Information on EC 1.1.3.9 - galactose oxidase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
1.1.3.9
-
RECOMMENDED NAME
GeneOntology No.
galactose oxidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
D-galactose + O2 = D-galacto-hexodialdose + H2O2
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Galactose metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
D-galactose:oxygen 6-oxidoreductase
A copper protein.
CAS REGISTRY NUMBER
COMMENTARY hide
9028-79-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
strain NRRL 2903, expression in Pichia pastoris
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Polyporus circinatus
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,5-anhydrogalactitol + O2
? + H2O2
show the reaction diagram
Polyporus circinatus
-
-
-
?
1-methyl-alpha-D-galactopyranoside + O2
1-O-methyl-alpha-D-galacto-hexodialdose + H2O2
show the reaction diagram
1-O-methyl-alpha-D-galactopyranoside + O2
1-O-methyl-alpha-D-galacto-hexodialdose + H2O2
show the reaction diagram
-
-
-
-
?
1-O-methyl-alpha-D-galactosylpyranoside + O2
1-O-methyl-alpha-D-galacto-hexodialdose + H2O2
show the reaction diagram
1-O-methyl-alpha-D-glucosylpyranoside + O2
1-O-methyl-alpha-D-gluco-hexodialdose + H2O2
show the reaction diagram
1-O-methyl-beta-D-galactosylpyranoside + O2
1-O-methyl-beta-D-galacto-hexodialdose + H2O2
show the reaction diagram
1-O-methyl-beta-D-glucosylpyranoside + O2
1-O-methyl-beta-D-gluco-hexodialdose + H2O2
show the reaction diagram
1-O-methyl-D-galactopyranoside + O2
?
show the reaction diagram
-
112% of the activity with D-galactose
-
-
?
2 raffinose + 2 O2
6''-aldehydoraffinose + 6''-carboxyraffinose + H2O2 + H2O
show the reaction diagram
2-deoxy-D-galactose + O2
2-deoxy-D-galacto-hexodialdose + H2O2
show the reaction diagram
2-glycerol-alpha-D-galactosylpyranoside + O2
2-glycerol-alpha-D-galactosyl-hexodialdose + H2O2
show the reaction diagram
Polyporus circinatus
-
low activity
-
?
2-methylene-1,3-propanediol + O2
?
show the reaction diagram
-
-
-
?
3-bromo-1,2-propanediol + O2
? + H2O2
show the reaction diagram
-
-
S isomer
?
3-bromobenzyl alcohol + O2
3-bromobenzaldehyde + H2O2
show the reaction diagram
-
-
-
?
3-chloro-1,2-propanediol + O2
? + H2O2
show the reaction diagram
-
only R isomer will have the correct orientation to react with the enzyme
S isomer
?
3-chlorobenzyl alcohol + O2
3-chlorobenzaldehyde + H2O2
show the reaction diagram
-
-
-
?
3-fluoro-1-phenylethanol + O2
?
show the reaction diagram
-
-
-
-
?
3-fluorobenzyl alcohol + O2
3-fluorobenzaldehyde + H2O2
show the reaction diagram
-
-
-
?
3-methoxybenzyl alcohol + O2
3-methoxybenzaldehyde + H2O2
show the reaction diagram
-
-
-
?
3-methoxybenzyl alcohol + O2
? + H2O2
show the reaction diagram
-
-
-
-
?
3-nitrobenzyl alcohol + O2
3-nitrobenzaldehyde + H2O2
show the reaction diagram
-
-
-
?
4-(methylthio)benzyl alcohol + O2
4-(methylthio)benzaldehyde + H2O2
show the reaction diagram
-
-
-
?
4-(trifluoromethyl)benzyl alcohol + O2
4-(trifluoromethyl)benzaldehyde + H2O2
show the reaction diagram
-
-
-
?
4-bromobenzyl alcohol + O2
4-bromobenzaldehyde + H2O2
show the reaction diagram
-
-
-
?
4-chlorobenzyl alcohol + O2
4-chlorobenzaldehyde + H2O2
show the reaction diagram
-
-
-
?
4-fluorobenzyl alcohol + O2
4-fluorobenzaldehyde + H2O2
show the reaction diagram
-
-
-
?
4-iodobenzyl alcohol + O2
4-iodobenzaldehyde + H2O2
show the reaction diagram
-
-
-
?
