Functional expression of amine oxidase from Aspergillus niger (AO-I) in Saccharomyces cerevisiae.
The aim of this work was to prepare recombinant amine oxidase from Aspergillus niger after overexpressing in yeast. The yeast expression vector pDR197 that includes a constitutive PMA1 promoter was used for the expression in Saccharomyces cerevisiae. Recombinant amine oxidase was extracted from the growth medium of the yeast, purified to homogeneity and identified by activity assay and MALDI-TOF peptide mass fingerprinting. Similarity search in the newly published A. niger genome identified six genes coding for copper amine oxidase, two of them corresponding to the previously described enzymes AO-I a methylamine oxidase and three other genes coding for FAD amine oxidases. Thus, A. niger possesses an enormous metabolic gear to grow on amine compounds and thus support its saprophytic lifestyle.
Kolaríková
Katerina
K
Department of Biochemistry, Palacký University, Slechtitelů 11, 783 71, Olomouc, Czech Republic.
Galuszka
Petr
P
Sedlárová
Iva
I
Sebela
Marek
M
Frébort
Ivo
I
D006631:Amine Oxidase (Copper-Containing) / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000588:Amines / Q000378:metabolism; D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D002848:Chromatography, DEAE-Cellulose; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment
D000588:Amines; D005656:Fungal Proteins; D011994:Recombinant Proteins; C007940:hexylamine; D006631:Amine Oxidase (Copper-Containing)