Further Characterization of Fungal Halogenase RadH and Its Homologs

Further Characterization of Fungal Halogenase RadH and Its Homologs

Biomolecules

10.3390/biom13071081

http://dx.doi.org/10.3390/biom13071081

GuangRong Peh, Gregory A. Gunawan, Terence Tay, Elaine Tiong, Lee Ling Tan, Shimin Jiang, Yi Ling Goh, Suming Ye, Joel Wong, Christopher J. Brown, Huimin Zhao, Ee Lui Ang, Fong Tian Wong, Yee Hwee Lim

07 July 2023

Peh, G., Gunawan, G. A., Tay, T., Tiong, E., Tan, L. L., Jiang, S., Goh, Y. L., Ye, S., Wong, J., Brown, C. J., Zhao, H., Ang, E. L., Wong, F. T., & Lim, Y. H. (2023). Further Characterization of Fungal Halogenase RadH and Its Homologs. Biomolecules, 13(7), 1081. https://doi.org/10.3390/biom13071081

RadH is one of the flavin-dependent halogenases that has previously exhibited promising catalytic activity towards hydroxycoumarin, hydroxyisoquinoline, and phenolic derivatives. Here, we evaluated new functional homologs of RadH and expanded its specificities for the halogenation of non-tryptophan-derived, heterocyclic scaffolds. Our investigation revealed that RadH could effectively halogenate hydroxyquinoline and hydroxybenzothiophene. Assay optimization studies revealed the need to balance the various co-factor concentrations and where a GDHi co-factor recycling system most significantly improves the conversion and efficiency of the reaction. A crystal structure of RadH was also obtained with a resolution of 2.4 Å, and docking studies were conducted to pinpoint the binding and catalytic sites for substrates.

Attribution 4.0 International (CC BY 4.0)

This research / project is supported by the National Research Foundation (NRF) - Competitive Research Programme (CRP)
Grant Reference no. : NRF-CRP19-2017-05-00

This research is supported by core funding from: Agency for Science, Technology, and Research (A*STAR)
Grant Reference no. : C211917003

This research is supported by core funding from: Agency for Science, Technology, and Research (A*STAR)
Grant Reference no. : C211917004

This research is supported by core funding from: Agency for Science, Technology, and Research (A*STAR)
Grant Reference no. : C211917006

This research is supported by core funding from: Agency for Science, Technology, and Research (A*STAR)
Grant Reference no. : C211917010

A*STAR Graduate Academy (A*GA)

https://oar.a-star.edu.sg/communities-collections/articles/20479

2218-273X

Institute of Sustainability for Chemicals, Energy and Environment