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|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|
|Publisher / Ed. Institution:||Wiley|
|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 .|
|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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|2021_Papadopoulou-etal_Re-programming-and-optimization_VoR.pdf||Published Version||1.84 MB||Adobe PDF|
|2021_Papadopoulou-etal_Re-programming-and-optimization.pdf||Accepted Version||885.69 kB||Adobe PDF|
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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, 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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