Metabolism of Pipecolic Acid in a Pseudomonas Species IV. Electron Transport Particle of Pseudomonas putida

Pseudomonas putida metabolizes D-lysine to Al-piperideine-2-carboxylate and L-pipecolate. The second step of this catabolic pathway is catalyzed by A'piperideine-2-carboxylate reductase. This enzyme was isolated and purified from cells grown on DL-lysine as substrate. The enzyme was very unstable, resulting in low recovery of activity and low purity after a six-step purification procedure. The enzyme had a pH optimum of 8.0 to 8.3. The Km values for A-piperideine-2carboxylate and NADPH were 0.23 and 0.13 mM, respectively. NADPH at concentrations above 0.15 mM was inhibitory to the enzyme. A1-pyrroline-5carboxylate, pyroglutamate, and NADH were poor substrates or coenzyme for Al-piperideine-2-carboxylate reductase. The enzyme reaction from A-piperideine-2-carboxylate to L-pipecolate was irreversible. EDTA, sodium pyrophosphate, and dithiothreitol at concentrations of 1 mM protected the enzyme during storage. The enzyme was inhibited almost totally by Zn2+, Mn2i, Hg2+ Co2+,

Section: Methodsmentioning

confidence: 99%

“…Recently, it has also been shown to be a major route for D-lysine and L-lysine metabolism in rat brains (8)(9)(10)19). Pipecolic acid metabolism in Pseudomonas involving the reaction steps L-pipecolate -* Al-piperideine-6-carboxylate --L-ot-aminoadipate has been studied most definitively by Rodwell and associates (4)(5)(6)(7)36).…”

mentioning

confidence: 99%

delta1-piperideine-2-carboxylate reductase of Pseudomonas putida

Payton¹,

Chang²

1982

“…Bacteriological. Cultures of P. putida P2, grown as described previously (2), with L-lysine (occasionally DL-pipecolate) as sole organic nutrient, were harvested at 60% maximal growth by centrifugation at 10,000 X g and were washed once with 0.05 M phosphate buffer (pH 6.7), and the cells were frozen.…”

Section: Methodsmentioning

confidence: 99%

Metabolism of Pipecolic Acid in a Pseudomonas Species V. Pipecolate Oxidase and Dehydrogenase

Baginsky

Rodwell

1967

Self Cite

Oxidation of pipecolate to Al-piperideine-6-carboxylate is catalyzed by pipecolate oxidase, an inducible, membrane-bound dehydrogenase associated with the electron transport components of Pseudomonas putida P2. From the oxidase, we obtained a smaller particle containing flavine adenine dinucleotide (FAD) and cytochrome b, but no longer able to catalyze electron transfer to oxygen or to cytochrome c. Certain properties of this L-pipecolate dehydrogenase, an FADflavoprotein, are reported.

“…The latter compound is converted to glutarate by transamination and oxidation (10,34). n-Lysine is metabolized to pipecolate (7,22), which is oxidized to 2-aminoadipate and then to gIutaryl-coenzyme A (2,36), the fate of which is known (23). The data to be presented here indicate that the strain ofPseudomonas fluorescens used also oxidizes lysine by these pathways and 5-hydroxylysine by two similar, but different, pathways.…”

mentioning

confidence: 99%

Metabolism of 5-hydroxylysine in Pseudomonas fluorescens

Friede¹,

Henderson²

1976

Hydroxylysine is metabolized via two routes by a Pseudomonas fluorescens strain as shown by the oxidation of selected intermediates. Hydroxy-ilysine is oxidized via a pathway analogous to the monooxygenase pathway for ilysine, and data suggest that at least some of the enzymes are those involved in the metabolism of ilysine. Hydroxy-Irlysine is also converted by a racemase to allohydroxy-D-lysine, which is then degraded via a pathway analogous to, but different from, that described for D-lysine, involving hydroxy-L-pipecolate, 2amino-5-hydroxyadipate, and 2-hydroxyglutarate. Data obtained with mutants unable to oxidize L-pipecolate suggest that the enzymes for the metabolism of hydroxy-L-pipecolate are distinct from those for L-pipecolate. Studies on nand Llysine degradation have shown that the previously described pathways for these compounds are present in this soil pseudomonad.