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Literature summary extracted from

  • Kataoka, K.; Kitagawa, R.; Inoue, M.; Naruse, D.; Sakurai, T.; Huang, H.W.
    Point mutations at the type I Cu ligands, Cys457 and Met467, and at the putative proton donor, Asp105, in Myrothecium verrucaria bilirubin oxidase and reactions with dioxygen (2005), Biochemistry, 44, 7004-7012.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.3.5 Pichia pastoris GS115 transformed using the pPICBO derivative linearized with Bpu1102I Albifimbria verrucaria

Protein Variants

EC Number Protein Variants Comment Organism
1.3.3.5 C457S can react with dioxygen, affords reaction intermediate I with absorption maxima at 340, 470, and 675 nm Albifimbria verrucaria
1.3.3.5 D105A exhibits 7.5% bilirubin oxidase activity compared to the wild-type enzyme, indicating that Asp105 conserved in all multi-copper oxidases donates a proton to reaction intermediates I and II Albifimbria verrucaria
1.3.3.5 D105E exhibits 46% bilirubin oxidase activity compared to the wild-type enzyme, indicating that Asp105 conserved in all multi-copper oxidases donates a proton to reaction intermediates I and II Albifimbria verrucaria
1.3.3.5 D105N does not react with dioxygen Albifimbria verrucaria
1.3.3.5 M467G with modified spectroscopic properties and redox potential, affords reaction intermediate II with absorption maxima at 355 and 450 nm Albifimbria verrucaria

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.3.3.5 66000
-
 all mutants expressed in Pichia pastoris, SDS-PAGE Albifimbria verrucaria

Organism

EC Number Organism UniProt Comment Textmining
1.3.3.5 Albifimbria verrucaria
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.3.5 by gel filtration Albifimbria verrucaria

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.3.3.5 additional information
-
 no bilirubin oxidase activity of mutant C457S and mutant D105N Albifimbria verrucaria
1.3.3.5 0.3
-
 ferrocyanide oxidase activity of mutant M467G Albifimbria verrucaria
1.3.3.5 1.2
-
 bilirubin oxidase activity of mutant D105A Albifimbria verrucaria
1.3.3.5 11
-
 bilirubin oxidase activity of mutant D105E Albifimbria verrucaria
1.3.3.5 24
-
 bilirubin oxidase activity of authentic enzyme Albifimbria verrucaria
1.3.3.5 24
-
 bilirubin oxidase activity of recombinant enzyme Albifimbria verrucaria

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.3.5 bilirubin + O2
-
 Albifimbria verrucaria biliverdin + H2O
-
 ?
1.3.3.5 ferrocyanide + O2
-
 Albifimbria verrucaria ?
-
 ?