Design of efficient artificial enzymes using crystallographically-enhanced conformational sampling.

ABSTRACT: The ability to create efficient artificial enzymes for any chemical reaction is of great interest. Here, we describe a computational design method for increasing catalytic efficiency of de novo enzymes to a level comparable to their natural counterparts without relying on directed evolution. Using structural ensembles generated from dynamics-based refinement against X-ray diffraction data collected from crystals of Kemp eliminases HG3 (k_cat/K_M 125 M^-1 s^-1) and KE70 (k_cat/K_M 57 M^-1 s^-1), we design from each enzyme ≤10 sequences predicted to catalyze this reaction more efficiently. The most active designs display k_cat/K_M values improve

SUBMITTER: Rakotoharisoa RV 

PROVIDER: S-EPMC10635043 | biostudies-literature | 2023 Nov

REPOSITORIES: biostudies-literature