Directed Evolution in Femtolitre Microreactors Exploring Promiscuity Space
The ‘vastness of sequence space’, i.e. the extremely large number of possible permutations in even the smallest catalytic proteins, requires evolutionary shortcuts. In this project the candidate will interconvert members of a prom...
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Descripción del proyecto
The ‘vastness of sequence space’, i.e. the extremely large number of possible permutations in even the smallest catalytic proteins, requires evolutionary shortcuts. In this project the candidate will interconvert members of a promiscuous catalytic superfamily into one anotehr and differentiate a multi-specific enzyme into mono-specific mutants. Ultimately it is hoped that directed evolution experiments will have implications for the definition of ‘phylogenetic relationships’ for catalysis. To this end the candidate will set up biochemical assays for hydrolytic reactions in small emulsion droplets (diameter: 2-20 um) and explore whether (i) these assays can be carried out in such in vitro compartemnts , (ii) whether it is possible to measure kinetics in these droplets oand (iii) carry out directed evolutino experiments with a sreening system of unprecedented capacity. In addition to tehse technical aspects, the candidate will also charcterise the mechanism of the wild type enzyme and its mutants.