Regulation of the Marinomonas mediterranea antimicrobial protein lysine oxidase by L-lysine and the sensor histidine kinase PpoS.
Some Gram-negative bacteria express a novel enzyme with lysine-epsilon-oxidase (LOD) activity (EC 1.4.3.20). The oxidation of l-Lys generates, among other products, hydrogen peroxide, which confers antimicrobial properties to this kind of enzyme and has been shown to be involved in cell death during biofilm development and differentiation. In addition to LOD, the melanogenic marine bacterium Marinomonas mediterranea, which forms part of the microbiota of the marine plant Posidonia oceanica, expresses two other oxidases of biotechnological interest, a multicopper oxidase, PpoA, with laccase activity and a tyrosinase named PpoB, which is responsible for melanin synthesis. By using both lacZ fusions with the lodAB promoter and quantitative reverse transcription-PCR (qRT-PCR), this study shows that the hybrid sensor histidine kinase PpoS regulates LOD activity at the transcriptional level. Although PpoS also regulates PpoA and PpoB, in this case, the regulatory effect cannot be attributed only to a transcriptional regulation. Further studies indicate that LOD activity is induced at the posttranscriptional level by l-Lys as well as by two structurally similar compounds, l-Arg and meso-2,6-diaminopimelic acid (DAP), neither of which is a substrate of the enzyme. The inducing effect of these compounds is specific for LOD activity since PpoA and PpoB are not affected by them. This study offers, for the first time, insights into the mechanisms regulating the synthesis of the antimicrobial protein lysine-epsilon-oxidase in M. mediterranea, which could be important in the microbial colonization of the seagrass P. oceanica.
Molina-Quintero
Luisa R
LR
Department of Genetics and Microbiology, Faculty of Biology, University of Murcia, Murcia 30100, Spain.
Lucas-Elío
Patricia
P
Sanchez-Amat
Antonio
A
D035961:Alismatales / Q000382:microbiology*; D015153:Blotting, Western; D004156:Catechol Oxidase / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D000071677:Histidine Kinase; D006861:Hydrogen Peroxide / Q000378:metabolism; D008239:Lysine / Q000378:metabolism*; D053544:Marinomonas / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000096:biosynthesis / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D001616:beta-Galactosidase / Q000378:metabolism
D017931:DNA Primers; D006861:Hydrogen Peroxide; D006899:Mixed Function Oxygenases; D004156:Catechol Oxidase; C100031:lysine monooxygenase; D011494:Protein Kinases; D000071677:Histidine Kinase; D001616:beta-Galactosidase; D008239:Lysine