A single mutation in a regulatory protein produces evolvable allosterically regulated catalyst of nonnatural reaction

Abstract

It only takes one mutation: a strategically placed single mutation in a non-enzymatic protein scaffold produced AlleyCat, a small, allosterically regulated catalyst of Kemp elimination. In only 7 rounds of directed evolution enzymatic efficiency of the original 74 amino acid residue catalyst was improved more than 220-fold to achieve k_cat value higher than that of catalytic antibodies for the same reaction, still preserving allosteric regulation.