Characterization and a role of Pseudomonas aeruginosa spermidine dehydrogenase in polyamine catabolism

Abstract

Pseudomonas aeruginosaPAO1 has two possible catabolic pathways of spermidine and spermine; one includes thespuAandspuBproducts with unknown functions and the other involves spermidine dehydrogenase (SpdH; EC 1.5.99.6) encoded by an unknown gene. The properties of SpdH inP. aeruginosaPAO1 were characterized and the correspondingspdHgene in this strain identified. The deduced SpdH (620 residues, calculatedMrof 68 861) had a signal sequence of 28 amino acids at the amino terminal and a potential transmembrane segment between residues 76 and 92, in accordance with membrane location of the enzyme. Purified SpdH oxidatively cleaved spermidine into 1,3-diaminopropane and 4-aminobutyraldehyde with a specific activity of 37 units (mg protein)−1and aKmvalue of 36 μM. The enzyme also hydrolysed spermine into spermidine and 3-aminopropanaldehyde with a specific activity of 25 units (mg protein)−1and aKmof 18 μM. Knockout ofspdHhad no apparent effect on the utilization of both polyamines, suggesting that this gene is minimally involved in polyamine catabolism. However, whenspdHwas fused to the polyamine-inducible promoter ofspuA, it fully restored the ability of aspuAmutant to utilize spermidine. It is concluded that SpdH can perform a catabolic rolein vivo, butP. aeruginosaPAO1 does not produce sufficient amounts of the enzyme to execute this function.