4-N-trimethylaminobutyraldehyde dehydrogenase: purification and characterization of an enzyme from Pseudomonas sp. 13CM.
4-N-trimethylaminobutyraldehyde dehydrogenase from Pseudomonas sp. 13CM was purified 14-fold to apparent homogeneity by hydrophobic chromatography on a Phenyl-Toyopearl, and affinity chromatography was done on a 5'-AMP Sepharose4B in the presence of dithiothreitol. The enzyme was found to be a trimer with identical 55 kDa subunits. The isoeletric point was found to be 5.5. The optimum temperature and pH were 40 degrees C and pH 10.0. The purified enzyme was further characterized with respect to substrate specificity, kinetic parameters, and analog inhibition. The K(m) values for 4-N-trimethylaminobutyraldehyde, 4-dimethylaminobutyraldehyde, and NAD(+) were 7.4, 51, and 125 microM respectively. The enzyme was inhibited by SH reagents, and by heavy metal ions.
Hassan
Maizom
M
United Graduate School of Agricultural Sciences, Tottori University, Koyama, Tottori, Japan.
Okada
Masahiro
M
Ichiyanagi
Tsuyoshi
T
Mori
Nobuhiro
N
D000445:Aldehyde Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D007202:Indicators and Reagents; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D011549:Pseudomonas / Q000201:enzymology*; D012685:Sepharose
D001426:Bacterial Proteins; D007202:Indicators and Reagents; D012685:Sepharose; C023095:4-N-trimethylaminobutyraldehyde dehydrogenase; D000445:Aldehyde Oxidoreductases