Characterization of membrane-bound spermidine dehydrogenase of Citrobacter freundii.
Spermidine dehydrogenase found in the membrane fraction of Citrobacter freundii IFO 12681 was solubilized with Triton X-100 and further purified to homogeneity. The properties of the membrane enzyme were almost identical to those obtained from the soluble fraction of the organism with respect to molecular and catalytic properties. Thus, binding properties of the enzyme to the bacterial membrane were checked. The ratio of enzyme activity found in the soluble fraction to the membrane fraction was dependent on salt concentration during cell disruption. A hydrophobic interaction was largely involved in anchoring the enzyme to the membrane fraction. Purified spermidine dehydrogenase from the soluble fraction was readily adsorbed into the membrane fraction in the presence of salt. Spermidine dehydrogenase appeared to be a membrane-bound enzyme localized in the cytoplasmic membranes in a manner that makes a partial release of the enzyme possible during mechanical cell disruption. When spermidine oxidation was done with the resting cells of C. freundii, a stoichiometric formation of two reaction products, 1,3-diaminopropane and gamma-aminobutyraldeyde, was observed without any lag time. These facts indicate that the enzyme is localized on the outer surface of the cytoplasmic membranes or in the periplasmic space of the organism.
Hisano
T
T
Research Institute for Food Science, Kyoto University, Japan.
Murata
K
K
Kimura
A
A
Matsushita
K
K
Toyama
H
H
Adachi
O
O
D000596:Amino Acids / Q000032:analysis; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000201:enzymology; D002848:Chromatography, DEAE-Cellulose / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D016971:Citrobacter freundii / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D017830:Octoxynol; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000302:isolation & purification / Q000378:metabolism*; D011092:Polyethylene Glycols
D000596:Amino Acids; D003902:Detergents; D007527:Isoenzymes; D011092:Polyethylene Glycols; D017830:Octoxynol; D000587:Oxidoreductases Acting on CH-NH Group Donors; C022646:spermidine dehydrogenase