Automated high throughput workflow for rapid implementation of immobilized enzymes in chemical process development

Abstract

Despite known advantages of immobilized enzymes, their broad application in pharmaceutical drug manufacturing has not yet caught up. Limited access to commercially available immobilized enzymes on one side and time – and cost – consuming development of new processes on the other side have hampered their broader implementation so far. Herein, we present a walk-away high throughput workflow with state of the art robotic equipment that enables the rapid optimization and development of immobilized enzymes – ready to be scaled - and thus matching the need for speed in modern drug development. Following this workflow, we were able to identify an engineered ketoreductase which was >2-fold more active than the wildtype from L. kefir. Further optimization of the reaction and immobilization conditions lead to an additional 29-fold productivity increase from 0.36 to 10.6 g_product g_enzyme⁻¹ for the selected enzyme variant.