4-methoxybenzyl alcohol + O2
4-methoxybenzaldehyde + H2O2
show the reaction diagram
-
-
-
?
4-methoxybenzyl alcohol + O2
? + H2O2
show the reaction diagram
-
-
-
-
?
4-methylbenzyl alcohol + O2
4-methylbenzaldehyde + H2O2
show the reaction diagram
-
-
-
?
4-nitrobenzyl alcohol + O2
4-nitrobenzaldehyde + H2O2
show the reaction diagram
-
-
-
?
4-nitrobenzyl alcohol + O2
? + H2O2
show the reaction diagram
-
-
-
-
?
4-O-beta-D-glucopyranosyl-D-glucose + O2
4-O-beta-D-glucopyranosyl-D-gluco-hexodialdose + H2O2
show the reaction diagram
acetol + O2
pyruvaldehyde + H2O2
show the reaction diagram
-
-
-
?
allyl alcohol + O2
acrolein + H2O2
show the reaction diagram
-
only extracelluar enzyme, low activity
-
?
alpha-D-talose + O2
alpha-D-talo-hexodialdose + H2O2
show the reaction diagram
benzene-1,2-diamine + O2
? + H2O2
show the reaction diagram
-
very low activity
-
?
benzyl alcohol + O2
? + H2O2
show the reaction diagram
-
-
-
-
?
benzyl alcohol + O2
benzaldehyde + H2O2
show the reaction diagram
benzylalcohol + O2
benzaldehyde + H2O2
show the reaction diagram
-
substrate reaction profiling
-
-
?
beta-D-galactopyranosyl-(1-6)-beta-D-galactopyranosyl-(1-4)-D-glucose + O2
? + H2O2
show the reaction diagram
Polyporus circinatus
-
-
-
?
beta-D-galactosyl-(1-6)-beta-D-galactopyranoside + O2
? + H2O2
show the reaction diagram
Polyporus circinatus
-
-
-
?
beta-D-lactose + O2
beta-D-lacto-hexodialdose + H2O2
show the reaction diagram
beta-hydroxypyruvate + O2
2,3-dioxopropionate + H2O2
show the reaction diagram
-
only extracellular enzyme, low activity
-
?
beta-thiodigalactoside + O2
? + H2O2
show the reaction diagram
Polyporus circinatus
-
-
-
?
beta-thiogalactoside + O2
beta-thiogalacto-hexodialdose + H2O2
show the reaction diagram
Polyporus circinatus
-
more rapidly oxidized than beta-D-galactose
-
?
ceramide dihexoside + O2
? + H2O2
show the reaction diagram
-
higher activity than free substrate, very low activity as vesicle-bound substrate
-
?
ceramide trihexoside + O2
? + H2O2
show the reaction diagram
-
vesicle-bound and free substrate
-
?
D-Gal-beta-(1-3)-D-Gal-beta-(1-1)-L-Gro + O2
? + H2O2
show the reaction diagram
-
low activity
-
?
D-Gal-beta-(1-3)-D-Gal-beta-(1-3)-D-Gal-(1-1)-L-Gro + O2
? + H2O2
show the reaction diagram
-
low activity
-
?
D-Gal-beta-(1-3)-D-Gal-beta-(1-6)-D-Gal-beta-(1-1)-L-Gro + O2
? + H2O2
show the reaction diagram
-
lower activity than with a reversed beta-1-6-linkage
-
?
D-Gal-beta-(1-3)-[D-Gal-beta-(1-6)]-D-Gal-beta-(1-1)-L-Gro + O2
? + H2O2
show the reaction diagram
-
best oligosaccharide oxidized
-
?
D-Gal-beta-(1-6)-D-Gal-beta-(1-1)-L-Gro + O2
? + H2O2
show the reaction diagram
-
faster oxidation than corresponding beta-1-3-linked components
-
?
D-Gal-beta-(1-6)-D-Gal-beta-(1-3)-D-Gal-beta-(1-1)-L-Gro + O2
? + H2O2
show the reaction diagram
-
improved activity
-
?
D-galactopyranose + ferricyanide
D-galacto-hexodialdose + ferrocyanide
show the reaction diagram
-
ferricyanide poorly replaces O2 as electron acceptor
-
?
