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

Gideon James Grogan; Laurine  Ducrot; Megan Bennett; Gwenaelle Andre_Leroux; Eddy Elisee; Sacha Marynberg; Aurelie Fossey-Jouenne; Anne Zaparucha; Carine Vergne-Vaxelaire

doi:10.1002/cctc.202200880

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

Gideon James Grogan, Laurine Ducrot, Megan Bennett, Gwenaelle Andre_Leroux, Eddy Elisee, Sacha Marynberg, Aurelie Fossey-Jouenne, Anne Zaparucha, Carine Vergne-Vaxelaire

Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

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 Cfus AmDH 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 Cfus AmDH-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.

Original language	English
Journal	ChemCatChem
Early online date	29 Sept 2022
DOIs	https://doi.org/10.1002/cctc.202200880
Publication status	E-pub ahead of print - 29 Sept 2022

Bibliographical note

Access to Document

10.1002/cctc.202200880Licence: CC BY

Expandingthe SubstrateScopeof NativeAmineDehydrogenasesthroughIn SilicoStructuralExplorationand TargetedProteinEngineering
© 2022The Authors.
Final published version, 1.88 MBLicence: CC BY-NC-ND

Cite this

@article{d8d450c1c8824f94b57fcdec114f359c,

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

abstract = "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 Cfus AmDH 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 Cfus AmDH-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.",

author = "Grogan, {Gideon James} and Laurine Ducrot and Megan Bennett and Gwenaelle Andre_Leroux and Eddy Elisee and Sacha Marynberg and Aurelie Fossey-Jouenne and Anne Zaparucha and Carine Vergne-Vaxelaire",

note = "{\textcopyright} 2022The Authors.",

year = "2022",

month = sep,

day = "29",

doi = "10.1002/cctc.202200880",

language = "English",

journal = "ChemCatChem",

issn = "1867-3880",

publisher = "Wiley-Blackwell",

}

TY - JOUR

T1 - Expanding the Substrate Scope of Native Amine Dehydrogenases through In Silico Structural Exploration and Targeted Protein Engineering

AU - Grogan, Gideon James

AU - Ducrot, Laurine

AU - Bennett, Megan

AU - Andre_Leroux, Gwenaelle

AU - Elisee, Eddy

AU - Marynberg, Sacha

AU - Fossey-Jouenne, Aurelie

AU - Zaparucha, Anne

AU - Vergne-Vaxelaire, Carine

PY - 2022/9/29

Y1 - 2022/9/29

N2 - 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 Cfus AmDH 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 Cfus AmDH-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.

AB - 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 Cfus AmDH 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 Cfus AmDH-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.

U2 - 10.1002/cctc.202200880

DO - 10.1002/cctc.202200880

M3 - Article

SN - 1867-3880

JO - ChemCatChem

JF - ChemCatChem

ER -