Microfluidic droplets as tools for high-throughput biology: Enzyme evolution, recruitment and discovery based on catalytic promiscuity

Conference Dates

September 24-28, 2017


‘Promiscuous’ enzymes possess additional activities in addition to their native ones, challenging the textbook adage “one enzyme – one activity”. The observation of strong promiscuous activities in the alkaline phosphatase (AP) superfamily - where one active site can catalyse up to six chemically distinct hydrolytic reactions with promiscuous second order rate accelerations between 10e9 and 10e17 - suggests that even broadly promiscuous catalysis can be rather efficient. To efficiently explore the interconversion of promiscuous enzyme, we use picoliter water-in–oil emulsion droplets produced in microfluidic devices as high-throughput screening reactors. We present new workflows that allow screening of >10e6 clones and permit successful selections from single protein and metagenomic libraries, where lower throughput approaches have failed.

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