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Information on EC 1.3.3.5 - bilirubin oxidase and Organism(s) Albifimbria verrucaria and UniProt Accession Q12737
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1.3.3.5 bilirubin oxidaseSpecify your search results
Word Map
- 1.3.3.5
-
electrode
- oxidases
-
biofuel
-
cathode
- ceruloplasmin
- myrothecium
-
anode
- laccases
- verrucaria
-
biocathode
- ferroxidase
-
trinuclear
- abts
-
laccase-like
- dioxygen
-
bioanode
-
electrocatalytic
-
bioelectrocatalytic
- 2,6-dimethoxyphenol
- biliverdin
-
cuprous
-
multi-copper
- syringaldazine
- copa
-
copper-binding
-
four-electron
-
mniii
-
self-powered
-
aceruloplasminemia
- hephaestin
- p-phenylenediamine
-
manganeseii
-
bioelectrodes
-
open-circuit
-
membrane-less
- cupredoxins
- analysis
-
electrocatalysts
-
manganese-oxidizing
-
methionine-rich
-
diazo
-
biodevice
- energy production
- medicine
- diagnostics
- biotechnology
- synthesis
- environmental protection
- industry
The taxonomic range for the selected organisms is: Albifimbria verrucaria
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
bilirubin oxidase, multicopper oxidase, copper oxidase, mvbod, blue cu enzyme, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
bilirubin oxidase
bilirubin oxidase M-1
-
-
-
-
BOD
-
-
oxidase, bilirubin
-
-
-
-
bilirubin oxidase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 bilirubin + O2 = 2 biliverdin + 2 H2O
analyis of the oxygen reduction reaction mechanism, detailed overview. Bilirubin oxidase (BOD) is the best catalyst for converting oxygen directly to water because of the very low overpotential necessary to catalyze the reaction
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
redox reaction
-
-
-
-
oxidation
reduction
oxidation
-
-
-
-
reduction
-
-
-
-
PATHWAY SOURCE
PATHWAYS
SYSTEMATIC NAME
IUBMB Comments
bilirubin:oxygen oxidoreductase
-
CAS REGISTRY NUMBER
COMMENTARY
80619-01-8
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + O2
?
-
-
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + O2
? + H2O
-
-
-
?
(4E,15Z)-cyclobilirubin IX alpha + O2
?
-
phosphate buffer, pH 3.5-7.4
-
-
?
(4Z,15E)-bilirubin IX alpha + O2
?
-
no activity above pH 4.5 in phosphate buffer
-
-
?
(4Z,15Z)-bilirubin IX alpha + O2
?
-
no activity above pH 5.5 in phosphate buffer, no acticvity in citrate-lactate buffer, pH 3.7
-
-
?
1,1'-dimethylferrocene + O2
1,1'-dimethylferricenium + H2O
-
1,1'-dimethylferrocene soluble as an inclusion complex with 2-hydroxypropyl-beta-cyclodextrin
-
?
2 bilirubin + O2
2 biliverdin + H2O
-
-
-
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + O2
? + H2O
-
BOD encapsulated in ananostructured sol-gel/carbon nanotube composite electrode effectively catalyzes the reduction of molecular oxygen into water through direct electron transfer
-
-
?
cytochrome c + O2
?
-
BOD efficiently accepts cytochrome c as an electron donor in both cases when cytochrome c is in solution or electrostatically adsorbed
-
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + O2
?
-
-
-
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + O2
?
-
i.e. ABTS
-
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + O2
?
-
the enzyme has a higher affinity for oxygen in the presence of the nanoparticles compared to the dissolved 2,2`-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) mediator, although with a slower turnover rate
-
-
?
bilirubin + O2
biliverdin + H2O
-
-
-
-
?
bilirubin + O2
biliverdin + H2O
-
enzyme also exhibits laccase activity
-
?
N,N-dimethyl-p-phenylenediamine + O2
?
-
23% of activity with bilirubin
-
-
?
additional information
?
-
direct proton uptake of Glu463 plays a key role in the proton donation to the activated oxygen species in the catalytic cycle, redox-induced protonation state changes of Glu463, electrochemistry-induced ATR-FTIR spectroscopy, overview
-
-
?
additional information
?
-
in addition to traditional laccase substrates like 2, 2'-azino-bis (3-ethylbenzthiazoline-6-sulfonate), syringaldazine, or 2,6-dimethoxyphenol, the enzyme catalyzes also the oxidation of conjugated and/or unconjugated bilirubin
-
-
?
additional information
?
-
oxidative polymerization of dihydroquercetin from Larix sibirica using bilirubin oxidase as a biocatalyst. DHQ oligomers (oligoDHQ) with molecular mass of 2800 and polydispersity of 8.6 are obtained by enzymatic reaction under optimal conditions. The oligomers appear to be soluble in dimethylsulfoxide, dimethylformamide, and methanol. UVvisible spectra of oligoDHQ in dimethylsulfoxide indicate the presence of highly conjugated bonds. Irregular structure of a polymer formed via the enzymatic oxidation of DHQ followed by nonselective radical polymerization. As compared with the monomer, oligoDHQ demonstrates higher thermal stability and high antioxidant activity
-
-
?
additional information
?
-
the enzyme needs to be fully reduced before it gets activated for catalysis
-
-
?
additional information
?
-
-
the enzyme needs to be fully reduced before it gets activated for catalysis
-
-
?
additional information
?
-
-
cytochrome c and bilirubin oxidase are coimmobilized in a polyelectrolyte multilayer on gold electrodes and although these two proteins are not natural reaction partners, the protein architecture facilitates an electron transfer from the electrode through multiple protein layers to molecular oxygen
-
-
?
additional information
?
-
-
activity measurement with Fe(CN)6 4-
-
-
?
additional information
?
-
-
decolorization and biodegradation of remazol brilliant blue R, an anthraquinone dye, by bilirubin oxidase
-
-
?
additional information
?
-
-
extracellular bilirubin oxidase decolorizes indigo carmine, biosorption and biodegradation of the dye is achved with more than 98% decolorization efficiency after 7 days at 26°C. Additionally, the crude bilirubin oxidase can efficiently decolorize indigo carmine at 30°C to 50°C and pH 5.5-9.5 with dye concentrations of 50-200 mg/ml, overview
-
-
?
additional information
?
-
-
the enzyme is also active with the laccase substrate 2, 2'-azino-bis (3-ethylbenzthiazoline-6-sulfonate)
-
-
?
additional information
?
-
-
the oxygen reduction reaction activity of the enzyme with direct electron transfer reactions is higher than that with mediated electron transfer reactions
-
-
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2 bilirubin + O2
2 biliverdin + H2O
-
-
-
-
?
additional information
?
-
in addition to traditional laccase substrates like 2, 2'-azino-bis (3-ethylbenzthiazoline-6-sulfonate), syringaldazine, or 2,6-dimethoxyphenol, the enzyme catalyzes also the oxidation of conjugated and/or unconjugated bilirubin
-
-
?
additional information
?
