Enzyme engineering enables inversion of substrate stereopreference of the halogenase WelO5*

Voss, Moritz; Hüppi, Sean; Schaub, Daniela; Hayashi, Takahiro; Ligibel, Mathieu; Sager, Emine; Schroer, Kirsten; Snajdrova, Radka; Buller, Rebecca

doi:10.1002/cctc.202201115

Please use this identifier to cite or link to this item: https://doi.org/10.21256/zhaw-26366

Publication type:	Article in scientific journal
Type of review:	Peer review (publication)
Title:	Enzyme engineering enables inversion of substrate stereopreference of the halogenase WelO5*
Authors:	Voss, Moritz Hüppi, Sean Schaub, Daniela Hayashi, Takahiro Ligibel, Mathieu Sager, Emine Schroer, Kirsten Snajdrova, Radka Buller, Rebecca
et. al:	No
DOI:	10.1002/cctc.202201115 10.21256/zhaw-26366
Published in:	ChemCatChem
Volume(Issue):	14
Issue:	24
Page(s):	e202201115
Issue Date:	2022
Publisher / Ed. Institution:	Wiley
ISSN:	1867-3880 1867-3899
Language:	English
Subjects:	Biocatalysis; Enzyme engineering
Subject (DDC):	660.6: Biotechnology
Abstract:	Enzymatic late-stage diversification of small molecules has the potential to rapidly generate diversity in compound libraries dedicated to drug discovery. In this context, freestanding Fe(II)/α-ketoglutarate-dependent halogenases have raised particular interest as this enzyme family allows the otherwise difficult regio- and stereoselective halogenation of unactivated C(sp3)-H bonds. Here, we report the development of two engineered variants of the halogenase WelO5* for the racemic resolution of a mixture of stereoisomers generated in the synthesis of a bioactive martinelline-derived fragment. By screening a 3-site combinatorial variant library, we could identify two variants exhibiting exquisite substrate selectivity towards the desired enantiomers. Strikingly, the inversion of substrate stereopreference between the halogenase variants was achieved by varying only three residues in the active site. Protein crystallization and subsequent structure elucidation of the wildtype enzyme and a WelO5* variant shed light on the factors governing substrate acceptance and selectivity.
URI:	https://digitalcollection.zhaw.ch/handle/11475/26366
Fulltext version:	Published version
License (according to publishing contract):	CC BY 4.0: Attribution 4.0 International
Departement:	Life Sciences and Facility Management
Organisational Unit:	Institute of Chemistry and Biotechnology (ICBT)
Appears in collections:	Publikationen Life Sciences und Facility Management

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Voss, M., Hüppi, S., Schaub, D., Hayashi, T., Ligibel, M., Sager, E., Schroer, K., Snajdrova, R., & Buller, R. (2022). Enzyme engineering enables inversion of substrate stereopreference of the halogenase WelO5*. ChemCatChem, 14(24), e202201115. https://doi.org/10.1002/cctc.202201115

Voss, M. et al. (2022) ‘Enzyme engineering enables inversion of substrate stereopreference of the halogenase WelO5*’, ChemCatChem, 14(24), p. e202201115. Available at: https://doi.org/10.1002/cctc.202201115.

M. Voss et al., “Enzyme engineering enables inversion of substrate stereopreference of the halogenase WelO5*,” ChemCatChem, vol. 14, no. 24, p. e202201115, 2022, doi: 10.1002/cctc.202201115.

VOSS, Moritz, Sean HÜPPI, Daniela SCHAUB, Takahiro HAYASHI, Mathieu LIGIBEL, Emine SAGER, Kirsten SCHROER, Radka SNAJDROVA und Rebecca BULLER, 2022. Enzyme engineering enables inversion of substrate stereopreference of the halogenase WelO5*. ChemCatChem. 2022. Bd. 14, Nr. 24, S. e202201115. DOI 10.1002/cctc.202201115

Voss, Moritz, Sean Hüppi, Daniela Schaub, Takahiro Hayashi, Mathieu Ligibel, Emine Sager, Kirsten Schroer, Radka Snajdrova, and Rebecca Buller. 2022. “Enzyme Engineering Enables Inversion of Substrate Stereopreference of the Halogenase WelO5*.” ChemCatChem 14 (24): e202201115. https://doi.org/10.1002/cctc.202201115.

Voss, Moritz, et al. “Enzyme Engineering Enables Inversion of Substrate Stereopreference of the Halogenase WelO5*.” ChemCatChem, vol. 14, no. 24, 2022, p. e202201115, https://doi.org/10.1002/cctc.202201115.