Multiple and interconnected pathways for L-lysine catabolism in Pseudomonas putida KT2440.
L-lysine catabolism in Pseudomonas putida KT2440 was generally thought to occur via the aminovalerate pathway. In this study we demonstrate the operation of the alternative aminoadipate pathway with the intermediates D-lysine, L-pipecolate, and aminoadipate. The simultaneous operation of both pathways for the use of L-lysine as the sole carbon and nitrogen source was confirmed genetically. Mutants with mutations in either pathway failed to use L-lysine as the sole carbon and nitrogen source, although they still used L-lysine as the nitrogen source, albeit at reduced growth rates. New genes were identified in both pathways, including the davB and davA genes that encode the enzymes involved in the oxidation of L-lysine to delta-aminovaleramide and the hydrolysis of the latter to delta-aminovalerate, respectively. The amaA, dkpA, and amaB genes, in contrast, encode proteins involved in the transformation of Delta1-piperidine-2-carboxylate into aminoadipate. Based on L-[U-13C, U-15N]lysine experiments, we quantified the relative use of pathways in the wild type and its isogenic mutants. The fate of 13C label of L-lysine indicates that in addition to the existing connection between the D- and L-lysine pathways at the early steps of the catabolism of L-lysine mediated by a lysine racemase, there is yet another interconnection at the lower end of the pathways in which aminoadipate is channeled to yield glutarate. This study establishes an unequivocal relationship between gene and pathway enzymes in the metabolism of L-lysine, which is of crucial importance for the successful colonization of the rhizosphere of plants by this microorganism.
Revelles
Olga
O
Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, C/Prof. Albareda 1, 18008 Granada, Spain.
Espinosa-Urgel
Manuel
M
Fuhrer
Tobias
T
Sauer
Uwe
U
Ramos
Juan L
JL
D015074:2-Aminoadipic Acid / Q000378:metabolism; D000591:Amino Acid Isomerases; D001426:Bacterial Proteins / Q000235:genetics; D002244:Carbon / Q000378:metabolism; D002250:Carbon Radioisotopes / Q000378:metabolism; D004251:DNA Transposable Elements; D004798:Enzymes / Q000235:genetics; D008401:Gas Chromatography-Mass Spectrometry; D005798:Genes, Bacterial; D005977:Glutarates / Q000378:metabolism; D008239:Lysine / Q000378:metabolism*; D016254:Mutagenesis, Insertional; D009154:Mutation; D009584:Nitrogen / Q000378:metabolism; D010875:Pipecolic Acids / Q000378:metabolism; D010880:Piperidines; D016958:Pseudomonas putida / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D014631:Valerates / Q000378:metabolism
D001426:Bacterial Proteins; D002250:Carbon Radioisotopes; D004251:DNA Transposable Elements; D004798:Enzymes; D005977:Glutarates; D010875:Pipecolic Acids; D010880:Piperidines; D014631:Valerates; D015074:2-Aminoadipic Acid; D002244:Carbon; D000591:Amino Acid Isomerases; C084211:lysine racemase; C031345:pipecolic acid; D008239:Lysine; D009584:Nitrogen