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Cellular assays identify barriers impeding iron-sulfur enzyme activity in a non-native prokaryotic host

Abstract : Iron-sulfur (Fe-S) clusters are ancient and ubiquitous protein cofactors and play irreplaceable roles in many metabolic and regulatory processes. Fe-S clusters are built and distributed to Fe-S enzymes by dedicated protein networks. The core components of these networks are widely conserved and highly versatile. However, Fe-S proteins and enzymes are often inactive outside their native host species. We sought to systematically investigate the compatibility of Fe-S networks with non-native Fe-S enzymes. By using collections of Fe-S enzyme orthologs representative of the entire range of prokaryotic diversity, we uncovered a striking correlation between phylogenetic distance and probability of functional expression. Moreover, coexpression of a heterologous Fe-S biogenesis pathway increases the phylogenetic range of orthologs that can be supported by the foreign host. We also find that Fe-S enzymes that require specific electron carrier proteins are rarely functionally expressed unless their taxon-specific reducing partners are identified and co-expressed. We demonstrate how these principles can be applied to improve the activity of a radical S-adenosyl methionine(rSAM) enzyme from a Streptomyces antibiotic biosynthesis pathway in Escherichia coli. Our results clarify how oxygen sensitivity and incompatibilities with foreign Fe-S and electron transfer networks each impede heterologous activity. In particular, identifying compatible electron transfer proteins and heterologous Fe-S biogenesis pathways may prove essential for engineering functional Fe-S enzyme-dependent pathways.
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Submitted on : Monday, April 11, 2022 - 11:19:45 AM
Last modification on : Friday, August 5, 2022 - 12:03:01 PM
Long-term archiving on: : Tuesday, July 12, 2022 - 6:20:22 PM


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Francesca D’angelo, Elena Fernández-Fueyo, Pierre Simon Garcia, Héléna Shomar, Martin Pelosse, et al.. Cellular assays identify barriers impeding iron-sulfur enzyme activity in a non-native prokaryotic host. eLife, eLife Sciences Publication, 2022, 11, pp.e70936. ⟨10.7554/eLife.70936⟩. ⟨hal-03634503⟩



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