Analysis of Laboratory-Evolved Flavin-Dependent Halogenases Affords a Computational Model for Predicting Halogenase Site Selectivity.
- Resource Type
- Academic Journal
- Authors
- Andorfer MC; Department of Chemistry, University of Chicago, Chicago, IL 60637, USA.; Present address: Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA.; These authors contributed equally.; Evans D; Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, CA 90095, USA.; These authors contributed equally.; Yang S; Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, CA 90095, USA.; Present address: Merck Research Laboratories, South San Francisco, CA, USA.; He CQ; Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, CA 90095, USA.; Present address: Merck Research Laboratories, Kenilworth, NJ, USA.; Girlich AM; Department of Chemistry, University of Chicago, Chicago, IL 60637, USA.; Vergara-Coll J; Department of Chemistry, University of Chicago, Chicago, IL 60637, USA.; Sukumar N; NE-CAT and Department of Chemistry and Chemical Biology, Cornell University, Argonne National Laboratory, Building 436E, Argonne, IL 60439, USA.; Houk KN; Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, CA 90095, USA.; Lewis JC; Department of Chemistry, Indiana University, Bloomington, IN 47405, USA.; Lead contact.
- Source
- Publisher: Elsevier B.V Country of Publication: Netherlands NLM ID: 101776366 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2667-1093 (Electronic) Linking ISSN: 26671093 NLM ISO Abbreviation: Chem Catal Subsets: PubMed not MEDLINE
- Subject
- Language
- English
Flavin-dependent halogenases (FDHs) catalyze selective halogenation of electron-rich aromatic compounds without the need for harsh oxidants required by conventional oxidative halogenation reactions. Predictive models for halogenase site selectivity could greatly improve their utility for chemical synthesis. Toward this end, we analyzed the structures and selectivity of three halogenase variants evolved to halogenate tryptamine with orthogonal selectivity. Crystal structures and reversion mutations revealed key residues involved in altering halogenase selectivity. Density functional theory calculations and molecular dynamics simulations are both consistent with hypohalous acid as the active halogenating species in FDH catalysis. This model was used to accurately predict the site selectivity of halogenase variants toward different synthetic substrates, providing a valuable tool for implementing halogenases in biocatalysis efforts.
Competing Interests: Declaration of Interests The authors declare no competing interests.