-
-
extracellular bilirubin oxidase decolorizes indigo carmine, biosorption and biodegradation of the dye is achved with more than 98% decolorization efficiency after 7 days at 26°C. Additionally, the crude bilirubin oxidase can efficiently decolorize indigo carmine at 30°C to 50°C and pH 5.5-9.5 with dye concentrations of 50-200 mg/ml, overview
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
ferrocyanide is an effective electron donor to type 1 Cu2+
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
copper
Cu
the enzyme consists of 3 cupredoxin-like domains with 4 copper ions forming two active sites
Cu2+
copper
Cu
Cu2+
additional information
BODs display a high tolerance towards chloride anions and other chelators
Cu2+
multicopper oxidase with copper-binding sites. The type I Cu is coordinated by four amino-acid residues His398, Cys457, His462 and Met 467, and the type II Cu is coordinated by two amino-acid residues, His94 and His401, binding structure, overview
Cu2+
multicopper oxidase, ferrocyanide is an effective electron donor to type 1 Cu2+
Cu2+
bilirubin oxidase utilizes four Cu+/2+ ions to reduce O2 to water. It contains four histidine-rich copper-binding domains
copper
-
Cu2+ can be removed by KCN, reconstitution of the apoprotein by treating with CuSO4 for 40 h at 4°C and pH 7-8, Cu2+ reconstituted enzyme regains full activity, Fe2+ reconstituts 59,1% activity, Co2+ reconstituts 31% activity and Cd2+ reconstituts 24.5% activity
copper
-
multicopper enzyme, contains type 1,2 and 3 copper, authentic and recombinant wild-type enzyme contain 4 copper atoms/protein
copper
-
multicopper oxidase contains 4 copper ions per protein molecule. Cu-binding sites are not affected by differences in carbohydrate content and the N-terminal extension of four amino acid residues
copper
-
multicopper oxidase, the redox state of type I Cu is an equilibrium state of the oxidized and reduced forms highly depending on pH, possibly by a shift of the radical center between Cu and cys sulfur
Cu
-
the enzyme contains type I copper and a trinuclear copper center comprised of a type II copper and a pair of type III coppers
Cu
-
the enzyme possesses a trinuclear Cu center, which is composed of one T2Cu and a pair of T3Cu atoms, T3aCu and T3bCu
Cu2+
-
multicopper oxidase containing four Cu centers, type 1 Cu, type II Cu, and a pair of type III Cu's in a protein molecule consisting of three domains with homologous structure to cupredoxin containing only type I Cu
Cu2+
-
dissociation of type I copper, caused by thermal inactivation, is accompanied by a conformational change and a decrease in secondary structure
Cu2+
-
highly activating, the enzyme contains four copper ions per functional unit
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
bilirubin
-
0.09 mM, complete inhibition of enzyme activity in 50 : 50 chloroform-n-heptane two-phase system
guanidinium hydrochloride
-
reversible inactivation at 1 M, pH 7.0, kinetics, overview
additional information
BODs display a high tolerance towards chloride anions and other chelators
-
additional information
-
measurements of reversible and irreversible inactivation processes of the redox enzyme bilirubin oxidase by electrochemical methods based on bioelectrocatalysis, overview
-
additional information
-
the enzyme is not affected by urea and EDTA
-
additional information
-
not affected by ascorbic acid, urea, D-glucose, uric acid, or triglycerides
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
-
i.e. ABTS, can serve as an electron mediator to facilitate the oxidation of remazol brilliant blue R
additional information
the enzyme needs to be fully reduced before it gets activated for catalysis
-
additional information
-
the enzyme needs to be fully reduced before it gets activated for catalysis
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
20.1
phenol 2,6-dimethoxyphenol
wild type enzyme, at pH 6.8 and 27°C
-
0.076
2,2'-azino-bis(3-ethylbenzthioline-6-sulfonic acid)
-
at pH 2.7, Km increases with pH
additional information
additional information
-
first-order kinetics, voltammogram for the bioelectrolytic reaction, overview
-
0.3
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
wild type enzyme, at pH 4.0 and 27°C
3.1
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
mutant enzyme W396A, at pH 4.0 and 27°C
6.8
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
mutant enzyme W396F, at pH 4.0 and 27°C
0.12
bilirubin
conjugated bilirubin, pH 5.5, temperature not specified in the publication
0.37
bilirubin
unconjugated bilirubin, pH 8.0, temperature not specified in the publication
0.25
2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)
-
pH 6.5, authentic bilirubin oxidase
0.34
2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)
-
pH 6.5, recombinant enzyme from Pichia pastoris
0.0602
bilirubin
-
preparations of enzyme lyophilized from N-(2-hydroxyethyl)-piperazine-N'-(3-propane-sulfonic acid) buffer suspended in 50 : 50 v/v chloroform-n-heptane mixture
0.14
ditaurobilirubin
-
pH 5.5, recombinant enzyme from Pichia pastoris
9.6
p-phenylenediamine
-
pH 6.5, recombinant enzyme from Pichia pastoris
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
7.17
2,2'-azino-bis(3-ethylbenzthioline-6-sulfonic acid)
-
at pH 5.3, about 3fold higher value at pH 2.5
94.3
bilirubin
unconjugated bilirubin, pH 8.0, temperature not specified in the publication
117
bilirubin
conjugated bilirubin, pH 5.5, temperature not specified in the publication
115
2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)
-
pH 6.5, authentic bilirubin oxidase
164
2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)
-
pH 6.5, recombinant enzyme from Pichia pastoris
0.00133
bilirubin
-
preparations of enzyme lyophilized from N-(2-hydroxyethyl)-piperazine-N'-(3-propane-sulfonic acid) buffer suspended in 50 : 50 v/v chloroform-n-heptane mixture
233
ditaurobilirubin
-
pH 5.5, recombinant enzyme from Pichia pastoris
90
p-phenylenediamine
-
pH 6.5, recombinant enzyme from Pichia pastoris
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2500
bilirubin
unconjugated bilirubin, pH 8.0, temperature not specified in the publication
9800
bilirubin
conjugated bilirubin, pH 5.5, temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
2.83
214.3
-
purified enzyme after anion exchange chromatography step, pH 7.5, 25°C
24
25.4
-
purified enzyme with substrate 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), pH 6.0, 24°C
additional information
2.83
-
commercial preparation, pH not specified in the publication, temperature not specified in the publication
additional information
-
3.3 units/ml, 1 unit is defined as the equivalent amount of enzyme that reduces DELTA A at 440 nm by 1.05/min, enzyme activity in crude extracts
additional information
-
206 units/A280, 1 unit is defined as the equivalent amount of enzyme that reduces DELTA A at 440 nm by 1.05/min, enzyme activity in crude extracts
additional information
-
no bilirubin oxidase activity of mutant C457S and mutant D105N
additional information
-
relative activity 0.09, 0.2 g/l BOD stored in 0.5 g/l poly[ethyleneimine], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.09, 0.2 g/l BOD stored in 2 g/l poly[ethyleneimine], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.17, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M H2PO4--HPO42- buffer system, no additive
additional information
-
relative activity 0.17, 0.2 g/l BOD stored in 0.1 M H2PO4--HPO42- buffer system, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.19, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M MOPS0-MOPS- buffer system, no additive
additional information
-
relative activity 0.22, 0.2 g/l BOD stored in 0.5 g/l poly[(dimethylimino)(2-oxo-1,2-ethanediyl)imino-1,6-hexanediylimino(1-oxo-1,2-ethanediyl)(dimethylimino)-1,6-hexanediyl], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.22, 0.2 g/l BOD stored in 0.5 g/l poly[(dimethylimino)-1,6-hexanediyl], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.22, 0.2 g/l BOD stored in 2 g/l poly[(dimethylimino)(2-oxo-1,2-ethanediyl)imino-1,6-hexanediylimino(1-oxo-1,2-ethanediyl)(dimethylimino)-1,6-hexanediyl], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.22, 0.2 g/l BOD stored in 2 g/l poly[(dimethylimino)-1,6-hexanediyl], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.23, 0.2 g/l BOD stored in 0.5 g/l poly(diallydimethylammonium), 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.23, 0.2 g/l BOD stored in 2 g/l poly(diallydimethylammonium), 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.25, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M H2PO4--HPO42- buffer system, 0.5 g/l poly[oxyethylene(dimethylimino)propyl(dimethylimino)ethylene]
