Further Characterization of Fungal Halogenase RadH and Its Homologs
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
Affiliations
GuangRong Peh
Institute of Sustainability for Chemicals, Energy and Environment (ISCE2), Agency for Science, Technology and Research (A*STAR), 8 Biomedical Grove, Neuros #07-01, Singapore 138665, Singapore
Gregory A. Gunawan
Institute of Sustainability for Chemicals, Energy and Environment (ISCE2), Agency for Science, Technology and Research (A*STAR), 8 Biomedical Grove, Neuros #07-01, Singapore 138665, Singapore
Terence Tay
Singapore Institute of Food and Biotechnology Innovation (SIFBI), Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos #01-02, Singapore 138669, Singapore
Elaine Tiong
Institute of Molecular and Cell Biology (IMCB), Agency for Science, Technology and Research (A*STAR), 61 Biopolis Dr, Proteos #07-01, Singapore 138673, Singapore
Lee Ling Tan
Institute of Molecular and Cell Biology (IMCB), Agency for Science, Technology and Research (A*STAR), 61 Biopolis Dr, Proteos #07-01, Singapore 138673, Singapore
Shimin Jiang
Disease Intervention Technology Laboratory, Institute of Molecular and Cellular Biology, Agency for Science, Technology and Research (A*STAR), 8A Biomedical Grove, Neuros/Immunos #06-04/05, Singapore 138648, Singapore
Yi Ling Goh
Institute of Sustainability for Chemicals, Energy and Environment (ISCE2), Agency for Science, Technology and Research (A*STAR), 8 Biomedical Grove, Neuros #07-01, Singapore 138665, Singapore
Suming Ye
Institute of Sustainability for Chemicals, Energy and Environment (ISCE2), Agency for Science, Technology and Research (A*STAR), 8 Biomedical Grove, Neuros #07-01, Singapore 138665, Singapore
Joel Wong
Institute of Sustainability for Chemicals, Energy and Environment (ISCE2), Agency for Science, Technology and Research (A*STAR), 8 Biomedical Grove, Neuros #07-01, Singapore 138665, Singapore
Christopher J. Brown
Disease Intervention Technology Laboratory, Institute of Molecular and Cellular Biology, Agency for Science, Technology and Research (A*STAR), 8A Biomedical Grove, Neuros/Immunos #06-04/05, Singapore 138648, Singapore
Huimin Zhao
Singapore Institute of Food and Biotechnology Innovation (SIFBI), Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos #01-02, Singapore 138669, Singapore
Ee Lui Ang
Singapore Institute of Food and Biotechnology Innovation (SIFBI), Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos #01-02, Singapore 138669, Singapore
Fong Tian Wong
Institute of Sustainability for Chemicals, Energy and Environment (ISCE2), Agency for Science, Technology and Research (A*STAR), 8 Biomedical Grove, Neuros #07-01, Singapore 138665, Singapore
Yee Hwee Lim
Institute of Sustainability for Chemicals, Energy and Environment (ISCE2), Agency for Science, Technology and Research (A*STAR), 8 Biomedical Grove, Neuros #07-01, Singapore 138665, Singapore
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.