D-galactosamine + O2
? + H2O2
show the reaction diagram
D-galactose + O2
D-galacto-hexodialdose + H2O2
show the reaction diagram
D-galactosylpyranoside + O2
D-galacto-hexodialdose + H2O2
show the reaction diagram
D-glucosylpyranoside + O2
D-gluco-hexodialdose + H2O2
show the reaction diagram
-
very low activity
-
?
D-xylose + O2
? + H2O2
show the reaction diagram
-
very low activity
-
?
dihydroxyacetone + O2
3-hydroxy-2-oxo-propionaldehyde + H2O2
show the reaction diagram
dihydroxyacetone + O2
?
show the reaction diagram
-
-
-
-
?
fetuin + O2
? + H2O2
show the reaction diagram
-
bovine fetuin, native or desialylated
-
?
Forssman glycolipid + O2
? + H2O2
show the reaction diagram
-
higher activity as vesicle-bound substrate, very low activity as free substrate
-
?
Gal-beta-(1-3)-[Fuc-alpha-(1-2)]-GalNAcol + O2
? + H2O2
show the reaction diagram
-
no oxidation of oligosaccharides containing N-acetylgalactosamine at the non-reducing end
-
?
galactan + O2
? + H2O2
show the reaction diagram
-
derived from snail, Lymnea stagnalis galactan best substrate
-
?
galactogen + O2
? + H2O2
show the reaction diagram
-
substrate from Helix pomatia, galactose oxidase acts upon a specific subterminal nonreducing D-galactosyl residue
-
?
galactolipid + O2
? + H2O2
show the reaction diagram
-
-
-
?
galactose 1-phosphate + O2
? + H2O2
show the reaction diagram
Polyporus circinatus
-
very low activity
-
?
galactose-4-SO3Na + O2
? + H2O2
show the reaction diagram
-
very low activity
-
?
galactose-6-SO3Na + O2
? + H2O2
show the reaction diagram
-
very low activity
-
?
ganglioside + O2
? + H2O2
show the reaction diagram
globoside + O2
? + H2O2
show the reaction diagram
-
human and porcine globoside, vesicle-bound and free substrate, best substrate tested
-
?
glyceraldehyde + O2
? + H2O2
show the reaction diagram
-
70% of the activity with glycolaldehyde
-
-
?
glycerol + O2
(S)-glyceraldehyde + H2O2
show the reaction diagram
glycoaldehyde + O2
glyoxal + H2O2
show the reaction diagram
-
only extracellular enzyme, low activity
-
?
glycolaldehyde + O2
glyoxal + H2O2
show the reaction diagram
glycoprotein + O2
? + H2O2
show the reaction diagram
-
-
-
?
guar + O2
? + H2O2
show the reaction diagram
guar gum + O2
? + H2O2
show the reaction diagram
-
significant activity
-
?
guaran + O2
? + H2O2
show the reaction diagram
hexadecyl-(ethyleneglycol)13-D-galactose + O2
hexadecyl-(ethyleneglycol)13-D-galacto-hexodialdose + H2O2
show the reaction diagram
-
-
-
-
?
hexadecyl-(ethyleneglycol)20-D-galactose + O2
hexadecyl-(ethyleneglycol)20-D-galacto-hexodialdose + H2O2
show the reaction diagram
-
-
-
-
?
hexadecyl-(ethyleneglycol)6-D-galactose + O2
hexadecyl-(ethyleneglycol)6-D-galacto-hexodialdose + H2O2
show the reaction diagram
-
-
-
-
?
hexadecyl-(ethyleneglycol)9-D-galactose + O2
hexadecyl-(ethyleneglycol)9-D-galacto-hexodialdose + H2O2
show the reaction diagram
-
-
-
-
?
isopropyl-beta-D-thiogalactosylpyranoside + O2
isopropyl-beta-D-thiogalactosyl-hexodialdose + H2O2
show the reaction diagram
Polyporus circinatus
-
43% of the activity compared to D-galactose
-
?
