Targeted Enzyme Engineering Unveiled Unexpected Patterns of Halogenase Stabilization

Minges H, Schnepel C, Boettcher D, Weiss MS, Sproß J, Bornscheuer UT, Sewald N (2020)
ChemCatChem 12(3): 818-831.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Autor*in
Minges, HannahUniBi; Schnepel, ChristianUniBi; Boettcher, Dominique; Weiss, Martin S.; Sproß, JensUniBi; Bornscheuer, Uwe T.; Sewald, NorbertUniBi
Abstract / Bemerkung
Halogenases are valuable biocatalysts for selective C-H activation, but despite recent efforts to broaden their application scope by means of protein engineering, improvement of thermostability and catalytic efficiency is still desired. A directed evolution campaign aimed at generating a thermostable flavin-dependent tryptophan 6-halogenase with reasonable activity suitable for chemoenzymatic purposes. These characteristics were tackled by combining successive rounds of epPCR along with semi-rational mutagenesis leading to a triple mutant (Thal-GLV) with substantially increased thermostability (T-M=23.5 K) and higher activity at 25 degrees C than the wild type enzyme. Moreover, an active-site mutation has a striking impact on thermostability but also on enantioselectivity. Our data contribute to a detailed understanding of biohalogenation and provide a profound basis for future engineering strategies to facilitate chemoenzymatic application of these attractive biocatalysts.
Stichworte
directed evolution; enzyme catalysis; enzyme stability; rational; mutagenesis; tryptophan halogenase
Erscheinungsjahr
2020
Zeitschriftentitel
ChemCatChem
Band
12
Ausgabe
3
Seite(n)
818-831
ISSN
1867-3880
eISSN
1867-3899
Page URI
https://pub.uni-bielefeld.de/record/2939933

Zitieren

Minges H, Schnepel C, Boettcher D, et al. Targeted Enzyme Engineering Unveiled Unexpected Patterns of Halogenase Stabilization. ChemCatChem. 2020;12(3):818-831.
Minges, H., Schnepel, C., Boettcher, D., Weiss, M. S., Sproß, J., Bornscheuer, U. T., & Sewald, N. (2020). Targeted Enzyme Engineering Unveiled Unexpected Patterns of Halogenase Stabilization. ChemCatChem, 12(3), 818-831. doi:10.1002/cctc.201901827
Minges, H., Schnepel, C., Boettcher, D., Weiss, M. S., Sproß, J., Bornscheuer, U. T., and Sewald, N. (2020). Targeted Enzyme Engineering Unveiled Unexpected Patterns of Halogenase Stabilization. ChemCatChem 12, 818-831.
Minges, H., et al., 2020. Targeted Enzyme Engineering Unveiled Unexpected Patterns of Halogenase Stabilization. ChemCatChem, 12(3), p 818-831.
H. Minges, et al., “Targeted Enzyme Engineering Unveiled Unexpected Patterns of Halogenase Stabilization”, ChemCatChem, vol. 12, 2020, pp. 818-831.
Minges, H., Schnepel, C., Boettcher, D., Weiss, M.S., Sproß, J., Bornscheuer, U.T., Sewald, N.: Targeted Enzyme Engineering Unveiled Unexpected Patterns of Halogenase Stabilization. ChemCatChem. 12, 818-831 (2020).
Minges, Hannah, Schnepel, Christian, Boettcher, Dominique, Weiss, Martin S., Sproß, Jens, Bornscheuer, Uwe T., and Sewald, Norbert. “Targeted Enzyme Engineering Unveiled Unexpected Patterns of Halogenase Stabilization”. ChemCatChem 12.3 (2020): 818-831.
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