additional information
-
relative activity 0.26, 0.2 g/l BOD stored in 0.5 g/l poly[metacrylpropyltrimethylammonium], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.26, 0.2 g/l BOD stored in 2 g/l poly[metacrylpropyltrimethylammonium], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.27, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M H2PO4--HPO42- buffer system, 0.5 g/l poly[(dimethylimino)(2-oxo-1,2-ethanediyl)imino-1,2-ethanediylimino(1-oxo-1,2-ethanediyl)-(dimethylimino)-1,3-propanediyl]
additional information
-
relative activity 0.27, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M H2PO4--HPO42- buffer system, 0.5 g/l poly[bis(2-chloroethyl)ether-alt-1,3-bis[3-(dimethylimino)propyl]urea]
additional information
-
relative activity 0.27, 0.2 g/l BOD stored in 0.5 g/l poly[(dimethylimino)(2-oxo-1,2-ethanediyl)imino-1,2-ethanediylimino(1-oxo-1,2-ethanediyl)(dimethylimino)-1,3-propanediyl], 0.1 M H2PO4--HPO42- buffer system, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.27, 0.2 g/l BOD stored in 0.5 g/l poly[bis(2-chloroethyl)ether-alt-1,3-bis[3-(dimethylimino)propyl]urea], 0.1 M H2PO4--HPO42- buffer system, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.28, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M H2PO4--HPO42- buffer system, 0.5 g/l poly(dimethylamine-co-epichrohydrin)
additional information
-
relative activity 0.28, 0.2 g/l BOD stored in 0.5 g/l poly(dimethylamine-co-epichrohydrin), 0.1 M H2PO4--HPO42- buffer system, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.28, 0.2 g/l BOD stored in 0.5 g/l poly[allyamine], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.28, 0.2 g/l BOD stored in 2 g/l poly[allyamine], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.30, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M MOPS0-MOPS- buffer system, 0.5 g/l poly[bis(2-chloroethyl)ether-alt-1,3-bis[3-(dimethylimino)propyl]urea]
additional information
-
relative activity 0.30, 0.2 g/l BOD stored in 0.5 g/l poly[bis(2-chloroethyl)ether-alt-1,3-bis[3-(dimethylimino)propyl]urea], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.30, 0.2 g/l BOD stored in 2 g/l poly[bis(2-chloroethyl)ether-alt-1,3-bis[3-(dimethylimino)propyl]urea], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.31, 0.2 g/l BOD stored in 0.5 g/l poly[lysine], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.31, 0.2 g/l BOD stored in 2 g/l poly[lysine], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.38, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M MOPS0-MOPS- buffer system, 0.5 g/l poly(dimethylamine-co-epichrohydrin)
additional information
-
relative activity 0.38, 0.2 g/l BOD stored in 0.5 g/l poly(dimethylamine-co-epichrohydrin), 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.38, 0.2 g/l BOD stored in 2 g/l poly(dimethylamine-co-epichrohydrin), 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.40, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M MOPS0-MOPS- buffer system, 0.5 g/l poly[oxyethylene(dimethylimino)propyl(dimethylimino)ethylene]
additional information
-
relative activity 0.43, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M MOPS0-MOPS- buffer system, 0.5 g/l poly[(dimethylimino)(2-oxo-1,2-ethanediyl)imino-1,2-ethanediylimino(1-oxo-1,2-etanediyl)-(dimethylimino)-1,3-propanediyl]
additional information
-
relative activity 0.43, 0.2 g/l BOD stored in 0.5 g/l poly[(dimethylimino)(2-oxo-1,2-ethanediyl)imino-1,2-ethanediylimino(1-oxo-1,2-ethanediyl)(dimethylimino)-1,3-propanediyl], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.43, 0.2 g/l BOD stored in 2 g/l poly[(dimethylimino)(2-oxo-1,2-ethanediyl)imino-1,2-ethanediylimino(1-oxo-1,2-ethanediyl)(dimethylimino)-1,3-propanediyl], 0.1 M MOPS-NaOH solution, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.51, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M BisTrisH+-BisTris0 buffer system, no additive
additional information
-
relative activity 0.51, 0.2 g/l BOD stored in 0.1 M BisTrisH+-BisTris0 buffer system, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.58, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M BisTrisH+-BisTris0 buffer system, 0.5 g/l poly[(dimethylimino)(2-oxo-1,2-ethanediyl)imino-1,2-ethanediylimino(1-oxo-1,2-etanediyl)-(dimethylimino)-1,3-propanediyl]
additional information
-
relative activity 0.58, 0.2 g/l BOD stored in 0.5 g/l poly[(dimethylimino)(2-oxo-1,2-ethanediyl)imino-1,2-ethanediylimino(1-oxo-1,2-ethanediyl)(dimethylimino)-1,3-propanediyl], 0.1 M BisTrisH+-BisTris0 buffer system, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.64, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M BisTrisH+-BisTris0 buffer system, 0.5 g/l poly[bis(2-chloroethyl)ether-alt-1,3-bis[3-(dimethylimino)propyl]urea]
additional information
-
relative activity 0.64, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M BisTrisH+-BisTris0 buffer system, 0.5 g/l poly[oxyethylene(dimethylimino)propyl(dimethylimino)ethylene]
additional information
-
relative activity 0.64, 0.2 g/l BOD stored in 0.5 g/l poly[bis(2-chloroethyl)ether-alt-1,3-bis[3-(dimethylimino)propyl]urea], 0.1 M BisTrisH+-BisTris0 buffer system, pH 7.0, 30°C, 2 days
additional information
-
relative activity 0.89, 0.2 g/l BOD stored at 30°C for 2 days in 0.1 M BisTrisH+-BisTris0 buffer system, 0.5 g/l poly(dimethylamine-co-epichrohydrin)
additional information
-
relative activity 0.89, 0.2 g/l BOD stored in 0.5 g/l poly(dimethylamine-co-epichrohydrin), 0.1 M BisTrisH+-BisTris0 buffer system, pH 7.0, 30°C, 2 days
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
2.7
-
oxidation of 2,2'-azino-bis(3-ethylbenzthioline-6-sulfonic acid)
4
-
using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) as a substrate
5
-
with substrate remazol brilliant blue R, in absence of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
5 - 7
-
oxidation of 1,1'-dimethylferrocene, activity decreases above pH 7.5
7.5
8.2
additional information
BODs display a high activity and stability at neutral pH
6
-
assay at, substrate 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
8
-
with substrate remazol brilliant blue R and activating 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
3 - 9.5
activity range, inactive above and below. In reaction with electron donor substrates, the enzyme exhibits the maximal activity at acidic pH values: pH 4.0 for 2,2'-azino-bis-[3-ethylbenzthiazoline-6-sulfonic acid] (ABTS) and pH 3.0 for potassium ferricyanide. Catalytic activity decreases on pH increase, and the enzyme becomes completely inactive at pH above 9.5. At neutral pH values, bilirubin oxidase retains about 50% maximal activity in oxidation of both substrates. In reaction with a hydrogen atom donor (catechol), the pH profile of the enzyme activity is shifted to alkaline values: enzymatic activity is not exhibited at pH below 6.0. This is probably related with the higher reactivity of the substrate as a phenolate anion
4 - 9
analysis of pH dependence of the enzyme immobilized on electrodes, detailed overview
3.5 - 4.5
-
using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) as a substrate
4.5 - 8.2
-
under acidic condition enzyme oxidizes only conjugated bilirubin
5 - 8.5
-
activity range with substrate remazol brilliant blue R, profile overview
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
24
-
assay at, substrate 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
40
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
20 - 60
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
physiological function
additional information
evolution
bilirubin oxidase (BOD) is a sub-group of multicopper oxidases (MCOs) also utilizing four Cu+/2+ ions
evolution
