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Oxygen-resistant [FeFe]hydrogenases: new biocatalysis tools for clean energy and cascade reactions

Valetti, Francesca; Morra, Simone; Barbieri, Lisa; Dezzani, Sabrina; Ratto, Alessandro; Catucci, Gianluca; Sadeghi, Sheila J.; Gilardi, Gianfranco


Francesca Valetti

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Assistant Professor in Chemical &environmental Engineering

Lisa Barbieri

Sabrina Dezzani

Alessandro Ratto

Gianluca Catucci

Sheila J. Sadeghi

Gianfranco Gilardi


The use of enzymes to generate hydrogen, instead of using rare metal catalysts, is an exciting area of study in modern biochemistry and biotechnology, as well as biocatalysis driven by sustainable hydrogen. Thus far, the oxygen sensitivity of the fastest hydrogen-producing/exploiting enzymes, [FeFe]hydrogenases, has hindered their practical application, thereby restricting innovations mainly to their [NiFe]-based, albeit slower, counterparts. Recent exploration of the biodiversity of clostridial hydrogen-producing enzymes has yielded the isolation of representatives from a relatively understudied group. These enzymes possess an inherent defense mechanism against oxygen-induced damage. This discovery unveils fresh opportunities for applications such as electrode interfacing, biofuel cells, immobilization, and entrapment for enhanced stability in practical uses. Furthermore, it suggests potential combinations with cascade reactions for CO2 conversion or cofactor regeneration, like NADPH, facilitating product separation in biotechnological processes. This work provides an overview of this new class of biocatalysts, incorporating unpublished protein engineering strategies to further investigate the dynamic mechanism of oxygen protection and to address crucial details remaining elusive such as still unidentified switching hot-spots and their effects. Variants with improved kcat as well as chimeric versions with promising features to attain gain-of-function variants and applications in various biotechnological processes are also presented.


Valetti, F., Morra, S., Barbieri, L., Dezzani, S., Ratto, A., Catucci, G., …Gilardi, G. (2024). Oxygen-resistant [FeFe]hydrogenases: new biocatalysis tools for clean energy and cascade reactions. Faraday Discussions,

Journal Article Type Article
Acceptance Date Feb 21, 2024
Online Publication Date Feb 21, 2024
Publication Date 2024
Deposit Date Apr 19, 2024
Journal Faraday Discussions
Print ISSN 1359-6640
Electronic ISSN 1364-5498
Publisher Royal Society of Chemistry
Peer Reviewed Peer Reviewed
Keywords Physical and Theoretical Chemistry
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