Design of an allosterically regulated retroaldolase

• DOI: 10.1002/pro.2622
• Publication Date: 2014-12-16T21:21:46Z
• Authors: Elizabeth A. Raymond; Korrie L. Mack; Jennifer H. Yoon; Olesia V. Moroz; Yurii S. Moroz; Ivan V. Korendovych
• Keywords: Residue (chemistry)

We employed a minimalist approach for design of an allosterically controlled retroaldolase. Introduction single lysine residue into the nonenzymatic protein calmodulin led to 15,000-fold increase in second order rate constant retroaldol reaction with methodol as substrate. The resulting catalyst AlleyCatR is active enough subsequent directed evolution crude cell bacterial lysates. AlleyCatR's activity regulated by Ca(2+) ions. No catalysis observed absence metal ion. catalytic originates from hydrophobic interaction substrate (∼2000-fold) and change apparent pKa residue.