Characterization and application of aminoamide-oxidizing enzyme from Aspergillus carbonarius AIU 205
Section snippets
Chemicals
l-Lysine, l-ornithine, aliphatic amines and diamines were purchased from Wako Pure Chemicals (Osaka, Japan). 4-ABAD and plasma amine oxidase were from Toronto Research Chemicals (Toronto, Canada) and Worthington Biochemical Co. (Lakewood, NJ, USA), respectively. Peroxidase was gift from Amano Enzyme (Nagoya, Japan). All other chemicals used were of analytical grade and commercially available.
Screening of microorganism
Fungal strains were incubated in a 500-ml shaker flask containing 100 ml of a n-butylamine medium, pH
Screening of microorganisms
We isolated 12 fungal strains as producers of an enzyme exhibiting oxidase activity for 4-ABAD. The crude enzyme solution from each isolated strain also exhibited oxidase activity for n-butylamine (Table 1). Since it was known that amine oxidase from Aspergillus niger AKU 3302 catalyzed oxidative deamination of aliphatic monoamines, aromatic monoamines and aliphatic diamines 4, 5, we purified the amine oxidase, and assayed the 4-ABAD oxidase activity. The amine oxidase also oxidized 4-ABAD, but
Discussion
Here, we demonstrated a new result that the copper-containing amine oxidase from A. niger AKU 3302 oxidized 4-ABAD as well as amines. However, the amine oxidase was not applicable for l-lysine assay coupling with l-AAO/MOG from Pseudomonas sp. AIU 813, because the Km value for 4-ABAD was high. Since, enzymes with low Km values are generally suitable for coupling enzymes, we selected A. carbonarius AIU 205 as a new producer of enzymes exhibiting oxidase activity for 4-ABAD, and revealed certain
Acknowledgment
This work was financially supported by the Hokuriku Innovation Cluster for Health Science, Ministry of Education, Culture, Sports, Science, and Technology.
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