Purification, Characterization, and Crystallization of Monoamine Oxidase from Escherichia coliK-12

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

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

The gene for monoamine oxidase (MAO) was cloned from an Escherichia coligenomic 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. Crystalswere 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. coliMAO cross-reacted with purified MAO A from Klebsiella aerogenes.

Original languageEnglish
Pages (from-to)1652-1656
Number of pages5
JournalBioscience, Biotechnology, and Biochemistry
Volume58
Issue number9
DOIs
Publication statusPublished - 1994
Externally publishedYes

Fingerprint

Monoamine Oxidase
Crystallization
Purification
hydrazine
Enzymes
Phenelzine
DEAE-Cellulose
Column chromatography
Tyramine
Hydroxylamine
Diamines
Spermine
Isoniazid
Polyamines
Ammonium Sulfate
Fractionation
Dimers
Genes
Molecular weight
Antibodies

ASJC Scopus subject areas

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

Cite this

Purification, Characterization, and Crystallization of Monoamine Oxidase from Escherichia coliK-12. / Roh, Jung Hyeob; Suzuki, Hideyuki; Azakami, Hiroyuki; Yamashita, Mitsuo; Murooka, Yoshikatsu; Kumagai, Hidehiko.

In: Bioscience, Biotechnology, and Biochemistry, Vol. 58, No. 9, 1994, p. 1652-1656.

Research output: Contribution to journalArticle

Roh, Jung Hyeob ; Suzuki, Hideyuki ; Azakami, Hiroyuki ; Yamashita, Mitsuo ; Murooka, Yoshikatsu ; Kumagai, Hidehiko. / Purification, Characterization, and Crystallization of Monoamine Oxidase from Escherichia coliK-12. In: Bioscience, Biotechnology, and Biochemistry. 1994 ; Vol. 58, No. 9. pp. 1652-1656.
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