The goal of this project is to develop a new approach to facilitate the development of faster, greener, and cheaper ways of synthesizing drugs. The challenge in using enzymes is that natural occurring enzymes do not often catalyze the exact reaction that is needed for a pharmaceutical development project. The goal in this project is to develop a high-throughput, highly miniaturized method of engineering enzymes to perform desired reactions. It is anticipated that this technology can be used to develop such biocatalysts in a few days for any desired chemical transformation. This capability could greatly facilitate drug development projects by making available tailor-made catalysts for any reaction in a time and cost-effective manner.
Enzyme engineering for pharmaceutical process chemistry and manufacturing has become a revolutionary technology as enzymatic transformations potentially offer high yield, selectivity and enantiomeric purity with low cost and green chemistry. The team proposes development of a novel screening system to reduce the material use, eliminate special reagents and improve the overall throughput in enzyme engineering and screening. The three key modules include (i) cell-free protein synthesis (CFPS), (ii) droplet microfluidics and (iii) mass spectrometry (MS). CFPS offers the potential speed advantage as it bypasses the cell growth and protein harvest steps. It can be completed with shorter cycle time, produces purer proteins, and at the same time allows active monitoring, direct manipulation, and rapid sampling. Droplet microfluidics offers a platform for performing reactions and assays in nanoliter or less volume which is also adaptive to be automated. MS allows the possibility for fast development of label-free assays for a wide variety of potential substrates with minimal delay. The proposed approach is transformative because it will allow virtually any enzymatic transformation to be engineered in 10-fold less time by eliminating the need to develop labels, coupled reactions, or using cells in the process. The proposed technology will be useful also for other applications in chemical process development. This award by the Biotechnology and Biochemical Engineering Program of the CBET Division is co-funded by the GOALI Program of the Division of Industrial Innovation and Partnerships.
