title: Purification, characterization, and crystallization of monoamine oxidase from Escherichia coli K-12.
abstract: The gene for monoamine oxidase (MAO) was cloned from an Escherichia coli genomic library and MAO was overproduced in the periplasmic space. The enzyme was purified to homogeneity by preparation of a periplasmic fraction, followed by ammonium sulfate fractionation and DEAE-cellulose column chromatography. Crystals were obtained by the hanging drop method using sodium citrate as a precipitant. The enzyme was found to be a dimer of identical subunits with a molecular weight of 80,000, and showed the highest activity at pH 7.5 and 45 degrees C. The enzyme was inhibited by a MAO specific inhibitor, hydroxylamine, hydrazine, phenelzine, isoniazid, and tranycpromine. The enzyme oxidized tyramine, phenethylamine, and tryptamine at higher rates, but not oxidized diamine and polyamines such as putrescine and spermine. The antibody against E. coli MAO cross-reacted with purified MAO A from Klebsiella aerogenes.
authors.affiliation: Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Japan.
authors.forename: J H
authors.initials: JH
authors.lastname: Roh
authors.forename: H
authors.initials: H
authors.lastname: Suzuki
authors.forename: H
authors.initials: H
authors.lastname: Azakami
authors.forename: M
authors.initials: M
authors.lastname: Yamashita
authors.forename: Y
authors.initials: Y
authors.lastname: Murooka
authors.forename: H
authors.initials: H
authors.lastname: Kumagai
mesh_terms: D006631:Amine Oxidase (Copper-Containing) / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D003001:Cloning, Molecular; D003460:Crystallization; D004926:Escherichia coli / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008995:Monoamine Oxidase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D008996:Monoamine Oxidase Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D008996:Monoamine Oxidase Inhibitors; D006631:Amine Oxidase (Copper-Containing); D008995:Monoamine Oxidase