the enzyme belongs to the multicopper oxidase (MCO) family. These enzymes have four copper atoms that are classified into three types according to their spectroscopic and magnetic properties: type I (T1), type II (T2) and type III (T3) Cu. The MCO family can be separated into two types by substrate specificity. The first group catalyzes the outer sphere oxidation of small organic substrates and include the plant and fungal laccases and ascorbate oxidase, CotA, bilirubin oxidase, and phenoxazinone synthase
physiological function
bilirubin oxidase (BOD) is one of the multicopper oxidoreductases family with many interesting possible biotechnological applications, including biocatalytic reduction of oxygen (O2) at natural pH, biosensing, biobleaching, bioremediation, chemical synthesis, and wine stabilization
physiological function
the enzyme is a copper-containing oxidase that catalyzes oxidation of various organic compounds, including phenolic ones, by molecular oxygen, the latter is reduced to water via a fourelectron mechanism
additional information
BODs have a high efficiency of decolorizing compounds such as Trypan blue and Remazol brilliant blue R under mild pH conditions
additional information
-
dissociation of type I copper, caused by thermal inactivation, is accompanied by a conformational change and a decrease in secondary structure
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). BLRO_ALBVE
572
0
63948
Swiss-Prot
Secretory Pathway (Reliability: 2)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
63000
64000
-
1 * 64000, difference to deduced MW may be due to glycosylation, SDS-PAGE
66000
68000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
monomer
additional information
monomer
-
1 * 64000, difference to deduced MW may be due to glycosylation, SDS-PAGE
additional information
molecular-dynamics simulation of a BOD-mediator complex, mediator is 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), i.e. ABTS, overview
additional information
-
molecular-dynamics simulation of a BOD-mediator complex, mediator is 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), i.e. ABTS, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
glycoprotein
glycoprotein
-
two putative N-glycosylation sites at positions 472 and 482
glycoprotein
-
the recombinant enzyme expressed in Pichia pastoris contains non-native type N-linked sugar chains
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method, 0.003 ml of 9.2 mg/ml protein in 10 mM Tris-HCl, pH 8.0, are mixed with 0.003 ml of recipitant solution containing 10% 2-methyl-2,4-pentanediol, 1.44 M ammonium sulfate, 10% glycerol and 0.5 M KCl, equilibration against 0.2-1.0 ml precipitant solution at 20°C, X-ray diffraction structure determination and analysis at 2.3-2.5 A resolution, modelling
wild type and mutant enzymes W396A and W396F bound to ferricyanide, hanging drop vapor diffusion method, using 0.1 M succinic acid, 14% (w/v) polyethylene glycol 3350
sitting drop vapor diffusion method, wild type enzyme is crystallized with 0.1 M sodium citrate pH 5.0 and 20% (w/v) PEG 8000. Mutant enzyme M47Q is crystallized with 0.1 M MES pH 5.5,12% (w/v) PEG 8000, and 0.1 M calcium acetate
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
W396A
the mutant shows increased activity with bilirubin, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) or ferricyanide compared to the wild type enzyme
W396D
the mutant shows practically zero activity compared to the wild type enzyme
W396F
the mutant shows decreased activity with bilirubin and increased activity with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) or ferricyanide compared to the wild type enzyme
biotechnology
-
bilirubin oxidase has been found to be the best enzyme for converting O2 to H2O as a cathodic enzyme in biofuel cells
C457S
-
can react with dioxygen, affords reaction intermediate I with absorption maxima at 340, 470, and 675 nm
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
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
H456D/H458D
-
mutant with weak bilirubin oxidase and ferroxidase activity
H456K/H458K
-
mutant with weak bilirubin oxidase and ferroxidase activity
M467F
-
the mutated type I Cu center shows characteristics of phytocyanins
M467G
M467L
-
the mutated type I Cu center shows characteristics of phytocyanins
M467Q
N394A/W396T
-
the enzymatic activity of the mutant is prominently decreased compared to the wild type enzyme. The enzyme shows shifts in the redox potential of type I copper towards negative direction by more than 100 mV and decreases in cathodic current in electrochemistry, whereas optical and magnetic properties of type I copper are not affected or sparingly affected
N459A/M467F
-
activity is decreased to 1% of the recombinant wild type enzyme, the mutated type I Cu center shows characteristics of phytocyanins, blue copper proteins with an axial coordination of Gln, due to compensatory binding of the distal Asn459
W396A
-
the enzymatic activity of the mutant is prominently decreased compared to the wild type enzyme. The enzyme shows shifts in the redox potential of type I copper towards negative direction by more than 100 mV and decreases in cathodic current in electrochemistry, whereas optical and magnetic properties of type I copper are not affected or sparingly affected
W396F
-
the enzymatic activity of the mutant is prominently decreased compared to the wild type enzyme. The enzyme shows shifts in the redox potential of type I copper towards negative direction by more than 100 mV and decreases in cathodic current in electrochemistry, whereas optical and magnetic properties of type I copper are not affected or sparingly affected
W396T
-
the enzymatic activity of the mutant is prominently decreased compared to the wild type enzyme. The enzyme shows shifts in the redox potential of type I copper towards negative direction by more than 100 mV and decreases in cathodic current in electrochemistry, whereas optical and magnetic properties of type I copper are not affected or sparingly affected
W396Y
-
the enzymatic activity of the mutant is prominently decreased compared to the wild type enzyme. The enzyme shows shifts in the redox potential of type I copper towards negative direction by more than 100 mV and decreases in cathodic current in electrochemistry, whereas optical and magnetic properties of type I copper are not affected or sparingly affected
additional information
M467G
-
with modified spectroscopic properties and redox potential, affords reaction intermediate II with absorption maxima at 355 and 450 nm
M467Q
-
the enzymatic activity of the mutant is very low toward bilirubin but it works as a good catalyst in direct electron transfer-type bioelectrocatalytic reduction of dioxygen into water, the kcat value is 3fold increased
M467Q
-
the catalytic activity of the mutant is quite low (about 0.3% of wild type activity)
M467Q
-
the mutant is inactive against bilirubin. A post-translational crosslink between Trp396 and His398, formed in the vicinity of the T1Cu site in wild type enzyme, is absent in the mutant
additional information
enzyme immobilization and preparation of a bilirubin oxidase-based air breathing cathode, evaluation by constant monitoring over 45 days, analysis of effect of electrolyte composition on the cathode oxygen reduction reaction output, and of deactivation of the electrocatalytic activity of the enzyme in phosphate buffer saline solution and in activated sludge, anions and cations in autoclaved activated sludge with PBS, overview. Enzyme deactivation is also studied in activated sludge to simulate an environment close to the real waste operation with pollutants, solid particles and bacteria. The presence of low-molecularweight soluble contaminants is identified as the main reason for an immediate enzymatic deactivation within few hours of cathode operation. The presence of solid particles and bacteria does not affect the natural degradation of the enzyme
additional information
