Work

High-Throughput Screening of Co-Protoporphyrin IX-Binding Proteins for Enhanced Hydrogen Production

Year: 2026

Type: article

Abstract: Artificial metalloenzymes incorporating cobalt protoporphyrin IX (Co-PPIX) are promising for sustainable hydrogen production; however, slow protein preparation and a lack of suitable detection methods limit the systematic optimization of their catalytic performance. Here, we report a streamlined workflow that combines the direct in vivo incorporation of Co-PPIX into cytochrome b562 (cyt b562) variants with a colorimetric assay for hydrogen evolution, scalable to hundreds of mutants. We screened 103 members of a mutant library and selected the variant Co-Mut25, which displayed activity double than wild type on the screen, and produced over 70% more hydrogen than WT as assessed by gas chromatography. This approach enables the rapid and scalable identification of high-performing cobalt–protein catalysts and expands the toolkit for artificial hydrogenase development.

Source: Molecules

DOI: 10.3390/molecules31020346

Authors: Nicholas Ryan Halloran, Mohammad Imtiazur Rahman, Roman Christopher Fabry, Abesh Banerjee, Giovanna Ghirlanda

Institution: Arizona State University

Language: English

Topic: Metalloenzymes and iron-sulfur proteins

Subfield: Renewable Energy, Sustainability and the Environment

Field: Energy

Domain: Physical Sciences

Open Access status: gold

Estimated APC paid: $3,258

Funder: National Science Foundation