA Libraries for Protein Engineering Toward Process Improvement in Drug Synthesis

Speeding-up enzyme engineering by directed evolution is a primary target to be achieved for a wider uptake of biocatalysis in pharmaceutical process development. The capability to rapidly generate the designed sequence diversity has profound implications in the overall optimization of protein function. Drawbacks associated with traditional PCR methods for sequence diversification interfere with the generation of all the variants that have been designed. On the contrary, the enhanced quality of synthetic DNA libraries makes the exploration of sequence space more efficient. Here, methods for the effective utilization of synthetic DNA libraries are described. The overall procedure allows the generation of ready-to-screen libraries within two weeks from synthetic DNA acquisition.