Design of an enantioselective artificial metallo-hydratase enzyme containing an unnatural metal-binding amino acid

Ivana Drienovska, Lur Alonso-Cotchico, Pietro Vidossich, Agusti Lledos, Jean-Didier Marechal, Gerard Roelfes

OnderzoeksoutputAcademicpeer review

45 Citaten (Scopus)
244 Downloads (Pure)


The design of artificial metalloenzymes is a challenging, yet ultimately highly rewarding objective because of the potential for accessing new-to-nature reactions. One of the main challenges is identifying catalytically active substrate-metal cofactor-host geometries. The advent of expanded genetic code methods for the in vivo incorporation of non-canonical metal-binding amino acids into proteins allow to address an important aspect of this challenge: the creation of a stable, well-defined metal-binding site. Here, we report a designed artificial metallohydratase, based on the transcriptional repressor lactococcal multidrug resistance regulator (LmrR), in which the non-canonical amino acid (2,2'-bipyridin-5yl) alanine is used to bind the catalytic Cu(II) ion. Starting from a set of empirical pre-conditions, a combination of cluster model calculations (QM), protein-ligand docking and molecular dynamics simulations was used to propose metallohydratase variants, that were experimentally verified. The agreement observed between the computationally predicted and experimentally observed catalysis results demonstrates the power of the artificial metalloenzyme design approach presented here.

Originele taal-2English
Pagina's (van-tot)7228-7235
Aantal pagina's8
TijdschriftChemical Science
Nummer van het tijdschrift10
StatusPublished - okt-2017

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