Distribution of amine oxidases and amine dehydrogenases in bacteria grown on primary amines and characterization of the amine oxidase from Klebsiella oxytoca

Abstract

The bacteria Klebsiella oxytoca LMD 72.65 (ATCC 8724), Arthrobacter P I LMD 81.60 (NCIB 11 625), Paracoccus wersutus LMD 80.62 (ATCC 25364), Escherichia coli W LMD 50.28 (ATCC 9637), E. coli K12 LMD 93.68, Pseudomonas aeruginosa PA01 LMD 89.1 (ATCC 17933) and Pseudomonas putida LMD 68.20 (ATCC 12633) utilized primary amines as a carbon and energy source, although the range of amines accepted varied from organism t o organism. The Gram-negative bacteria K. oxytoca and E. coli as well as the Gram-positive methylotroph Arthrobacter P I used an oxidase whereas the pseudomonads and the Gramnegative methylotroph Paracoccus wersutus used a dehydrogenase for amine oxidation. K. oxytoca utilized several primary amines but showed a preference for those containing a phenyl group moiety. Only a single oxidase was used for oxidation of the amines. After purification, the following characteristics of the enzyme indicated that it belonged t o the group of copper-quinoprotein amine oxidases (EC 1.4.3.6) : the molecular mass (172 000 Da) of the homodimeric protein; the UWvisible and EPR spectra of isolated and pnitrophenylhydrazine-inhibited enzyme; the presence and the content of copper and topaquinone (TPQ). The amine oxidase appeared to be soluble and localized in the periplasm, but catalase and NAD-dependent aromatic aldehyde dehydrogenase, enzymes catalysing the conversion of its reaction products, were found in the cytoplasm. From the amino acid sequence of the N-terminal part as well as that of a purified peptide, it appears that K. oxytoca produces a copper-quinoprotein oxidase which is very similar to that found in other Enterobacteriaceae.

Production of 2-phenylethylamine by decarboxylation of L-phenylalanine in alkaliphilic Bacillus cohnii.

“…2-Phenylethylamine production has been reported in some gram-negative bacteria belonging to the family Enterobacteriaceae (Ghenghesh and Drucker, 1989). An amine oxidase acting on 2-phenylethylamine occurs in the gram-negatives, Escherichia coli and Klebsiella oxytoca (Hacisalihoglu et al, 1997;Hanlon et al, 1997) and gram-positive Arthrobacter globiformis (Choi et al, 1995), utilizing the amine as a carbon and energy source. The role of the large amount of production of 2-phenylethylamine for the growth of B. cohnii is not clear.…”

Evolution of Catabolic Pathways: Genomic Insights into Microbial s -Triazine Metabolism

“…Interestingly, a significant number of bacteria isolated for growth on alkyl or aromatic amines and polyamines have been Arthrobacter strains (15,24,26,31,38,45,82). Other high-GϩC gram-positive bacteria, Pseudomonas spp., and Klebsiella spp., have also been identified for their metabolism of amines (14,25).…”

Biodegradation of Aromatic Compounds by Escherichia coli

“…The periplasmic MaoA protein (757 aa) shows 84% amino acid sequence identity to the equivalent soluble monoamine oxidase from Klebsiella aerogenes, although the similarity to amino oxidases of gram-positive bacteria and eukaryotic organisms is much lower (124). The MaoA enzyme is a member of the copper-topaquinone (2,4,5-trihydroxyphenylalanine quinone [TPQ]) family of amine oxidases that catalyzes the two-electron oxidative deamination of amines.…”