Atypical effect of temperature tuning on the insertion of the catalytic iron−sulfur center in a recombinant [FeFe]-hydrogenase

Morra, Simone; Cordara, Alessandro; Gilardi, Gianfranco; Valetti, Francesca

Abstract

© 2015 The Protein Society. The expression of recombinant [FeFe]-hydrogenases is an important step for the production of large amount of these enzymes for their exploitation in biotechnology and for the characterization of the protein-metal cofactor interactions. The correct assembly of the organometallic catalytic site, named H-cluster, requires a dedicated set of maturases that must be coexpressed in the microbial hosts or used for in vitro assembly of the active enzymes. In this work, the effect of the post-induction temperature on the recombinant expression of CaHydA [FeFe]-hydrogenase in E. coli is investigated. The results show a peculiar behavior: the enzyme expression is maximum at lower temperatures (20C), while the specific activity of the purified CaHydA is higher at higher temperature (30C), as a consequence of improved protein folding and active site incorporation.

Citation

Morra, S., Cordara, A., Gilardi, G., & Valetti, F. (2015). Atypical effect of temperature tuning on the insertion of the catalytic iron−sulfur center in a recombinant [FeFe]-hydrogenase. Protein Science, 24(12), 2090-2094. https://doi.org/10.1002/pro.2805

• Journal Article Type: Article
• Acceptance Date: Sep 11, 2015
• Online Publication Date: Sep 24, 2015
• Publication Date: Nov 25, 2015
• Deposit Date: Jan 31, 2020
• Publicly Available Date: Feb 11, 2020
• Journal: Protein Science
• Print ISSN: 0961-8368
• Electronic ISSN: 1469-896X
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 24
• Issue: 12
• Pages: 2090-2094
• DOI: https://doi.org/10.1002/pro.2805
• Public URL: https://nottingham-repository.worktribe.com/output/3841417
• Publisher URL: https://onlinelibrary.wiley.com/doi/full/10.1002/pro.2805
• Additional Information: This is the peer reviewed version of the following article: Morra, S., Cordara, A., Gilardi, G. and Valetti, F. (2015), Atypical effect of temperature tuning on the insertion of the catalytic iron−sulfur center in a recombinant [FeFe]‐hydrogenase. Protein Science, 24: 2090-2094., which has been published in final form at https://doi.org/10.1002/pro.2805. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.