enzyme immobilization on an electrode, bilirubin oxidase adsorbed on a nanocomposite modified electrode has three distinct redox sites within that show pH dependence: the 1st redox centre with the highest redox potential Ec(1st) = 404 mV vs. Ag/AgCl (614 mV vs. NHE at pH 7.0) exhibits pH dependence with a slope -dEc(1st)/dpH = 66 mVunder a non-turnover process. The 2nd redox centre with a potential Ec(2nd) = 228 mV vs. Ag/AgCl (438 mV vs. NHE at pH 7.0) is not dependent on pH in the absence and presence of O2. Finally, the 3rd redox site with a redox potential Ec(3rd) = 92 mVvs. Ag/AgCl (302 mV vs. NHE at pH 7.0) exhibits pH dependence for a cathodic process with -dEc(3rd)/dpH = 70 mV and for anodic process with -dEa(3rd)/dpH = 73 mV, respectively. Two break points for dependence of Ec(1st) or Ec(3rd) on pH are observed for the 1st (T1) site and the 3rd site assigned to involvement of two acidic amino acids, Asp105 and Glu463. Potential difference between cathodic peaks of BOD as a dependence on pH, overview
additional information
immobilization of bilirubin oxidase on graphene oxide flakes with different negative charge density for oxygen reduction, effect of graphene oxide charge density on enzyme coverage, electron transfer rate and current density. An effective bilirubin oxidase-based biocathode using graphene oxide can be prepared in 2 steps: 1. electrostatic adsorption of the enzyme on graphene oxide, 2. electrochemical reduction of the enzyme-elektrode composite to form a electrochemically reduced graphene oxide-enzyme film (BOD-ErGO) on the electrode, identification of an optimal charge density of graphene oxide for BOD-ErGO composite preparation, method evaluation and electrode characterization, detailed overview. Results reveal that 1. there is an optimal density of a negative surface charge density needed to obtain high j, GAMMA and kS, 2. a lower negative surface charge density is needed to achieve the highest kS compared to j, GAMMA, and 3. current density is influenced mainly by GAMMA and to lesser extent by kS, suggesting that the electron exchange in all cases is fast enough for not being a limiting factor in a biocatalytic current generation
additional information
mediator-free direct electron transfer (DET)-type of immobilization between biocatalysts and electrode surface is achieved by favorable adsorption of BOD on solid surfaces: electrochemical conditioning of aminocarbon nanotubes on a graphene support in an alkaline solution is used to produce -NHOH as hydrophilic functional groups for the efficient immobilization of bilirubin oxidase enzyme. Application of the immobilized enzyme for direct electrocatalytic reduction of O2 is investigated. The onset potential of 0.81 V versus NHE and peak current density of 2.3 mAcm2 for rotating modified electrode at 1250 rpm indicate improved biocatalytic activity of the proposed system for O2 reduction, immobilization of the enzyme on target amino-CNT-Gr modified electrode, method evaluation, detailed overview
additional information
mediator-less, direct electro-catalytic reduction of oxygen to water is achieved on spectrographite electrodes modified by physical adsorption of bilirubin oxidases from Myrothecium verrucaria. The existence of an alternative resting form of the enzyme is validated. Effects on the catalytic cycle by temperature, pH and the presence of halogens in the buffer, overview
additional information
-
mediator-less, direct electro-catalytic reduction of oxygen to water is achieved on spectrographite electrodes modified by physical adsorption of bilirubin oxidases from Myrothecium verrucaria. The existence of an alternative resting form of the enzyme is validated. Effects on the catalytic cycle by temperature, pH and the presence of halogens in the buffer, overview
additional information
-
attachment of the purified enzyme to a PGE surface, from pyrolytic graphite plates, modified with 6-amino-2-naphthoic acid, modifiers tested in increasing activity are 4-aminothiophenol, 4-aminobenzonitrile, 2-aminoacridine, 2-aminoanthracene, 1-aminoanthracene, 2-aminochrysene, 4-benzyloxyaniline, 4-aminobenzoic acid, 4-aminophenylacetic acid, 6-amino-2-naphthoic acid, the latter giving best results. Method optimization, cyclic voltammetry measuring the electrocatalytic reduction of dioxygen by platinum electrodeposited on PGE, detailed overview
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8
-
stability drops when 100 mM cetyltrimethyl ammonium bromide are added to the aqueous solution, half-life of inactivation: 1500 min
391027
9.2 - 9.7
additional information
additional information
BODs display a high activity and stability at neutral pH
724076
additional information
-
the reaction of BOD at gold electrodes is shown to be efficient only at low pH
686371
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
60
20 - 60
37
40 - 50
-
the enzyme activity is stable after incubation for 1 h at a temperature up to 40°C, thermal stability is decreased beyond 50?C
40 - 61
-
sol-gel encapsulated BOD shows no decrease in relative activity until over 50°C while free enzyme loses some 40% activity at 40°C
50 - 70
-
the half life of BOD at 50°C is 114 min, residual activity after 30 min at 50°C is 83.3% of the original activity, at 70°C BOD activity disappears within several min
60
70
60
half-life of the wild-type enzyme is 15 min, and of the recombinant enzyme 90 min
20 - 60
-
1 h incubation at 60°C, free enzyme retains 26% of its original activity, immobilized enzyme 73%
37
-
soluble enzyme, half-life: 12 h immobilized enzyme, half-life: 63 h
60
-
half-life of the recombinant enzyme expressed in Escherichia coli is 90 min, half-life of authentic enzyme is 15 min
60
-
irreversible thermal inactivation, dynamics and kinetics, overview
70
-
lyophilized enzyme stored at 70°C in the presence of polyvinylalcohol for 5 h, 40% of acitivity are lost after rehydration
70
-
lyophilized enzyme stored at 70°C in the presence of alpha,beta-poly(N-hydroxyethyl)-L-aspartamide for 5 h, about 10% of acitivity are lost after rehydration
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
BOD can undergo non-catalytic direct electron transfer between enzyme and carbon electrode and retains its catalytic activity, 1%-2% of the immobilized BOD (as a first monolayer on the electrode surface) participates in direct electrical communication with a carbon electrode
-
multi-walled carbon nanotube modification of glassy carbon electrodes strongly enhances the oxygen reduction of bilirubin oxidase compared to unmodified electrodes, the quasi-reversible redox reaction might be attributed to the trinuclear T2/T3 cluster of BOD, a BOD-multi-walled carbon nanotube-modified electrode retains 70% of the initial activity for oxygen reduction after 4 days
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
chloroform
n-heptane
chloroform
-
water-in-oil cetyltrimethyl ammonium bromide microemulsions of the enzyme, 60% of activity decay with a rate constant of 0.0183/min, 40% are lost at a rate constant of 0.000278/min
chloroform
-
chloroform-n-heptane mixture, preparations of enzyme lyophilized from N-(2-hydroxyethyl)-piperazine-N'-(3-propane-sulfonic acid) buffer suspended in 50:50 v/v chloroform-n-heptane mixture are activ if more than 0.081% v/v water are present in the two-phase system
n-heptane
-
water-in-oil cetyltrimethyl ammonium bromide microemulsions of the enzyme, 60% of activity decay with a rate constant of 0.0183/min, 40% are lost at a rate constant of 0.000278/min
n-heptane
-
chloroform-n-heptane mixture, preparations of enzyme lyophilized from N-(2-hydroxyethyl)-piperazine-N'-(3-propane-sulfonic acid) buffer suspended in 50:50 v/v chloroform-n-heptane mixture are active if more than 0.081% v/v water are present in the two-phase system
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, 50 mM phosphate buffer, pH 7.4, less than 10% activity are lost after 2 days
-
4°C, enzyme nanoparticles bound to polyethylene film, reutilized 150times, in Na3PO4 buffer pH 7.0, 120 days, 50% loss of activity
-
freeze-dried, rehydrated at 15°C in the presence of trehalose, 20% loss of activity
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
the commercial preparation of bilirubin oxidase from Myrothecium verrucaria is additionally purified by anion exchange chromatography
ammonium sulfate precipitation, DEAE-cellulose column chromatography, and Sephadex-G-100 gel filtration
-
extracellular enzyme 3.92fold by ammonium sulfate fractionation and anion exchange chromatography, an additional step of gel filtration lowers the enzyme purity and amount
-
native enzyme by ammonium sulfate fractionation, anion exchange chromatography, and gel filtration
-
native enzyme by anion exchange chromatography and hydrophobic interaction chromatography