L-glucose + O2
L-gluco-hexodialdose + H2O2
show the reaction diagram
-
very low activity
-
?
lactose + O2
?
show the reaction diagram
-
4% of the activity with D-galactose
-
-
?
lactose + O2
? + H2O2
show the reaction diagram
Polyporus circinatus
-
very low activity
-
?
maltose + O2
? + H2O2
show the reaction diagram
-
very low activity
-
?
melibiitol + O2
? + H2O2
show the reaction diagram
Polyporus circinatus
-
-
-
?
melibionic acid + O2
? + H2O2
show the reaction diagram
Polyporus circinatus
-
low activity
-
?
melibiose + O2
?
show the reaction diagram
-
103% of the activity with D-galactose
-
-
?
melibiose + O2
? + H2O2
show the reaction diagram
methyl alpha-D-galactopyranoside + O2
methyl alpha-D-galacto-hexodialdo-1,5-pyranoside + H2O2
show the reaction diagram
-
investigation of the optimal reaction conditions (reaction medium, temperature, concentration and combinations of galactose oxidase, catalase, and horseradish peroxidase are used as variables) to degrade methyl alpha-D-galactopyranoside to alpha-D-galacto-hexodialdo-1,5-pyranoside and thereby reduce byproduct formation. Optimal combination of the 3 enzymes gives methyl alpha-D-galacto-hexodialdo-1,5-pyranoside in approximately 90% yield
-
-
?
methyl alpha-D-galactopyranoside + O2
methyl alpha-D-galacto-hexodialdose + H2O2
show the reaction diagram
-
-
-
-
?
methyl beta-D-thiogalactosylpyranoside + O2
methyl beta-D-thiogalacto-hexodialdose + H2O2
show the reaction diagram
Polyporus circinatus
-
-
-
?
mucin + O2
? + H2O2
show the reaction diagram
-
bovine submaxillary mucin, native and desialylated
-
-
N-acetyl-D-galactosamine + O2
? + H2O2
show the reaction diagram
o-nitrophenyl beta-D-galactoside + O2
1-O-(o-nitrophenyl)-alpha-D-galactohexodialdose + H2O2
show the reaction diagram
p-nitrophenyl alpha-D-galactoside + O2
1-O-(p-nitrophenyl)-alpha-D-galactohexodialdose + H2O2
show the reaction diagram
-
more reactive than p-nitrophenyl-beta-D-galactoside
-
?
p-nitrophenyl beta-D-galactoside + O2
1-O-(p-nitrophenyl)-beta-D-galactohexodialdose + H2O2
show the reaction diagram
-
less reactive than p-nitrophenyl-alpha-D-galactoside
-
?
planteose + O2
? + H2O2
show the reaction diagram
Polyporus circinatus
-
-
-
?
raffinose + O2
?
show the reaction diagram
raffinose + O2
? + H2O2
show the reaction diagram
sphingoglycolipid + O2
? + H2O2
show the reaction diagram
-
-
-
?
stachyose + O2
? + H2O2
show the reaction diagram
sucrose + O2
? + H2O2
show the reaction diagram
-
very low activity
-
?
talose + O2
? + H2O2
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
2 raffinose + 2 O2
6''-aldehydoraffinose + 6''-carboxyraffinose + H2O2 + H2O
show the reaction diagram
2-deoxy-D-galactose + O2
2-deoxy-D-galacto-hexodialdose + H2O2
show the reaction diagram
4-O-beta-D-glucopyranosyl-D-glucose + O2
4-O-beta-D-glucopyranosyl-D-gluco-hexodialdose + H2O2
show the reaction diagram
alpha-D-talose + O2
alpha-D-talo-hexodialdose + H2O2
show the reaction diagram
beta-D-galactopyranosyl-(1-6)-beta-D-galactopyranosyl-(1-4)-D-glucose + O2
? + H2O2
show the reaction diagram
Polyporus circinatus
-
-
-
?
beta-D-galactosyl-(1-6)-beta-D-galactopyranoside + O2
? + H2O2
show the reaction diagram
Polyporus circinatus
-
-
-
?
beta-D-lactose + O2
beta-D-lacto-hexodialdose + H2O2
show the reaction diagram
ceramide dihexoside + O2
? + H2O2
show the reaction diagram
-
higher activity than free substrate, very low activity as vesicle-bound substrate
-
?
ceramide trihexoside + O2
? + H2O2
show the reaction diagram
-
vesicle-bound and free substrate
-
?
D-galactosamine + O2
? + H2O2
show the reaction diagram
D-galactose + O2
D-galacto-hexodialdose + H2O2
show the reaction diagram
D-galactosylpyranoside + O2
D-galacto-hexodialdose + H2O2
show the reaction diagram
D-glucosylpyranoside + O2
D-gluco-hexodialdose + H2O2
show the reaction diagram
-
very low activity
-
?
