# Novel Strategies in Light-driven P450 Enzymes

> **NIH NIH R16** · SAN JOSE STATE UNIVERSITY · 2022 · $146,500

## Abstract

Project Summary:
Cytochromes P450 are a superfamily of heme-thiolate monooxygenases, found in almost all living organisms,
that play an important role in the biosynthesis and biodegradation of endogenous compounds. In humans,
P450s are the major enzymes involved in drug metabolism and bioactivation, accounting for 75% of the total
metabolism. Harnessing the synthetic potential of these monooxygenases has also led to the recent use of
P450 enzymes in late-stage functionalization of various compounds. Our laboratory has developed an efficient
light-driven P450 hybrid enzyme system that utilizes the photochemical properties of Ru(II)-diimine
photosensitizers covalently attached to strategically positioned non-native single cysteine mutants of their
heme domain to perform P450 reactions upon visible light excitation. We recently started to investigate a
complementary bimolecular approach to activate P450 enzymes with peroxygenase activity, which is the focus
of Aim 1. Using various protein engineering approaches, we are assembling a large and diverse library of light-
initiated P450 enzymes with unique photocatalytic activity towards a range of substrates and with human
P450-like activities. We have also recently focused on developing chemoenzymatic approaches by marrying
the advantages of chemical catalysis with the selectivity of the P450 biocatalysis. We identified a promising
substrate anchoring approach to enhance the catalytic activity of P450 enzymes and alter their regioselectivity.
In Aim 2, we will capitalize on those strategies to synthesize a library of biphenylalkanoic acid and
diphenylamine derivatives, as potential dual fatty acid amide hydrolase (FAAH)/cyclooxygenase (COX)
inhibitors to mitigate gastric damages, currently plaguing the use of nonsteroidal anti-inflammatory drugs
(NSAIDs). A combination of computer docking simulation and biochemical assays will be used to ascertain
their inhibitory activity. In Aim 3, we will apply concepts identified in the substrate engineering approach to
develop a new platform for late stage substrate functionalization.
Our long-term goal is to employ the libraries of light-driven P450 enzymes combined with the chemoenzymatic
and substrate engineering strategies for the development of novel drugs and late stage diversification of lead
compounds.

## Key facts

- **NIH application ID:** 10410303
- **Project number:** 1R16GM145401-01
- **Recipient organization:** SAN JOSE STATE UNIVERSITY
- **Principal Investigator:** Lionel E. Cheruzel
- **Activity code:** R16 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $146,500
- **Award type:** 1
- **Project period:** 2022-05-05 → 2022-06-03

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10410303

## Citation

> US National Institutes of Health, RePORTER application 10410303, Novel Strategies in Light-driven P450 Enzymes (1R16GM145401-01). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10410303. Licensed CC0.

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