Expanding the Substrate Scope of Native Amine Dehydrogenases through In Silico Structural Exploration and Targeted Protein Engineering

Autor: Laurine Ducrot, Megan Bennett, Gwenaëlle André‐Leroux, Eddy Elisée, Sacha Marynberg, Aurélie Fossey‐Jouenne, Anne Zaparucha, Gideon Grogan, Carine Vergne‐Vaxelaire
Přispěvatelé: Génomique métabolique (UMR 8030), Genoscope - Centre national de séquençage [Evry] (GENOSCOPE), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), Structural Biology Laboratory, Department of Chemistry, University of York, Mathématiques et Informatique Appliquées du Génome à l'Environnement [Jouy-En-Josas] (MaIAGE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Region Ile-de-France, Diamond Light Source Didcot UKmx24948, ANR-19-CE07-0007,MODAMDH,Découverte et étude d'amine dehydrogénases par approche in silico(2019)
Rok vydání: 2022
Předmět:
Zdroj: ChemCatChem
ChemCatChem, 2022, 2022, pp.e202200880. ⟨10.1002/cctc.202200880⟩
ISSN: 1867-3899
1867-3880
DOI: 10.1002/cctc.202200880
Popis: International audience; Native Amine Dehydrogenases (nat-AmDHs) are NAD(P)H-enzymes performing reductive amination, mainly active towards small aliphatic aldehydes and cyclic ketones, due to active site volumes limited by the presence of several bulky amino acids. Herein, inspired by the diversity of residues at these positions among the family, we report the implementation of mutations F140A and W145A in CfusAmDH and their transposition in nine other members. Moderate to high conversions were obtained with substrates not accepted by the native enzymes, notably n-alkylaldehydes (44.6% - 99.5% for hexanal to nonanal) and n-alkylketones (16.0% - 53.7% for hexan-2-one to nonan-2-one) with retention of excellent (S)- enantioselectivity (>99% ee). Complementary to the reported (R)- selective AmDHs, the promising mutant CfusAmDH-W145A was further characterized for its synthetic potential. Crystal structure resolution and molecular dynamics gave insights into the cofactor and substrate specificity and the whole structural dynamics, thus providing keys for mutagenesis work on this enzyme family
Databáze: OpenAIRE
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