The Antimicrobial Activity of Marinocine, Synthesized by Marinomonas mediterranea , Is Due to Hydrogen Peroxide Generated by Its Lysine Oxidase Activity

Marinocine is a broad-spectrum antibacterial protein synthesized by the melanogenic marine bacteriumMarinomonas mediterranea . This work describes the basis for the antibacterial activity of marinocine and the identification of the gene coding for this protein. The antibacterial activity is inhibited under anaerobic conditions and by the presence of catalase under aerobic conditions. Marinocine is active only in culture media containingl -lysine. In the presence of this amino acid, marinocine generates hydrogen peroxide, which causes cell death as confirmed by the increased sensitivity to marinocine ofEscherichia coli strains mutated in catalase activity. The gene coding for this novel enzyme was cloned using degenerate PCR with primers designed based on conserved regions in the antimicrobial protein AlpP, synthesized byPseudoalteromonas tunicata , and some hypothetical proteins. The gene coding for marinocine has been namedlodA , standing for lysine oxidase, and it seems to form part of an operon with a second gene,lodB , that codes for a putative dehydrogenase flavoprotein. The identity of marinocine as LodA has been demonstrated by N-terminal sequencing of purified marinocine and generation oflodA mutants that lose their antimicrobial activity. This is the first report on a bacterial lysine oxidase activity and the first time that a gene encoding this activity has been cloned.