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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 δ-aminovaleramide and the hydrolysis of the latter to δ-aminovalerate, respectively. The amaA, dkpA, and amaB genes, in contrast, encode proteins involved in the transformation of Δ¹ …