Properties of the flavoenzyme D-aspartate oxidase from Octopus vulgaris.
The properties of D-aspartate oxidase from Octopus vulgaris (EC 1.4.3.1) have been investigated. The protein is a monomer of M(r) 37,000 containing one mol flavin/mol protein. The enzyme as isolated exists at least in two forms, one containing FAD and the other, which is catalytically inactive, probably containing 6-OH-FAD, as inferred from the absorption spectrum of the enzyme. An additional form of the enzyme, as far as the nature of the coenzyme is concerned, has been detected in the purified enzyme and shown to derive from the form originally containing FAD. The modulation of the coenzyme reactivity exerted by Octopus D-aspartate oxidase, as studied by spectrophotometric techniques, conforms to the one expected for an enzyme belonging to the oxidase class of flavoproteins. Structural investigations show similarities in both the amino-acid composition and the N-terminal amino-acid sequence to bovine D-aspartate oxidase and porcine D-amino-acid oxidase. In summary, the general properties of the enzyme from Octopus vulgaris closely resemble those of the enzyme from beef kidney. Moreover, kinetic analyses suggest that two active-site residues with pKa of 7.1 and 9.1 are critical for catalysis, and that the ionization of such residues has different effects on the catalytic activity depending whether mono- or dicarboxylic D-amino acids are used as substrate.
Tedeschi
G
G
Istituto di Fisiologia Veterinaria e Biochimica, Università di Milano, Italy.
Negri
A
A
Ceciliani
F
F
Ronchi
S
S
Vetere
A
A
D'Aniello
G
G
D'Aniello
A
A
D000594:Amino Acid Oxidoreductases / Q000737:chemistry* / Q000378:metabolism; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D002417:Cattle; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D050804:D-Aspartate Oxidase; D005182:Flavin-Adenine Dinucleotide / Q000031:analogs & derivatives / Q000032:analysis; D005971:Glutamates / Q000378:metabolism; D018698:Glutamic Acid; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D008969:Molecular Sequence Data; D049831:Octopodiformes / Q000201:enzymology*; D013053:Spectrophotometry; D013379:Substrate Specificity; D013552:Swine
D000596:Amino Acids; D005971:Glutamates; D005182:Flavin-Adenine Dinucleotide; D001224:Aspartic Acid; D018698:Glutamic Acid; C052277:6-hydroxy-FAD; D000594:Amino Acid Oxidoreductases; D050804:D-Aspartate Oxidase