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

Francesca D'Angelo, Elena Fernández-Fueyo, Pierre Simon Garcia, Helena Shomar, R.S. Cebola Rebelo Manuel, Ferhat Büke, Niels van den Broek, Carol de Ram, Martin Pabst, More Authors

Research output: Contribution to journalArticleScientificpeer-review

6 Citations (Scopus)
26 Downloads (Pure)


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.

Original languageEnglish
Article numbere70936
Number of pages26
Publication statusPublished - 2022


  • biochemistry
  • chemical biology
  • electron transfer protein
  • escherichia coli
  • horizontal gene transfer
  • infectious disease
  • iron-sulfur enzyme
  • microbial engineering
  • microbiology


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