title: Purification and properties of two succinic semialdehyde dehydrogenases from Klebsiella pneumoniae.
abstract: Two forms of succinic semialdehyde dehydrogenase have been isolated in Klebsiella pneumoniae M5a1. The two enzymes could be separated by filtration on Sephacryl S-300 and their apparent molecular weights were approx. 275,000 and 300,000. The large enzyme is specific for NADP. The smaller enzyme, which is induced by growth on 3-hydroxyphenylacetic acid, 4-hydroxyphenylacetic acid, 3,4-dihydroxyphenylacetic acid and gamma-aminobutyrate, has been purified to 96% homogeneity by affinity chromatography. The NAD-linked succinic semialdehyde dehydrogenase was able to use NADP as cofactor. Its induction is coordinated with 3- and 4-hydroxylase, the enzymes which initiate degradation of 3- and 4-hydroxyphenylacetic acid. The NAD-linked form is also induced by exogenous succinic semialdehyde. The large enzyme is specific for NADP and has been isolated from a defective mutant which lacked the activity of the NAD-linked succinic semialdehyde dehydrogenase. Activity and stability conditions and true K m values for substrates and cosubstrates of the two enzymes were determined. Some aspects of the induction of the NAD-linked enzyme participating in the metabolism of 4-hydroxyphenylacetic and gamma-aminobutyrate were studied.
authors.affiliation: Departamento de Bioquímica, Facultad de Veterinaria, Universidad Complutense, Madrid, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Sanchez
authors.forename: J
authors.initials: J
authors.lastname: Fernández
authors.forename: M
authors.initials: M
authors.lastname: Martin
authors.forename: A
authors.initials: A
authors.lastname: Gibello
authors.forename: A
authors.initials: A
authors.lastname: Garrido-Pertierra
mesh_terms: D004790:Enzyme Induction; D006884:Hydroxybutyrate Dehydrogenase / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D007711:Klebsiella pneumoniae / Q000201:enzymology*; D009243:NAD; D009249:NADP
chemical_list: D009243:NAD; D009249:NADP; D006884:Hydroxybutyrate Dehydrogenase; C019416:4-hydroxybutyrate dehydrogenase