D-xylose + O2
? + H2O2
show the reaction diagram
-
very low activity
-
?
dihydroxyacetone + O2
3-hydroxy-2-oxo-propionaldehyde + H2O2
show the reaction diagram
fetuin + O2
? + H2O2
show the reaction diagram
-
bovine fetuin, native or desialylated
-
?
Forssman glycolipid + O2
? + H2O2
show the reaction diagram
-
higher activity as vesicle-bound substrate, very low activity as free substrate
-
?
Gal-beta-(1-3)-[Fuc-alpha-(1-2)]-GalNAcol + O2
? + H2O2
show the reaction diagram
-
no oxidation of oligosaccharides containing N-acetylgalactosamine at the non-reducing end
-
?
galactolipid + O2
? + H2O2
show the reaction diagram
-
-
-
?
ganglioside + O2
? + H2O2
show the reaction diagram
globoside + O2
? + H2O2
show the reaction diagram
-
human and porcine globoside, vesicle-bound and free substrate, best substrate tested
-
?
glycoprotein + O2
? + H2O2
show the reaction diagram
-
-
-
?
guar gum + O2
? + H2O2
show the reaction diagram
-
significant activity
-
?
guaran + O2
? + H2O2
show the reaction diagram
maltose + O2
? + H2O2
show the reaction diagram
-
very low activity
-
?
melibiose + O2
? + H2O2
show the reaction diagram
mucin + O2
? + H2O2
show the reaction diagram
-
bovine submaxillary mucin, native and desialylated
-
-
N-acetyl-D-galactosamine + O2
? + H2O2
show the reaction diagram
planteose + O2
? + H2O2
show the reaction diagram
Polyporus circinatus
-
-
-
?
raffinose + O2
? + H2O2
show the reaction diagram
sphingoglycolipid + O2
? + H2O2
show the reaction diagram
-
-
-
?
stachyose + O2
? + H2O2
show the reaction diagram
sucrose + O2
? + H2O2
show the reaction diagram
-
very low activity
-
?
additional information
?
-
-
the oxidized form of the enzyme catalyzes the two-electron oxidation of a broad range of primary alcohols to corresponding aldehydes with the concomitant reduction of O2 to H2O2
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyrroloquinoline quinone
-
one pyrroquinoline covalently bound to one molecule, additional two-electron redox center
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
copper
-
copper content: 1.2 mol of Cu per mol of wild-type enzyme, 0.96 mol of Cu per mol of mutant enzyme W290F, 1 mol Cu per mol of mutant enzyme W290G, 1.3 mol of Cu per mol of mutant enzyme W290H
Cu3+
-
first evidence of oxidation by the Cu(III) enzyme with concomitant formation of a Cu(I) enzyme
Fe
-
1 atom iron per mol enzyme
additional information
-
the monomeric enzyme containing a mononuclear copper ionis coordinated with an unusually stable cysteinyl-tyrosine protein free radical, removal of copper and free radicals inactivates the enzyme
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2'-dipyridyl
-
1 mM, 67% inhibition
2,4,6-Trinitrobenzenesulfonic acid
-
53% inhibition at 1 mM
2-deoxy-D-galactose
Polyporus circinatus
-
competitive inhibition
2-mercaptoethanol
8-hydroxyquinoline
-
1 mM, 50% inhibition
AgNO3
-
complete inhibition at 0.1 mM
Co2+
-
1 mM, 50% inhibition
D-Galactono-1,4-lactone
-
23.7% inhibition at 0.25 mM
D-galactosamine
Polyporus circinatus
-
competitive inhibition
D-galacturonic acid
-
66.8% inhibition at 0.25 mM
diethyldithiocarbamate
dithiothreitol
-
40% inhibition at 1 mM
Fe2+
-
1 mM, 17% inhibition
ferricyanide
-
inhibition at 0.001 mM, activation at 1 mM
ganglioside
Polyporus circinatus
-
bovine brain and Tay-Sachs disease gangliosides: competitive inhibition
GSH
-
complete inhibition at 10 mM
hydrazine
-
1 mM, 41% inhibition
hydroxylamine
iodoacetamide
iodoacetic acid
-
slight inactivation
L-ascorbic acid
-
58% inhibition at 0.1 mM
Mg2+
-
1 mM, 50% inhibition
Monoiodoacetic acid
-
1 mM, 40% inhibition
N-acetylgalactosamine
Polyporus circinatus
-
competitive inhibition
N-bromosuccinimide
NH2OH
-
complete inhibition at 0.1 mM
Ni2+
-
1 mM, 50% inhibition
o-phenanthroline
p-chloromercuribenzoate
-
22% inhibition at 0.1 mM
phenylhydrazine
-
1 mM, 91% inhibition
Semicarbazide
-
1 mM, 44% inhibition
Sodium dithionite
-
73% inhibition at 0.1 mM
Superoxide dismutase
-
