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

Pipecolic acid oxidase from Rhodotorula glutinis, which converts pipecolic acid to a-aminoadipic-&-semialdehyde, an intermediate of the biosynthetic pathway of lysine, was purified 290-fold. The enzyme from the crude extract and purified preparation exhibited a molecular weight of approximately 43,000 and was composed of a single subunit. The purified enzyme was heat labile and exhibited a pH optimum of 8.5 and an apparent Km for L-pipecolic acid of 1.67 x 10-3 M. L-Proline acted as a competitive inhibitor for the enzyme. The enzyme was inhibited by the sulfhydryl agents p-chloromercuribenzoate and mercuric chloride. The in vitro enzyme activity required oxygen, and upon oxidation of pipecolic acid, oxygen was reduced to hydrogen peroxide.

Section: Discussionmentioning

confidence: 99%

“…For this catabolism, pipecolic acid is converted to ASA by pipecolic acid oxidase. This enzyme is associated with an electron transport particle and has been purified sixfold (2). Since lysine is synthesized in Pseudomonas spp.…”

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confidence: 99%

Lysine biosynthesis in Rhodotorula glutinis: properties of pipecolic acid oxidase

Kinzel¹,

Bhattacharjee²

1982

“…1). The first two enzymes for the metabolism of pipecolate (3,6) catalyze the loss of four electrons in two stages of oxidation, or 0.5 mol of 02 each. Therefore, the mutation in this strain appears to be in 2-aminoadipate transaminase or the subsequent enzyme (Fig.…”

Section: Downloaded Frommentioning

confidence: 99%

“…It was identified as a strain of P. fluorescens as described by Stanier et al (33). The culture was grown at 30°C with shaking on a basal salts medium (2) to which were added 3 mM lysine, 3 mM hydroxylysine, or 10 mM succinate as carbon source and 1.5 mM ammonium chloride as nitrogen source.…”

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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.

“…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).…”

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confidence: 99%

delta1-piperideine-2-carboxylate reductase of Pseudomonas putida

Payton¹,

Chang²

1982

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+,