Purification and characterization of monoamine oxidase from Klebsiella aerogenes

Mitsuo Yamashita, Masashi Sakaue, Nobuhide Iwata, Hiroyuki Sugino, Yoshikatsu Murooka

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

The gene for monoamine oxidase (maoA) from Klebsiella aerogenes W70 has been cloned and the enzyme was overproduced in a soluble form. The enzyme was purified approximately 10-fold to homogeneity. The enzyme has a molecular weight of about 79,000, which is identical to the molecular weight deduced from the nucleotide sequence of the gene for monoamine oxidase. The enzyme had maximum activity at pH 6.0 and 50°C when catalyzing the oxidative deamination of tyramine. The enzyme catalyzed the deamination of β-phenylethylamine, dopamine, tryptamine, and octopamine, but not of diamines, polyamines, or amino acids. The enzyme was inhibited by clorgyline, isoniazid, and carbonyl reagents, but not by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide. The enzyme did not exhibit a typical flavoprotein spectrum, but the enzymatic activity increased linearly with increasing amounts of added copper. The purified enzyme was found to contain 10 g of copper per 79,000 g of the protein. The enzymological properties and the amino acid sequence of the enzyme deduced from the nucleotide sequence of the moaA gene are different from those of known tyramine or monoamine oxidases.

Original languageEnglish
Pages (from-to)289-295
Number of pages7
JournalJournal of Fermentation and Bioengineering
Volume76
Issue number4
DOIs
Publication statusPublished - 1993
Externally publishedYes

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Monoamine Oxidase
Purification
Enzymes
Genes
Nucleotides
Amino acids
Copper
Ethyldimethylaminopropyl Carbodiimide
Molecular weight
Clorgyline
Octopamine
Phenethylamines
Oxidoreductases
Amino Acids
Flavoproteins
Tyramine
Diamines
Isoniazid
Polyamines
Dopamine

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Purification and characterization of monoamine oxidase from Klebsiella aerogenes. / Yamashita, Mitsuo; Sakaue, Masashi; Iwata, Nobuhide; Sugino, Hiroyuki; Murooka, Yoshikatsu.

In: Journal of Fermentation and Bioengineering, Vol. 76, No. 4, 1993, p. 289-295.

Research output: Contribution to journalArticle

Yamashita, Mitsuo ; Sakaue, Masashi ; Iwata, Nobuhide ; Sugino, Hiroyuki ; Murooka, Yoshikatsu. / Purification and characterization of monoamine oxidase from Klebsiella aerogenes. In: Journal of Fermentation and Bioengineering. 1993 ; Vol. 76, No. 4. pp. 289-295.
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