Please use this identifier to cite or link to this item: https://doi.org/10.21256/zhaw-22632
Publication type: Article in scientific journal
Type of review: Peer review (publication)
Title: Re‐programming and optimization of a L‐proline cis‐4‐hydroxylase for the cis‐3‐halogenation of its native substrate
Authors: Papadopoulou, Athena
Meierhofer, Jasmin
Meyer, Fabian
Hayashi, Takahiro
Schneider, Samuel
Sager, Emine
Buller, Rebecca
et. al: No
DOI: 10.1002/cctc.202100591
10.21256/zhaw-22632
Published in: ChemCatChem
Volume(Issue): 13
Issue: 18
Page(s): 3914
Pages to: 3919
Issue Date: 7-Jun-2021
Publisher / Ed. Institution: Wiley
ISSN: 1867-3880
1867-3899
Language: English
Subject (DDC): 660.6: Biotechnology
Abstract: Non-heme iron/ α -ketoglutarate dependent halogenases acting on freestanding substrates catalyze the regio- and stereoselective halogenation of inactivated C(sp 3 )-H bonds. Yet, with only a handful of these halogenases characterized, the biosynthetic potential of enzymatic radical halogenation remains limited. Herein, we describe the remodeling of L -proline cis -4-hydroxylase from Sinorhizobium meliloti into a halogenase by introduction of a single point mutation ( D108G) into the enzyme’s active site. The re-programmed halogenase displays a striking regio-divergent reaction chemistry: While halogenation of L -proline exclusively occurs at the C3-position, the retained hydroxylation activity leads to derivatization at the C-4 position, corresponding to the regioselectivity of the wildtype enzyme. By employing several rounds of directed evolution, an optimized halogenase variant with 98-fold improved apparent k cat / K m for chlorination of L -proline compared to the parental enzyme SmP4H ( D108G) was identified. The development and optimization of this novel halogenation biocatalyst highlights the possibility to rationally harness the chemical versatility of non-heme Fe/ α KG dependent dioxygenases for C-H functionalization .
URI: https://digitalcollection.zhaw.ch/handle/11475/22632
Fulltext version: Published version
License (according to publishing contract): CC BY 4.0: Attribution 4.0 International
Restricted until: 2022-06-08
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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