The antibiotic korormicin A kills bacteria by producing reactive oxygen species

Korormicin is an antibiotic, produced by some Pseudoalteromonads , which selectively kills gram negative bacteria that express the Na + -pumping NADH:quinone oxidoreductase (Na + -NQR.) We show that, although korormicin is an inhibitor of Na + -NQR, the antibiotic action is not a direct result of inhibiting enzyme activity. Instead, perturbation of electron transfer inside the enzyme promotes a reaction between O 2 and one or more redox cofactors in the enzyme (likely the FAD and 2Fe-2S center) leading to the production of reactive oxygen species (ROS). All Pseudoalteromonas contain the nqr operon in their genome including Pseudoalteromonas J010, which produces korormicin. We present activity data indicating that this strain expresses an active Na + -NQR, and that this enzyme is not susceptible to korormicin inhibition. On the basis of our DNA sequence data, we show, that the Na + -NQR of Pseudoalteromonas J010 carries an amino acid substitution (NqrB-G141L, Vibrio cholerae numbering) that, in other Na + -NQR9s confers resistance against korormicin. This is likely the reason that a functional Na + -NQR is able to exist in a bacterium that produces a compound that typically inhibits this enzyme and causes cell death. Korormicin is an effective antibiotic against pathogens such as Vibrio cholerae , Allivibrio fischeri , and Pseudomonas aeruginosa but has no effect on Bacteroides fragilis and thetaiotaomicron, microorganisms that are important members of the human intestinal microflora making it a potentially selective and safe antibiotic. Importance As multi-drug antibiotic resistance in pathogenic bacteria continues to rise, there is a critical need for novel antimicrobial agents. An essential requirement for a useful antibiotic is that it selectively targets bacteria without significant effects on the eukaryotic hosts. Korormicin is an excellent candidate in this respect, because it targets a unique respiratory enzyme found only in prokaryotes, the Na + -pumping NADH:quinone oxidoreductase (Na + -NQR). Korormicin is synthesized by some species of the marine bacterium P seudoalteromonas and is a potent and specific inhibitor of Na + -NQR, an enzyme that is essential for the survival and proliferation of many Gram-negative human pathogens, including Vibrio cholerae , Pseudomonas aeruginosa among others. Here we identified how korormicin selectively kills these bacteria. The binding of korormicin to Na + -NQR promotes the formation of reactive oxygen species generated by the reaction of the FAD and the 2Fe-2S center cofactors with O 2 .