Induction and characterization of a novel amine oxidase from the yeast Kluyveromyces marxianus.
An amine oxidase from the yeast Kluyveromyces marxianus was induced, purified and completely characterized; it was shown to belong to the class of copper-containing amine oxidases (E.C. 1.4.3.6). The enzyme was induced by putrescine and, very strongly, by copper(II); structural-functional characterization of the enzyme was performed, including determination of molecular weight, glycosylation, copper and TPQ content, isoelectric point, K(M) and k(CAT) (with benzylamine as substrate), pH, temperature and ionic strength effect on catalysis, substrate and inhibitor specificity. A 700 bp clone was isolated containing the cDNA that encodes for the C-terminus of the enzyme; the amino acid sequence deduced (the first available for a benzylamine oxidase from yeast) was compared to that of other copper amine oxidases from microorganisms and higher organisms. From the results obtained, the putrescine/benzylamine oxidase from Kluyveromyces marxianus was found to have a good homology with other enzymes of this class from microorganisms, and particularly with AO I from Aspergillus niger. Nonetheless, some features resulted closer to those of animal amine oxidases and histaminases. Some potential biotechnological applications are proposed. The cDNA Accession No. is AJ320485.
Corpillo
Davide
D
LIMA, BioIndustry Park del Canavese, Colleretto Giacosa, Torino, Italy.
Valetti
Francesca
F
Giuffrida
Maria Gabriella
MG
Conti
Amedeo
A
Rossi
Antonello
A
Finazzi-Agrò
Alessandro
A
Giunta
Carlo
C
D006631:Amine Oxidase (Copper-Containing) / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D001595:Benzylamine Oxidase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002848:Chromatography, DEAE-Cellulose; D003300:Copper / Q000494:pharmacology; D004295:Dihydroxyphenylalanine / Q000031:analogs & derivatives* / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction / Q000187:drug effects; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007700:Kinetics; D007716:Kluyveromyces / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011700:Putrescine / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D014462:Ultrafiltration
D004295:Dihydroxyphenylalanine; C067626:6-hydroxydopa quinone; D003300:Copper; D001595:Benzylamine Oxidase; D006631:Amine Oxidase (Copper-Containing); D011700:Putrescine