Modulating chemoselectivity in a Fe(II)/α-ketoglutarate-dependent dioxygenase for the oxidative modification of a non-proteinogenic amino acid

Meyer, Fabian; Frey, Raphael; Ligibel, Mathieu; Sager, Emine; Schroer, Kirsten; Snajdrova, Radka; Buller, Rebecca

doi:10.1021/acscatal.1c00678

Please use this identifier to cite or link to this item: https://doi.org/10.21256/zhaw-22606

Publication type:	Article in scientific journal
Type of review:	Peer review (publication)
Title:	Modulating chemoselectivity in a Fe(II)/α-ketoglutarate-dependent dioxygenase for the oxidative modification of a non-proteinogenic amino acid
Authors:	Meyer, Fabian Frey, Raphael Ligibel, Mathieu Sager, Emine Schroer, Kirsten Snajdrova, Radka Buller, Rebecca
et. al:	No
DOI:	10.1021/acscatal.1c00678 10.21256/zhaw-22606
Published in:	ACS Catalysis
Volume(Issue):	2021
Issue:	11
Issue Date:	10-May-2021
Publisher / Ed. Institution:	American Chemical Society
ISSN:	2155-5435
Language:	English
Subject (DDC):	660.6: Biotechnology
Abstract:	Modification of aliphatic C–H bonds in a regio- and stereoselective manner can pose a formidable challenge in organic chemistry. In this context, the use of nonheme iron and α-ketoglutarate-dependent dioxygenases (αKGDs) represents an interesting tool for C–H activation as this enzyme family can catalyze a broad set of synthetically valuable reactions including hydroxylations, oxidations, and desaturations. The consensus reaction mechanism of αKGDs proceeds via the formation of a Fe(IV)-oxo complex capable of hydrogen atom transfer (HAT) from an sp3-hybridized substrate carbon center. The resulting substrate radical and Fe(III)–OH cofactor are considered to be the branch point toward the possible reaction outcomes which are determined by the enzyme’s active site architecture. To date, the modulation of the reaction fate in Fe(II)/α-ketoglutarate-dependent dioxygenases via enzyme engineering has been mainly elusive. In this study, we therefore set out to engineer the l-proline cis-4-hydroxylase SmP4H from Sinorhizobium meliloti for selective oxidative modifications of the nonproteinogenic amino acid l-homophenylalanine (l-hPhe) to produce pharmacological relevant small molecule intermediates. Using structure-guided directed evolution, we improved the total turnover number, the kcat, as well as the kcat/Km of the hydroxylation reaction yielding the desired γ-hydroxylation product by approximately 10-fold, >100-fold, and >300-fold, respectively. Notably, the exchange of only one amino acid in the active site (W40Y) allowed us to reprogram the natural hydroxylase to predominantly act as a desaturase, presumably through tyrosine’s capability to serve as a catalytic entity in the reaction mechanism. An investigation of the substrate scope revealed additional acceptance of the noncanonical amino acids l-homotyrosine and (S)-α-amino-3,4-dichlorobenzenebutanoic acid by SmP4H variants.
URI:	https://digitalcollection.zhaw.ch/handle/11475/22606
Fulltext version:	Accepted version
License (according to publishing contract):	Licence according to publishing contract
Restricted until:	2022-05-10
Departement:	Life Sciences and Facility Management
Organisational Unit:	Institute of Chemistry and Biotechnology (ICBT)
Appears in collections:	Publikationen Life Sciences und Facility Management

Files in This Item:

File	Description	Size	Format
Manuscript_Modulating chemoselectivity in a Fe(II) α-ketoglutarate dependent dioxygenase for the oxidative modification of a non-proteinogenic amino.pdf	Accepted Article	1.73 MB	Adobe PDF	View/Open
SupI_Modulating chemoselectivity in a Fe(II) α-ketoglutarate dependent dioxygenase for the oxidative modification of a non-proteinogenic amino acid.pdf	Supporting Information	2.59 MB	Adobe PDF	View/Open

Show full item record

Meyer, F., Frey, R., Ligibel, M., Sager, E., Schroer, K., Snajdrova, R., & Buller, R. (2021). Modulating chemoselectivity in a Fe(II)/α-ketoglutarate-dependent dioxygenase for the oxidative modification of a non-proteinogenic amino acid. ACS Catalysis, 2021(11). https://doi.org/10.1021/acscatal.1c00678

Meyer, F. et al. (2021) ‘Modulating chemoselectivity in a Fe(II)/α-ketoglutarate-dependent dioxygenase for the oxidative modification of a non-proteinogenic amino acid’, ACS Catalysis, 2021(11). Available at: https://doi.org/10.1021/acscatal.1c00678.

F. Meyer et al., “Modulating chemoselectivity in a Fe(II)/α-ketoglutarate-dependent dioxygenase for the oxidative modification of a non-proteinogenic amino acid,” ACS Catalysis, vol. 2021, no. 11, May 2021, doi: 10.1021/acscatal.1c00678.

MEYER, Fabian, Raphael FREY, Mathieu LIGIBEL, Emine SAGER, Kirsten SCHROER, Radka SNAJDROVA und Rebecca BULLER, 2021. Modulating chemoselectivity in a Fe(II)/α-ketoglutarate-dependent dioxygenase for the oxidative modification of a non-proteinogenic amino acid. ACS Catalysis. 10 Mai 2021. Bd. 2021, Nr. 11. DOI 10.1021/acscatal.1c00678

Meyer, Fabian, Raphael Frey, Mathieu Ligibel, Emine Sager, Kirsten Schroer, Radka Snajdrova, and Rebecca Buller. 2021. “Modulating Chemoselectivity in a Fe(II)/α-Ketoglutarate-Dependent Dioxygenase for the Oxidative Modification of a Non-Proteinogenic Amino Acid.” ACS Catalysis 2021 (11). https://doi.org/10.1021/acscatal.1c00678.

Meyer, Fabian, et al. “Modulating Chemoselectivity in a Fe(II)/α-Ketoglutarate-Dependent Dioxygenase for the Oxidative Modification of a Non-Proteinogenic Amino Acid.” ACS Catalysis, vol. 2021, no. 11, May 2021, https://doi.org/10.1021/acscatal.1c00678.