Purification, characterization, and crystallization of monoamine oxidase from escherichia coli k-12

Jung Hyeob Roh, Hideyuki Suzuki, Hiroyuki Azakami, Mitsuo Yamashita, Yoshikatsu Murooka, Hidehiko Kumagai

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42 Citations (Scopus)


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°C. The enzyme was inhibited by a MAO specific inhibitor, hydroxylamine, hydrazine, phenelzine, isoniazid, and tranycypromine. The enzyme oxidized tyramine, phenethylamine, and tryptamine at higher rates, but not oxidized diamine and polyamines such as putrecine and spermine. The antibody against E. coli MAO cross-reacted with purified MAO A from Klebsiella aerogenes.

Original languageEnglish
Pages (from-to)1652-1656
Number of pages5
JournalBioscience, Biotechnology and Biochemistry
Issue number9
Publication statusPublished - 1994 Jan
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry


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