-
Toyopearl Butyl-650M column chromatography and Superdex 200 gel filtration
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Aspergillus oryzae, enzyme is in a resting form different from that of authentic bilirubin oxidase, but reaches the resting form of the authentic enzyme after one cycle of reduction and reoxidation with dioxygen
-
expression in Pichia pastoris. The cDNA encoding bilirubin oxidase is cloned into the Pichia pastoris expression vector pPIC9K under the control of the alcohol oxidase 1 promoter and its protein product is secreted using the Saccharomyces alpha-mating factor signal sequence
-
expression of H94V, H134/136V, C457V, C457A and H456/458V mutants in Aspergillus oryzae
-
Pichia pastoris GS115 transformed using the pPICBO derivative linearized with Bpu1102I
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
diagnostics
BOD is used for diagnostic analysis of bilirubin in serum
energy production
design of a bioanode that directly utilizes starch as a fuel in an enzymatic biofuel cell. The enzymatic fuel cell is based on three enzymes (alpha-amylase, glucoamylase and glucose oxidase). The carbon paste electrode containing these three enzymes and tetrathiafulvalene can both saccharize and oxidize starchy biomass. In cyclic voltammetry, catalytic currents are successfully observed with both glucose and starchy white rice used as a substrate. A membraneless white rice/O2 biofuel cell is assembled and the electrochemical performance is evaluated. The three enzyme based electrode is used as a bioanode and an immobilized bilirubin oxidase (derived from Myrothecium verrucaria) electrode is used as a biocathode. The biofuel cell delivers an open circuit voltage of 0.522 V and power density of up to 0.099 mW/cm
synthesis
analysis
biofuel production
-
the enzyme is used in a biofuel cell comprising enzymeless anode consisting of 0.2 mM braided gold wire modified with colloidal platinum and bilirubin oxidase-modified gold-coated Buckypaper biocathode
environmental protection
-
BOX can be used to decolorize synthetic dyes from effluents, especially for anthraquinonic dyes
industry
-
BOX can be used to decolorize synthetic dyes from effluents, especially for anthraquinonic dyes
medicine
additional information
analysis
BOD has attracted considerable attention as an enzymatic catalyst for the cathode of biofuel cells that work under neutral conditions
analysis
the ability to perform the oxygen reduction reaction under physiological conditions is the most interesting feature of the enzyme bilirubin oxidase in terms of the design of biocathodes for biofuel cell applications. The enzyme is commonly immobilized on a solid carrier to achieve the highest biocatalytic activity, stability, selectivity, and reusability. Optimization of the application of immobilized enzyme for direct electrocatalytic reduction of O2
synthesis
enzymatic oxidative polymerization of dihydroquercetin from Larix sibirica using bilirubin oxidase as a biocatalyst. As compared with the monomer, oligoDHQ demonstrates higher thermal stability and high antioxidant activity
synthesis
mediator-less, direct electro-catalytic reduction of oxygen to water is achieved on spectrographite electrodes modified by physical adsorption of bilirubin oxidases from Myrothecium verrucaria. Bilirubin oxidase (BOD) is the best catalyst for converting oxygen directly to water because of the very low overpotential necessary to catalyze the reaction
analysis
-
enzyme activity in serum bilirubin assays can be monitored by capillary electrophoresis
analysis
-
immobilized onto aminopropyl and arylamine glass beads to form an enzyme bioreactor
analysis
-
sandwich-type enzyme-amplified amperometric detection of DNA with ambient O2 as the substrate at neutral pH in the presence of chloride
analysis
-
application of polyammonium cations to enzyme-immobilized electrode: application as enzyme stabilizer for bilirubin oxidase
analysis
-
application of poly[oxyethylene(dimethylimino)propyl-(dimethylimino)ethylene] as enzyme stabilizer for bilirubin oxidase immobilized electrode
medicine
-
immobilized bilirubin oxidase reactor could be used for the degradation of bilirubin in neonatal jaundice
additional information
redox potential of the T1 site of BOD is close to 670 mV vs. NHE, redox potential of the T2 site is near 390 mV vs. NHE
additional information
-
redox potential of the T1 site of BOD is close to 670 mV vs. NHE, redox potential of the T2 site is near 390 mV vs. NHE
additional information
BODs have a high efficiency of decolorizing compounds such as Trypan blue and Remazol brilliant blue R under mild pH conditions
additional information
usage of the enzyme for a bilirubin oxidase-based air breathing cathode, evaluation by constant monitoring over 45 days, analysis of effect of electrolyte composition on the cathode oxygen reduction reaction output, and of deactivation of the electrocatalytic activity of the enzyme in phosphate buffer saline solution and in activated sludge. BOx electrochemical response as a function of presence of pollutants, solids and bacteria, overview
additional information
-
the purified bilirubin oxidase in Myrothecium verrucaria strain has potential application in dye effluent decolorization. Extracellular bilirubin oxidase decolorizes indigo carmine, biosorption and biodegradation of the dye is achieved with more than 98% decolorization efficiency after 7 days at 26°C. Additionally, the crude bilirubin oxidase can efficiently decolorize indigo carmine at 30°C to 50°C and pH 5.5-9.5 with dye concentrations of 50-200 mg/ml
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Murao, S.; Tanaka, N.
Isolation and identification of a microorganism producing bilirubin oxidase
Agric. Biol. Chem.
46
2031-2034
1982
Albifimbria verrucaria, Striaticonidium cinctum, Paramyrothecium roridum
-
Tanaka, N.; Murao, S.
Purification and some properties of bilirubin oxidase of Myrotheticum verrucaria MT-1
Agric. Biol. Chem.
46
2499-2503
1982
Albifimbria verrucaria
-
Gotoh, Y.; Kondo, Y.; Kaji, H.; Takeda, A.; Samejima, T.
Characterization of copper atoms in bilirubin oxidase by spectroscopic analyses
J. Biochem.
106
621-626
1989
Albifimbria verrucaria
Sung, C.; Lavin, A.; Klibanov, A.M.; Langer, R.
An immobilized enzyme reactor for the detoxification of bilirubin
Biotechnol. Bioeng.
28
1531-1539
1986
Albifimbria verrucaria
Murao, S.; Tanaka, N.
A new enzyme "bilirubin oxidase" produced by Myrotheticum verrucaria MT-1
Agric. Biol. Chem.
45
2383-2384
1981
Albifimbria verrucaria
-
Tanaka, N.; Murao, S.
Difference between various copper-containing enzymes (Polyporus laccase, mushroom tyrosinase and cucumber ascorbate oxidase) and bilirubin oxidase
Agric. Biol. Chem.
47
1627-1628
1983
Albifimbria verrucaria, Albifimbria verrucaria MT-1
-
Skrika-Alexopoulos, E.; Freedman, R.B.
Factors affecting enzyme characteristics of bilirubin oxidase suspensions in organic solvents
Biotechnol. Bioeng.
41
887-893
1993
Albifimbria verrucaria
Skrika-Alexopoulos, E.; Muir, J.; Freedman, R.B.
Stability of bilirubin oxidase in organic solvent media: A comparative study on two low-water systems
Biotechnol. Bioeng.
41
894-899
1993
Albifimbria verrucaria
Koikeda, S.; Ando, K.; Kaji, H.; Inoue, T.; Murao, S.; Takeuchi, K.; Samejima, T.
Molecular cloning of the gene for bilirubin oxidase from Myrothecium verrucaria and its expression in yeast
J. Biol. Chem.
268
18801-18809
1993
Albifimbria verrucaria
Male, K.B.; Luong, J.H.T.
Characterization and kinetic studies of a novel dye prepared from the oxidation of a water-soluble 1,1'-dimethylferrocene-2-hydroxypropyl-.beta-cyclodextrin complex using immobilized bilirubin oxidase
Enzyme Microb. Technol.
16
425-431
1994
Albifimbria verrucaria
-
Samejima, T.; Wu, C.S.; Shiboya, K.; Kaji, H.; Koikeda, S.; Ando, K.; Yang, J.T.
Conformation of bilirubin oxidase in native and denatured states
J. Protein Chem.
13
307-313
1994
Albifimbria verrucaria
Xu, F.; Shin, W.; Brown, S.H.; Wahleithner, J.A.; Sundaram, U.M.; Solomon, E.I.
A study of a series of recombinant fungal laccases and bilirubin oxidase that exhibit significant differences in redox potential, substrate specificity, and stability
Biochim. Biophys. Acta
1292
303-311
1996
Albifimbria verrucaria
Nakai, Y.; Yoshioka, S.; Aso, Y.; Kojima, S.
