Purification and characterization of an L-amino acid oxidase from Pseudomonas sp. AIU 813.
An L-amino acid oxidase was found from a newly isolated strain, Pseudomonas sp. AIU 813. This enzyme was remarkably induced by incubation with L-lysine as a nitrogen source, and efficiently purified using an affinity chromatography with L-lysine as ligand. The enzyme oxidized L-lysine, L-ornithine and L-arginine, but not other L-amino acids and d-amino acids. The oxidase activity for L-lysine was detected in a wide pH range, and its optimal was pH 7.0. In contrast, the oxidase activity for L-ornithine and L-arginine was not shown in acidic region from pH 6.5, and optimal pH for both substrates was 9.0. The enzyme was a flavoprotein and composed of two identical subunits with molecular mass of 54.5 kDa. The N-terminal amino acid sequence was similar to that of putative flavin-containing amine oxidase and putative tryptophan 2-monooxygenase, but not to that of L-amino acid oxidases.
Department of Biological Chemistry and Food Science, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka 020-8550, Japan. kiso@iwate-u.ac.jp
Kimiyasu
K
Isobe
Asami
A
Sugawara
Hanako
H
Domon
Yasuhisa
Y
Fukuta
Yasuhisa
Y
Asano
D000595:Amino Acid Sequence; D001120:Arginine / Q000378:metabolism / Q000494:pharmacology; D003067:Coenzymes / Q000378:metabolism / Q000494:pharmacology; D004790:Enzyme Induction / Q000187:drug effects; D005420:Flavoproteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D050830:L-Amino Acid Oxidase / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009584:Nitrogen / Q000378:metabolism / Q000494:pharmacology; D009952:Ornithine / Q000378:metabolism / Q000494:pharmacology; D011549:Pseudomonas / Q000145:classification / Q000201:enzymology* / Q000254:growth & development / Q000302:isolation & purification; D013379:Substrate Specificity; D013696:Temperature; D013997:Time Factors
D003067:Coenzymes; D005420:Flavoproteins; D001120:Arginine; D009952:Ornithine; D050830:L-Amino Acid Oxidase; D008239:Lysine; D009584:Nitrogen