Oxydifficidin, a potentNeisseria gonorrhoeaeantibiotic due to DedA assisted uptake and ribosomal protein RplL sensitivity

Gonorrhea, which is caused byNeisseria gonorrhoeae, is the second most reported sexually transmitted infection worldwide. The increasing appearance of isolates that are resistant to approved therapeutics raises the concern that gonorrhea may become untreatable. Here, we serendipitously identified oxydifficidin as a potentN. gonorrhoeaeantibiotic through the observation of aBacillus amyloliquefacienscontaminant in a lawn ofN. gonorrhoeae. Oxydifficidin is active against both wild- type and multidrug-resistantN. gonorrhoeae. It’s potent activity results from a combination of DedA- assisted uptake into the cytoplasm and the presence of an oxydifficidin-sensitive ribosomal protein L7/L12 (RplL). Our data indicates that oxydifficidin binds to the ribosome at a site that is distinct from other antibiotics and that L7/L12 is uniquely associated with its mode of action. This study opens a potential new avenue for addressing antibiotic resistant gonorrhea and underscores the possibility of identifying overlooked natural products from cultured bacteria, particularly those with activity against previously understudied pathogens.