# Directed Evolution of Halogenases for Small Molecule Functionalization

> **NIH NIH R01** · TRUSTEES OF INDIANA UNIVERSITY · 2020 · $317,000

## Abstract

Project Summary
 Halogenated organic compounds are used extensively as building blocks, synthetic intermediates, and end
use products for pharmaceutical and agrochemical applications. The utility of these compounds, including their
biological activity, arises from the reactivity and physical properties uniquely conferred to them by halogen
substitution. The importance of halogenation and limitations associated with current halogenation methods
prompted us to develop new halogenation catalysts. This proposal outlines the evolution of flavin dependent
halogenases (FDHs) and Fe(II) α-ketoglutarate dependent halogenases (FeDHs) for a range synthetic
methods involving selective halogenation and related (i.e. pseudohalogen) atom transfer reactions.
 Work completed in the concluding project period included the first first examples of FDH directed evolution
to improve the substrate scope, selectivity, and reactivity of these enzymes for practical synthetic applications.
Structural, kinetic, and computational methods have since been used to provide rigorous characterization of
the engineered enzymes and to inform subsequent engineering efforts. In the proposed project period, more
advanced synthetic applications of these enzymes, including halogenation of electron deficient substrates,
sequential halogenation/cross-coupling for lead diversification, and enantioselective halogenation will be
pursued. In a major new direction, FeDHs will also be evolved to catalyze non-directed site-selective sp3 C-H
halogenation, azidation, and other atom transfer reactions. These FDH and FeDH efforts will involve new
advances in genome mining and directed evolution methodology to expedite the identification and optimization
of enzymes for non-native catalysis, C-H bond functionalization, and synthetic applications.

## Key facts

- **NIH application ID:** 9993535
- **Project number:** 5R01GM115665-06
- **Recipient organization:** TRUSTEES OF INDIANA UNIVERSITY
- **Principal Investigator:** JARED C LEWIS
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $317,000
- **Award type:** 5
- **Project period:** 2015-08-15 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9993535

## Citation

> US National Institutes of Health, RePORTER application 9993535, Directed Evolution of Halogenases for Small Molecule Functionalization (5R01GM115665-06). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9993535. Licensed CC0.

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