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

eLife

Helena Shomar¹  Sandrine Ollagnier de Choudens²  Niels van den Broek²  Ferhat Büke³  Elena Fernández-Fueyo³  Martin Pelosse³  Nicolas Duraffourg³  Siyi Liu³  Emmanuelle Bouveret⁴  Martin Pabst⁴  Béatrice Py⁵  Francesca D'Angelo⁶  Gregory Bokinsky⁶  Pierre Simon Garcia⁶  Simonetta Gribaldo⁶  Rita Rebelo Manuel⁷  Frédéric Barras⁷  Carol de Ram⁷ 

[1] Institut Pasteur, Université de Paris, CNRS UMR6047, Evolutionary Biology of the Microbial Cell, Department of Microbiology, Paris, France;Aix-Marseille Université-CNRS, Laboratoire de Chimie Bactérienne UMR 7283, Institut de Microbiologie de la Méditerranée, Institut Microbiologie Bioénergies Biotechnologie, Marseille, France;Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Delft, Netherlands;Department of Biotechnology, Delft University of Technology, Delft, Netherlands;Institut Pasteur, Université de Paris, CNRS UMR6047, Evolutionary Biology of the Microbial Cell, Department of Microbiology, Paris, France;Unit Stress Adaptation and Metabolism of Enterobacteria, Department of Microbiology, Université de Paris, UMR CNRS 2001, Institut Pasteur, Paris, France;Univ. Grenoble Alpes, CNRS, CEA, IRIG, Laboratoire de Chimie et Biologie des Métaux, Grenoble, France;

关键词: iron-sulfur enzyme;    horizontal gene transfer;    microbial engineering;    electron transfer protein;   

DOI  :  10.7554/eLife.70936

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown