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 bacterium Marinomonas 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 containing l-lysine. In the presence of this amino acid, marinocine generates hydrogen peroxide, which causes cell death as confirmed by the increased sensitivity to marinocine of Escherichia 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 by Pseudoalteromonas tunicata, and some hypothetical proteins. The gene coding for marinocine has been named lodA, 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 of lodA 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.
Department of Genetics and Microbiology, Faculty of Biology, University of Murcia, 30100 Murcia, Spain.
Patricia
P
Lucas-Elío
Daniel
D
Gómez
Francisco
F
Solano
Antonio
A
Sanchez-Amat
D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D003001:Cloning, Molecular; D005798:Genes, Bacterial / Q000235:genetics; D006861:Hydrogen Peroxide / Q000378:metabolism*; D008969:Molecular Sequence Data; D044130:Oceanospirillaceae / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA
D011506:Proteins; C509674:marinocine protein, Marinomonas mediterranea; D006861:Hydrogen Peroxide; D000594:Amino Acid Oxidoreductases; C041123:L-lysine oxidase; D010100:Oxygen