50% inhibition at 50 pM
-
trans-(2)-phenylcyclopropylcarbinol
-
mechanism-based inactivator
Zn2+
-
1 mM, 33% inhibition
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
chloroiridate
-
marked increase of activity
Coprimus cinereus peroxidase
-
13fold activation
-
EDTA
-
up to 7fold activation
ferricyanide
superoxide
-
stimulation of O2 uptake
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
16 - 56
1-methyl-alpha-D-galactopyranoside
77 - 440
1-O-methyl-alpha-D-galactopyranoside
29 - 57
2-methylene-1,3-propanediol
0.0021 - 2950
D-galactose
7.5
glyceraldehyde
-
-
13.2
glycolaldehyde
-
-
additional information
additional information
-
steady-state kinetics, isotope kinetics, kinetic analysis
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
457 - 1548
1-methyl-alpha-D-galactopyranoside
325
1-O-methyl beta-D-galactopyranoside
Hypomyces rosellus
-
Cu(II)-bound H2O is required in the protonated aquo state for catalysis and is responsible for the kinetic solvent isotope effect
20 - 1165
1-O-methyl-alpha-D-galactopyranoside
1440
1-O-methyl-beta-D-galactopyranoside
Hypomyces rosellus
-
-
166 - 283
2-methylene-1,3-propanediol
0.011 - 3400
D-galactose
13.7 - 58
guar
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00019 - 32
D-galactose
71
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.3
2-deoxy-D-galactose
Polyporus circinatus
-
-
0.073
D-galactosamine
Polyporus circinatus
-
-
0.002 - 0.12
ganglioside
5.7
iodoacetamide
-
pseudo-first-order kinetics, pH-independent
0.047
N-acetylgalactosamine
Polyporus circinatus
-
-
5.6
trans-(2)-phenylcyclopropylcarbinol
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.035
-
mutant C228G
0.54
-
mutant W290H
116
-
intracellular enzyme
1093
-
recombinant wild-type enzyme
1120
-
native wild-type enzyme
additional information
-
enzyme activity in filtrates of the different isolates, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
-
recombinant enzyme, optimal condition for glyoxal production
6
-
assay at
6.5 - 7.5
-
maximum kcat/Km
6.5
-
oxidation of glycolaldehyde
7 - 7.5
-
dependent on the strain
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10.5
-
tested range, decrease of kcat/Km by 2 orders of magnitude compared to maximum kcat/Km
6 - 9
-
pH 6.0: about 40% of maximal activity, pH 9.0: about 65% of maximal activity
6 - 7
-
pH 6: about 80% of maximal activity, pH 7: about 25% loss of activity, oxidation of glycolaldehyde
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 50
-
20C: about 90% of maximal activity, 50C: about 60% of maximal activity
30 - 60
-
50% of maximal activity at 60C
40 - 60
-
40C: about 65% of maximal activity, 60C: about 80% of maximal activity, oxidation of glycolaldehyde
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.2
-
isoelectric focusing, pH-range: 3.5-10
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
maximum galactose oxidase production (approximately 4.0 U/ml) is obtained when fermentation is carried out at 25C, with orbital shaking (100 rpm) and an initial medium of pH 7.0, for 96 h, using a 2% (v/v) inoculum made from a homogenized four-day-old liquid culture, in the presence of copper, manganese, and magnesium
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24000
-
SDS-PAGE
42100
Polyporus circinatus
-
high speed membrane osmometer
42400
Polyporus circinatus
-
sedimentation equilibrium centrifugation
49000
-
gel filtration
65000
-
SDS-PAGE
68500
-
recombinant galactose oxidase without the pro sequence, SDS-PAGE
72000
-
gel filtration, intracellular form
90000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
2 * 24000, SDS-PAGE
monomer
additional information
-
the active site structure consists of two one-electron redox units involving residues Y495, H496, H581, Y272, and W290, a Cu2+ ion, and a crosslinked Y272-C228 radical cofactor, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
-
cleavage of a secretion signal sequence, copper-dependent cleavage of an N-terminal pro sequence
side-chain modification
