# De novo designed metalloproteins as a new generation of artificial hydrogenases

> **NIH NIH R15** · UNIVERSITY OF MISSISSIPPI · 2021 · $419,483

## Abstract

PROJECT SUMMARY/ABSTRACT
Hydrogenases are complex, metal-containing enzymes that generate energy for certain organisms by
catalyzing the reversible interconversion between H+ and H2 gas. Unraveling the intricate details about the
function of these enzymes will significantly advance the H2-based, carbon-neutral alternative energy
production. However, the complexity of these enzymes due to the presence of multiple metallic cofactors, low
production yield, and deactivation, makes studying these enzymes challenging. Our long term goals are to
design artificial biomolecular hydrogenases (ArHs) as simpler functional analogs of these metalloenzymes.
De novo metalloprotein design is an appealing and well-established approach to model complex
metalloproteins within minimal protein scaffolds. Although the designed systems are less complex, they serve
as water-soluble functional analogs of the native metalloenzymes and provide a functional view of the
chemistry. Employing this approach, we propose to pursue three Specific Aims describing the overall design
principles and functional/mechanistic attributes of the ArHs inspired by the [NiFe] hydrogenases. The overall
objectives of this proposal are: i) to design mononuclear (Ni), binuclear (Ni-Fe), and multinuclear (Ni3) active
sites within suitable de novo scaffolds; ii) characterize the physical and catalytic properties of the ArHs; iii)
determine the timescales of electron transfer; iv) outline the H+ transfer pathways; v) characterize the reaction
intermediates; and vi) elucidate how metals and protein scaffold work in synergy to influence the
properties/reactivity, such that a holistic mechanistic view of H-H bond formation can be attained. Our strong
preliminary results presented here attest that our objectives are achievable.
Collectively, the results from this proposed work will impact the fields of metalloprotein design, bioinorganic
chemistry, and alternative energy research. A novel class of ArHs will emerge, which will provide functional
vignettes into the working principles of H+ reduction pertaining to the native enzymes. The modular design
parameters and outcomes from this study will enable us to prepare biosynthetic catalysts with novel properties
and functions in the future.

## Key facts

- **NIH application ID:** 10360279
- **Project number:** 2R15GM131260-02
- **Recipient organization:** UNIVERSITY OF MISSISSIPPI
- **Principal Investigator:** Saumen Chakraborty
- **Activity code:** R15 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $419,483
- **Award type:** 2
- **Project period:** 2018-09-20 → 2025-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10360279

## Citation

> US National Institutes of Health, RePORTER application 10360279, De novo designed metalloproteins as a new generation of artificial hydrogenases (2R15GM131260-02). Retrieved via AI Analytics 2026-07-07 from https://api.ai-analytics.org/grant/nih/10360279. Licensed CC0.

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