A Single Mutation in a Regulatory Protein Produces Evolvable Allosterically Regulated Catalyst of Nonnatural Reaction (pages 6246–6249)
Dr. Olesia V. Moroz, Dr. Yurii S. Moroz, Dr. Yibing Wu, Alissa B. Olsen, Dr. Hong Cheng, Korrie L. Mack, Jaclyn M. McLaughlin, Elizabeth A. Raymond, Krystyna Zhezherya, Prof. Dr. Heinrich Roder and Prof. Dr. Ivan V. Korendovych
Article first published online: 29 APR 2013 | DOI: 10.1002/anie.201302339
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 seven rounds of directed evolution the enzymatic efficiency of the original 74 amino acid residue catalyst was improved more than 220-fold to achieve a kcat value higher than that of catalytic antibodies for the same reaction, still preserving allosteric regulation.