Isolation and characterization of a new [FeFe]-hydrogenase from Clostridium perfringens

Morra, Simone; Mongili, Beatrice; Maurelli, Sara; Gilardi, Gianfranco; Valetti, Francesca

Abstract

© 2015 International Union of Biochemistry and Molecular Biology, Inc. This paper reports the first characterization of an [FeFe]-hydrogenase from a Clostridium perfringens strain previously isolated in our laboratory from a pilot-scale bio-hydrogen plant that efficiently produces H2 from waste biomasses. On the basis of sequence analysis, the enzyme is a monomer formed by four domains hosting various iron–sulfur centres involved in electron transfer and the catalytic center H-cluster. After recombinant expression in Escherichia coli, the purified protein catalyzes H2 evolution at high rate of 1645±16s−1. The optimal conditions for catalysis are in the pH range 6.5–8.0 and at the temperature of 50°C. EPR spectroscopy showed that the H-cluster of the oxidized enzyme displays a spectrum coherent with the Hox state, whereas the CO-inhibited enzyme has a spectrum coherent with the Hox-CO state. FTIR spectroscopy showed that the purified enzyme is composed of a mixture of redox states, with a prevalence of the Hox; upon reduction with H2, vibrational modes assigned to the Hred state were more abundant, whereas binding of exogenous CO resulted in a spectrum assigned to the Hox-CO state. The spectroscopic features observed are similar to those of the [FeFe]-hydrogenases class, but relevant differences were observed given the different protein environment hosting the H-cluster.

Citation

Morra, S., Mongili, B., Maurelli, S., Gilardi, G., & Valetti, F. (2016). Isolation and characterization of a new [FeFe]-hydrogenase from Clostridium perfringens. Biotechnology and Applied Biochemistry, 63(3), 305-311. https://doi.org/10.1002/bab.1382

• Journal Article Type: Article
• Acceptance Date: Apr 7, 2015
• Online Publication Date: Jul 1, 2015
• Publication Date: Jun 17, 2016
• Deposit Date: Jan 31, 2020
• Publicly Available Date: Feb 11, 2020
• Journal: Biotechnology and Applied Biochemistry
• Print ISSN: 0885-4513
• Electronic ISSN: 1470-8744
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 63
• Issue: 3
• Pages: 305-311
• DOI: https://doi.org/10.1002/bab.1382
• Public URL: https://nottingham-repository.worktribe.com/output/3841268
• Publisher URL: https://iubmb.onlinelibrary.wiley.com/doi/abs/10.1002/bab.1382
• Additional Information: This is the peer reviewed version of the following article: Morra, S., Mongili, B., Maurelli, S., Gilardi, G. and Valetti, F. (2016), Isolation and characterization of a new [FeFe]‐hydrogenase from Clostridium perfringens. Biotechnology and Applied Biochemistry, 63: 305-311, which has been published in final form at https://doi.org/10.1002/bab.1382. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.