Computational design of novel enzymes without cofactors

In this review we present a recently developed computational method to design de novo enzymes. Starting from the three-dimensional arrangement of the transition state structure and the catalytic side chains around it (theozyme), RosettaMatch identifies successful placements of the theozyme into protein scaffolds. Subsequently, RosettaEnzDes (for EnzymeDesign) redesigns the active site around the theozyme for binding and stabilization of the transition state and the catalytic residues. The resulting computationally designed enzymes are expressed and experimentally tested for catalytic activity.