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

作者：Morra Simone; Cordara Alessandro; Gilardi Gianfranco; Valetti Francesca^*

来源：Protein Science, 2015, 24(12): 2090-2094.

DOI：10.1002/pro.2805

• [FeFe]-hydrogenases
• recombinant expression
• bio-hydrogen
• metalloenzyme

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 (20 degrees C), while the specific activity of the purified CaHydA is higher at higher temperature (30 degrees C), as a consequence of improved protein folding and active site incorporation.

• 出版日期2015-12