Solid-state rehydration-induced recovery of bilirubin oxidase activity in lyophilized formulations reduced during freeze-drying
Chem. Pharm. Bull.
46
1031-1033
1998
Albifimbria verrucaria
-
Chen, J.P.; Wang, H.Y.
Improved properties of bilirubin oxidase by entrapment in alginate-silicate sol-gel matrix
Biotechnol. Tech.
12
851-853
1998
Albifimbria verrucaria
Zhou, X.M.; Liu, J.W.; Zou, X.; Chen, J.J.
Monitoring catalytic reaction of bilirubin oxidase and determination of bilirubin and bilirubin oxidase activity by capillary electrophoresis
Electrophoresis
20
1916-1920
1999
Albifimbria verrucaria
Shimizu, A.; Kwon, J.H.; Sasaki, T.; Satoh, T.; Sakurai, N.; Sakurai, T.; Yamaguchi, S.; Samejima, T.
Myrothecium verrucaria Bilirubin Oxidase and Its Mutants for Potential Copper Ligands
Biochemistry
38
3034-3042
1999
Albifimbria verrucaria
Yoshioka, S.; Aso, Y.; Kojima, S.; Tanimoto, T.
Effect of polymer excipients on the enzyme activity of lyophilized bilirubin oxidase and beta-galactosidase formulations
Chem. Pharm. Bull.
48
283-285
2000
Albifimbria verrucaria
Itoh, S.; Kusaka, T.; Imai, T.; Isobe, K.; Onishi, S.
Effects of bilirubin and its photoisomers on direct bilirubin measurement using bilirubin oxidase
Ann. Clin. Biochem.
37
452-456
2000
Albifimbria verrucaria
Kim, H.H.; Zhang, Y.; Heller, A.
Bilirubin oxidase label for an enzyme-linked affinity assay with O2 as substrate in a neutral pH NACL solution
Anal. Chem.
76
2411-2414
2004
Albifimbria verrucaria
Sakurai, T.; Zhan, L.; Fujita, T.; Kataoka, K.; Shimizu, A.; Samejima, T.; Yamaguchi, S.
Authentic and recombinant bilirubin oxidases are in different resting forms
Biosci. Biotechnol. Biochem.
67
1157-1159
2003
Albifimbria verrucaria
Zoppellaro, G.; Sakurai, N.; Kataoka, K.; Sakurai, T.
The reversible change in the redox state of type I Cu in Myrothecium verrucaria bilirubin oxidase depending on pH
Biosci. Biotechnol. Biochem.
68
1998-2000
2004
Albifimbria verrucaria
Shimizu, A.; Samejima, T.; Hirota, S.; Yamaguchi, S.; Sakurai, N.; Sakurai, T.
Type III Cu mutants of Myrothecium verrucaria bilirubin oxidase
J. Biochem.
133
767-772
2003
Albifimbria verrucaria
Kataoka, K.; Tanaka, K.; Sakai, Y.; Sakurai, T.
High-level expression of Myrothecium verrucaria bilirubin oxidase in Pichia pastoris, and its facile purification and characterization
Protein Expr. Purif.
41
77-83
2005
Albifimbria verrucaria
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
Biochemistry
44
7004-7012
2005
Albifimbria verrucaria
Christenson, A.; Shleev, S.; Mano, N.; Heller, A.; Gorton, L.
Redox potentials of the blue copper sites of bilirubin oxidases
Biochim. Biophys. Acta
1757
1634-1641
2006
Ganoderma tsunodae, Albifimbria verrucaria (Q12737), Albifimbria verrucaria
Otsuka, K.; Sugihara, T.; Tsujino, Y.; Osakai, T.; Tamiya, E.
Electrochemical consideration on the optimum pH of bilirubin oxidase
Anal. Biochem.
370
98-106
2007
Albifimbria verrucaria, Albifimbria verrucaria BO3
Ikeda, T.; Tatsumi, H.; Katano, H.; Wanibuchi, M.; Hibi, T.; Kajino, T.
A bioelectrocatalysis method for the kinetic measurement of thermal inactivation of a redox enzyme, bilirubin oxidase
Anal. Sci.
24
237-241
2008
Albifimbria verrucaria
Dronov, R.; Kurth, D.G.; Moehwald, H.; Scheller, F.W.; Lisdat, F.
Communication in a protein stack: electron transfer between cytochrome c and bilirubin oxidase within a polyelectrolyte multilayer
Angew. Chem.
47
3000-3003
2008
Albifimbria verrucaria
Kataoka, K.; Tsukamoto, K.; Kitagawa, R.; Ito, T.; Sakurai, T.
Compensatory binding of an asparagine residue to the coordination-unsaturated type I Cu center in bilirubin oxidase mutants
Biochem. Biophys. Res. Commun.
371
416-419
2008
Albifimbria verrucaria
Ivnitski, D.; Artyushkova, K.; Atanassov, P.
Surface characterization and direct electrochemistry of redox copper centers of bilirubin oxidase from fungi Myrothecium verrucaria
Bioelectrochemistry
74
101-110
2008
Albifimbria verrucaria
Sakurai, T.; Kataoka, K.
Basic and applied features of multicopper oxidases, CueO, bilirubin oxidase, and laccase
Chem. Rec.
7
220-229
2007
Albifimbria verrucaria
Dronov, R.; Kurth, D.G.; Scheller, F.W.; Lisdat, F.
Direct and cytochrome c mediated electrochemistry of bilirubin oxidase on gold
Electroanalysis
19
1642-1646
2007
Albifimbria verrucaria
-
Weigel, M.C.; Tritscher, E.; Lisdat, F.
Direct electrochemical conversion of bilirubin oxidase at carbon nanotube-modified glassy carbon electrodes
Electrochem. Commun.
9
689-693
2007
Albifimbria verrucaria
-
Kamitaka, Y.; Tsujimura, S.; Kataoka, K.; Sakurai, T.; Ikeda, T.; Kano, K.
Effects of axial ligand mutation of the type I copper site in bilirubin oxidase on direct electron transfer-type bioelectrocatalytic reduction of dioxygen
J. Electroanal. Chem.
601
119-124
2007
Albifimbria verrucaria
-
Lim, J.; Cirigliano, N.; Wang, J.; Dunn, B.
Direct electron transfer in nanostructured sol-gel electrodes containing bilirubin oxidase
Phys. Chem. Chem. Phys.
9
1809-1814
2007
Albifimbria verrucaria
Katano, H.; Tatsumi, H.; Hibi, T.; Ikeda, T.; Tsukatani, T.
Application of polyammonium cations to enzyme-immobilized electrode: application as enzyme stabilizer for bilirubin oxidase
Anal. Sci.
24
1421-1424
2008
Albifimbria verrucaria
Katano, H.; Uematsu, K.; Hibi, T.; Ikeda, T.; Tsukatani, T.
Application of poly[oxyethylene(dimethylimino)propyl-(dimethylimino)ethylene] as enzyme stabilizer for bilirubin oxidase immobilized electrode
Anal. Sci.
25
1077-1081
2009
Albifimbria verrucaria
Mizutani, K.; Toyoda, M.; Sagara, K.; Takahashi, N.; Sato, A.; Kamitaka, Y.; Tsujimura, S.; Nakanishi, Y.; Sugiura, T.; Yamaguchi, S.; Kano, K.; Mikami, B.
X-ray analysis of bilirubin oxidase from Myrothecium verrucaria at 2.3 A resolution using a twinned crystal
Acta Crystallogr. Sect. F
66
765-770
2010
Albifimbria verrucaria (Q12737), Albifimbria verrucaria, Albifimbria verrucaria MT-1 (Q12737)
Ikeda, T.; Uematsu, K.; Ma, H.; Katano, H.; Hibi, T.
Measurements of reversible and irreversible inactivation processes of a redox enzyme, bilirubin oxidase, by electrochemical methods based on bioelectrocatalysis
Anal. Sci.
