# Abiological enzymatic C-H functionalization for bioactive molecule construction and diversification

> **NIH NIH R01** · CALIFORNIA INSTITUTE OF TECHNOLOGY · 2021 · $68,884

## Abstract

PROJECT SUMMARY/ABSTRACT
Antibiotics are antibacterial drugs used to treat bacterial infections by killing the bacteria or inhibiting bacterial
growth to allow immune clearance. While antibiotics help treat infections, their use puts selective pressure on
bacteria and inevitably leads to the emergence of resistant bacteria. Additionally, the development of novel
antibiotics has slowed in recent years which has left us with fewer treatment options for multidrug-resistant
bacterial infections. Moreover, most of these problematic multidrug-resistant bacteria are gram-negative,
meaning they are double-membraned and thus intrinsically resistant to many drugs, making drug development
even more challenging. There is a dire need for innovative approaches to develop new antibiotics that are
effective against gram-negative bacteria so we can preserve our ability to treat these infections. Promising
studies have shown that the addition of primary amines can improve drug accumulation into gram-negative
bacteria for various compounds, making it a rational and feasible approach to increase the activity of promising
antibacterials by improving their bacterial permeation. Synthetically aminating bioactive compounds can be
onerous, typically requiring multiple steps and the addition of other functional groups before acquiring the desired
amine. In contrast, directed evolution of cytochromes P450 is a promising strategy to develop enzymatic
platforms for the late-stage modifications of bioactive compounds. The ability of P450 enzymes to functionalize
the inert yet ubiquitous C–H bonds in bioactive compounds can be exploited to facilitate the diversification of
natural products. Recent successes have expanded the natural activity of P450s to include primary amination of
benzylic and allylic C(sp3) – H bonds via C–H nitrene insertion. Here, I propose to expand further this primary
amination activity to include substrates that are natural antibacterials. Our approach to engineering enzymes to
directly aminate C–H bonds will provide a means to accelerate the amination of antibacterials. Streamlining the
amination process of antibacterials will in turn facilitate downstream studies so the antibacterial potential of these
aminated derivatives can be thoroughly investigated. These methods will help establish engineered aminases
for antibacterials as a proof of concept that can be expanded to help potentiate various antibacterials.

## Key facts

- **NIH application ID:** 10397244
- **Project number:** 3R01GM138740-02S1
- **Recipient organization:** CALIFORNIA INSTITUTE OF TECHNOLOGY
- **Principal Investigator:** FRANCES H ARNOLD
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $68,884
- **Award type:** 3
- **Project period:** 2020-07-01 → 2024-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10397244

## Citation

> US National Institutes of Health, RePORTER application 10397244, Abiological enzymatic C-H functionalization for bioactive molecule construction and diversification (3R01GM138740-02S1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10397244. Licensed CC0.

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