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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Papadopoulou, A., Meierhofer, J., Meyer, F., Hayashi, T., Schneider, S., Sager, E., & Buller, R. (2021). Re‐programming and optimization of a L‐proline cis‐4‐hydroxylase for the cis‐3‐halogenation of its native substrate. ChemCatChem, 13(18), 3914–3919. https://doi.org/10.1002/cctc.202100591
Papadopoulou, A. et al. (2021) ‘Re‐programming and optimization of a L‐proline cis‐4‐hydroxylase for the cis‐3‐halogenation of its native substrate’, ChemCatChem, 13(18), pp. 3914–3919. Available at: https://doi.org/10.1002/cctc.202100591.
A. Papadopoulou et al., “Re‐programming and optimization of a L‐proline cis‐4‐hydroxylase for the cis‐3‐halogenation of its native substrate,” ChemCatChem, vol. 13, no. 18, pp. 3914–3919, Jun. 2021, doi: 10.1002/cctc.202100591.
PAPADOPOULOU, Athena, Jasmin MEIERHOFER, Fabian MEYER, Takahiro HAYASHI, Samuel SCHNEIDER, Emine SAGER und Rebecca BULLER, 2021. Re‐programming and optimization of a L‐proline cis‐4‐hydroxylase for the cis‐3‐halogenation of its native substrate. ChemCatChem. 7 Juni 2021. Bd. 13, Nr. 18, S. 3914–3919. DOI 10.1002/cctc.202100591
Papadopoulou, Athena, Jasmin Meierhofer, Fabian Meyer, Takahiro Hayashi, Samuel Schneider, Emine Sager, and Rebecca Buller. 2021. “Re‐Programming and Optimization of a L‐Proline Cis‐4‐Hydroxylase for the Cis‐3‐Halogenation of Its Native Substrate.” ChemCatChem 13 (18): 3914–19. https://doi.org/10.1002/cctc.202100591.
Papadopoulou, Athena, et al. “Re‐Programming and Optimization of a L‐Proline Cis‐4‐Hydroxylase for the Cis‐3‐Halogenation of Its Native Substrate.” ChemCatChem, vol. 13, no. 18, June 2021, pp. 3914–19, https://doi.org/10.1002/cctc.202100591.


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