25
1283-1288
2009
Albifimbria verrucaria
Iwaki, M.; Kataoka, K.; Kajino, T.; Sugiyama, R.; Morishita, H.; Sakurai, T.
ATR-FTIR study of the protonation states of the Glu residue in the multicopper oxidases, CueO and bilirubin oxidase
FEBS Lett.
584
4027-4031
2010
Albifimbria verrucaria (Q12737)
Liu, Y.; Huang, J.; Zhang, X.
Decolorization and biodegradation of remazol brilliant blue R by bilirubin oxidase
J. Biosci. Bioeng.
108
496-500
2009
Albifimbria verrucaria, Albifimbria verrucaria IMER1
Dos Santos, L.; Climent, V.; Blanford, C.F.; Armstrong, F.A.
Mechanistic studies of the blue Cu enzyme, bilirubin oxidase, as a highly efficient electrocatalyst for the oxygen reduction reaction
Phys. Chem. Chem. Phys.
12
13962-13974
2010
Albifimbria verrucaria
Mano, N.
Features and applications of bilirubin oxidases
Appl. Microbiol. Biotechnol.
96
301-307
2012
Aspergillus sojae, Penicillium janthinellum, Pyricularia oryzae, Ganoderma tsunodae, Bacillus pumilus (A8FAG9), Bacillus subtilis (P07788), Albifimbria verrucaria (Q12737), Bacillus licheniformis (Q65MU7), Pleurotus ostreatus (Q9UVY4)
Han, X.; Zhao, M.; Lu, L.; Liu, Y.
Purification, characterization and decolorization of bilirubin oxidase from Myrothecium verrucaria 3.2190
Fungal Biol.
116
863-871
2012
Albifimbria verrucaria, Albifimbria verrucaria 3.2190
Khlupova, M.E.; Vasileva, I.S.; Shumakovich, G.P.; Morozova, O.V.; Chertkov, V.A.; Shestakova, A.K.; Kisin, A.V.; Yaropolov, A.I.
Enzymatic polymerization of dihydroquercetin using bilirubin oxidase
Biochemistry (Moscow)
80
233-241
2015
Albifimbria verrucaria (Q12737)
Santoro, C.; Babanova, S.; Erable, B.; Schuler, A.; Atanassov, P.
Bilirubin oxidase based enzymatic air-breathing cathode operation under pristine and contaminated conditions
Bioelectrochemistry
108
1-7
2016
Albifimbria verrucaria (Q12737)
Filip, J.; Tkac, J.
The pH dependence of the cathodic peak potential of the active sites in bilirubin oxidase
Bioelectrochemistry
96
14-20
2014
Albifimbria verrucaria (Q12737)
Filip, J.; Andicsova-Eckstein, A.; Vikartovska, A.; Tkac, J.
Immobilization of bilirubin oxidase on graphene oxide flakes with different negative charge density for oxygen reduction. The effect of GO charge density on enzyme coverage, electron transfer rate and current density
Biosens. Bioelectron.
89
384-389
2017
Albifimbria verrucaria (Q12737)
Navaee, A.; Salimi, A.; Jafari, F.
Electrochemical pretreatment of amino-carbon nanotubes on graphene support as a novel platform for bilirubin oxidase with improved bioelectrocatalytic activity towards oxygen reduction
Chemistry
21
4949-4953
2015
Albifimbria verrucaria (Q12737)
Tasca, F.; Farias, D.; Castro, C.; Acuna-Rougier, C.; Antiochia, R.
Bilirubin oxidase from Myrothecium verrucaria physically absorbed on graphite electrodes. Insights into the alternative resting form and the sources of activity loss
PLoS ONE
10
e0132181
2015
Albifimbria verrucaria (Q12737), Albifimbria verrucaria
Yamamoto, K.; Matsumoto, T.; Shimada, S.; Tanaka, T.; Kondo, A.
Starchy biomass-powered enzymatic biofuel cell based on amylases and glucose oxidase multi-immobilized bioanode
New Biotechnol.
30
531-535
2013
Albifimbria verrucaria (Q12737)
Kataoka, K.; Ito, T.; Okuda, Y.; Sakai, Y.; Yamashita, S.; Sakurai, T.
Roles of the indole ring of Trp396 covalently bound with the imidazole ring of His398 coordinated to type I copper in bilirubin oxidase
Biochem. Biophys. Res. Commun.
521
620-624
2020
Albifimbria verrucaria
Tsujimura, S.
From fundamentals to applications of bioelectrocatalysis bioelectrocatalytic reactions of FAD-dependent glucose dehydrogenase and bilirubin oxidase
Biosci. Biotechnol. Biochem.
83
39-48
2019
Albifimbria verrucaria
Tang, J.; Yan, X.; Huang, W.; Engelbrekt, C.; Duus, J.O.; Ulstrup, J.; Xiao, X.; Zhang, J.
Bilirubin oxidase oriented on novel type three-dimensional biocathodes with reduced graphene aggregation for biocathode
Biosens. Bioelectron.
167
112500
2020
Albifimbria verrucaria
Takimoto, D.; Tsujimura, S.
Oxygen reduction reaction activity and stability of electrochemically deposited bilirubin oxidase
Chem. Lett.
47
1269-1271
2018
Albifimbria verrucaria
-
Akter, M.; Tokiwa, T.; Shoji, M.; Nishikawa, K.; Shigeta, Y.; Sakurai, T.; Higuchi, Y.; Kataoka, K.; Shibata, N.
Redox potential-dependent formation of an unusual His-Trp bond in bilirubin oxidase
Chemistry
24
18052-18058
2018
Albifimbria verrucaria
Rawal, R.; Kharangarh, P.R.; Dawra, S.; Bhardwaj, P.
Synthesis, characterization and immobilization of bilirubin oxidase nanoparticles (BOxNPs) with enhanced activity Application for serum bilirubin determination in jaundice patients
Enzyme Microb. Technol.
143
109716
2021
Albifimbria verrucaria
Gross, A.J.; Chen, X.; Giroud, F.; Travelet, C.; Borsali, R.; Cosnier, S.
Redox-active glyconanoparticles as electron shuttles for mediated electron transfer with bilirubin oxidase in solution
J. Am. Chem. Soc.
139
16076-16079
2017
Albifimbria verrucaria
Bayineni, V.K.; Suresh, S.; Sharma, A.; Kadeppagari, R.K.
Improvement of bilirubin oxidase productivity of Myrothecium verrucaria and studies on the enzyme overproduced by the mutant strain in the solid-state fermentation
J. Gen. Appl. Microbiol.
64
68-75
2018
Albifimbria verrucaria, Albifimbria verrucaria MTCC 2140
Tokiwa, T.; Shoji, M.; Sladek, V.; Shibata, N.; Higuchi, Y.; Kataoka, K.; Sakurai, T.; Shigeta, Y.; Misaizu, F.
Structural changes of the trinuclear copper center in bilirubin oxidase upon reduction
Molecules
24
76
2018
Albifimbria verrucaria
Hasan, M.Q.; Kuis, R.; Narayanan, J.S.; Slaughter, G.
Fabrication of highly effective hybrid biofuel cell based on integral colloidal platinum and bilirubin oxidase on gold support
Sci. Rep.
8
16351
2018
Albifimbria verrucaria
Koval, T.; Svecova, L.; Ostergaard, L.H.; Skalova, T.; Duskova, J.; Hasek, J.; Kolenko, P.; Fejfarova, K.; Stransky, J.; Trundova, M.; Dohnalek, J.
Trp-His covalent adduct in bilirubin oxidase is crucial for effective bilirubin binding but has a minor role in electron transfer
Sci. Rep.
9
13700
2019
Albifimbria verrucaria (Q12737), Albifimbria verrucaria, Albifimbria verrucaria ATCC24571 (Q12737)
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