Properties and functions of two succinic-semialdehyde dehydrogenases from Pseudomonas putida.
Two forms of succinic-semialdehyde dehydrogenase have been isolated in Pseudomonas putida. The two enzymes could be separated by filtration on Sephacryl S-300 and their apparent molecular weights were approx. 200,000 and 100,000. The smaller enzyme, which is induced by growth on 4-hydroxyphenylacetate, has been purified to 88% homogeneity by anion-exchange and affinity chromatography. Electrophoresis in sodium dodecyl sulphate gave rise to a molecular weight of 53,000, indicating that the native enzyme is dimeric. Under standard assay conditions this enzyme acts preferentially with NAD but reduces NADP at 9% of the rate observed for NAD. The large enzyme, which is dependent on NADP, is induced by growth on putrescine and its induction is highly coordinated with putrescine: 2-oxoglutarate transaminase, gamma-amino-butyraldehyde dehydrogenase and gamma-aminobutyrate: 2-oxoglutarate transaminase activities. Activity and stability conditions and true Km values for substrate and cosubstrates of the two enzymes were determined.
Sànchez
M
M
Departamento de Bioquìmica, Facultad de Veterinaria, Universidad Complutense, Madrid, Spain.
Alvarez
M A
MA
Balaña
R
R
Garrido-Pertierra
A
A
D002413:Cations, Divalent; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006884:Hydroxybutyrate Dehydrogenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D009243:NAD / Q000494:pharmacology; D009249:NADP / Q000494:pharmacology; D011549:Pseudomonas / Q000201:enzymology*; D011700:Putrescine / Q000378:metabolism; D013696:Temperature; D005680:gamma-Aminobutyric Acid / Q000031:analogs & derivatives / Q000378:metabolism
D002413:Cations, Divalent; D009243:NAD; D009249:NADP; D005680:gamma-Aminobutyric Acid; D006884:Hydroxybutyrate Dehydrogenase; C019416:4-hydroxybutyrate dehydrogenase; C009338:succinic semialdehyde; D011700:Putrescine