Engineering Tryptophan Halogenase RebH for Regioselective Halogenation of Phenolic Compounds
Herein, we engineered tryptophan halogenase RebH using alanine scanning and the focused rational iterative site-specific mutagenesis (FRISM) strategy to expand its substrate scope toward phenolic compounds, particularly hydroxyquinolines and hydroxyisoquinolines, with excellent regioselectivity. The optimal mutant, 3-LSR/V80A/F111G, exhibited a 5-fold increase in catalytic activity and an 86.5-fold enhancement in catalytic efficiency (<i>k</i><sub>cat</sub>/<i>K</i><sub>m</sub>) for 6-hydroxyisoquinoline compared with the template 3-LSR. Furthermore, 3-LSR/V80A/F111G enabled the synthesis of 5-bromo-6-hydroxyisoquinoline on a milligram-scale with >99.9% conversion and 84.3% isolated yield.
Authors
- Wenhe Zhang
- Qizhen Wang
- Yutian Jia
- Xian Jia (ORCID: https://orcid.org/0000-0003-4311-2742)
- Song You (ORCID: https://orcid.org/0000-0002-6553-6602)
- Jiuming Zhang (ORCID: https://orcid.org/0000-0003-1129-1870)
- Jianxin Li (ORCID: https://orcid.org/0000-0002-9059-330X)
- Bin Qin (ORCID: https://orcid.org/0000-0002-9180-0550)
- Jingbo Wanchao
Institutions
- Shenyang Pharmaceutical University (CN)
- The University of Tokyo (JP)
- Kumamoto University (JP)
Publication Details
- Journal
- Organic Letters
- Published
- 2025-09-18
- DOI
- https://doi.org/10.1021/acs.orglett.5c03448
- Primary Topic
- Vanadium and Halogenation Chemistry
- Type
- article
- Field-Weighted Citation Impact
- 0.00