title: Purification and characterization of 4-N-trimethylamino-1-butanol dehydrogenase of Pseudomonas sp. 13CM.
abstract: A new enzyme, NAD+-dependent 4-N-trimethylamino-1-butanol dehydrogenase from Pseudomonas sp. 13CM, was purified 526-fold to apparent homogeneity in 5 chromatographic steps. The enzyme had a molecular mass of 45 kDa and appeared to be a monomer enzyme. The isoeletric point was found to be 4.8. The optimum temperature was 50 degrees C, and the optimum pHs for the oxidation and reduction reactions were 9.5 and 6.0 respectively. The purified enzyme was further characterized with respect to substrate specificity, kinetic parameters, and amino acid terminal sequence. The Km values for trimethylamino-1-butanol and NAD+ were 0.54 mM and 0.22 mM respectively. In the reduction reaction, the apparent Km values for trimethylaminobutylaldehyde and NADH were 0.67 mM and 0.04 mM, respectively. The enzyme was inhibited by SH reagents, chelating reagents, and heavy metal ions. The N-terminal 12 amino acid residues were sequenced.
authors.affiliation: United Graduate School of Agricultural Sciences, Tottori University, Japan.
authors.forename: Maizom
authors.initials: M
authors.lastname: Hassan
authors.forename: Sachiko
authors.initials: S
authors.lastname: Morimoto
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Murakami
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Ichiyanagi
authors.forename: Nobuhiro
authors.initials: N
authors.lastname: Mori
mesh_terms: D000429:Alcohol Oxidoreductases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D000440:Butanols / Q000378:metabolism; D007700:Kinetics; D010084:Oxidation-Reduction; D011549:Pseudomonas / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D000447:Aldehydes; D001426:Bacterial Proteins; D000440:Butanols; C029041:gamma-trimethylaminobutyraldehyde; D000429:Alcohol Oxidoreductases; C522930:4-N-trimethylamino-1-butanol dehydrogenase, Pseudomonas; C060833:butanol dehydrogenase