-
copper-dependent formation of a C228-Y272 thioether bond, generation of the Y272 radical
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystals of W290F and W290G are grown from protein as isolated, using hanging drop vapor diffusion from 1 to 2 M (NH4)2SO4 and 0.1 M sodium acetate at pH 4.0-5.0. To prepare the azide complex, wild-type galactose oxidase is dialyzed into 50 mM PIPES (pH 7.0) for 2 h, before addition of 20 mM sodium iethyl dithiocarbamate (DDC). The copper chelator DDC promotes the formation of orthorhombic crystals, which have more favorable crystal packing for substrate soaking experiments. The copper-free protein is crystallized in 13.5% PEG 8000, 200 mM calcium acetate, and 100 mM MES (pH 6.3)
-
sitting drop method. 2-5 mg/ml of enzyme mixed with equal volume of well solution. Crystals are soaked in oxygen-free Cu2+ solutions
-
mutant C383S
-
ammonium sulfate precipitation, 1.7 A resolution
-
hanging drop vapor diffusion, 1.7 A resolution
-
sitting drop vapour diffusion, native, recombinant and mutant enzymes
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
40C, 60 min, stable
687852
6
-
40C, 60 min, about 5% loss of activity
687852
7 - 7.5
-
most stable at
672854
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 60
-
30 min, stable
40
-
pH 6.5, 60 min, about 10% loss of activity
50
-
pH 6.5, 60 min, about 20% loss of activity
65
-
10 min, about 40% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
resistant to chemical denaturation, low thermal stability
-
sensitive to inactivation by guanidium-HCl, recovery of activity upon dilution
-
stable at room temperature, sensitive to H2O2
Polyporus circinatus
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-17C, stable for 6 months
-
-70C, 1 year stable
-
4C, 0.01 M phosphate buffer, pH 7.0, 0.1 mg/ml protein, 30% decrease of activity in 1 year
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
covalently immobilized to chemically modified porous silica particles
-
Ni-charged resin column chromatography
-
purification of recombinant galactose oxidase under strict copper-free conditions
-
purification using immobilized biomimetic ligands or D(+)-galactosamine, two-step purification procedure using immobilized 1-amino-1-deoxy-beta-D-galactose
-
recombinant enzyme
-
recombinant enzyme; to homogeneity, 1step chromatography
-
the enzyme from commercial preparation is further purified by gel filtration
-
to homogeneity, 1step chromoatography
Polyporus circinatus
-
to homogeneity, affinity chromatography
-
to homogeneity, chromatography techniques
to homogeneity, intra- and extracellular forms, 1step chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
computational active-site models for galactose oxidase for the distinctive oxidation states (Cu2+ protein complex/radical), additional models including solvent molecules and second coordination sphere residues
-
expressed in Escherichia coli BL21 cells
-
expression in Aspergillus nidulans
-
expression in Aspergillus oryzae, Fusarium venenatum
-
expression in Pichia pastoris
-
functional expression of wild-type and mutant enzymes in Escherichia coli strain BL21 Star (DE3) cytoplasm, best in strain BL21 Star (DE3) at 25C, induction for 4-6 h is optimal
-
high level expression in Pichia pastoris
-
recombinant expression of galactose oxidase in Pichia pastoris
-
recombinant expression of galactose oxidase without the pro sequence. Reaction with excess Cu2+ leads to formation of the C228-Y272 thioether bond both under aerobic and anaerobic conditions, the formation of the Y272 radical occures solely under aerobic conditions (posttranslational modifications)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C383A
-
site-directed mutagenesis, the mutant shows 39% of wild-typ enzyme activity
C383D
-
site-directed mutagenesis, the mutant shows 26% of wild-typ enzyme activity
C383E
-
site-directed mutagenesis, the mutant shows 130% of wild-typ enzyme activity
C383F
-
site-directed mutagenesis, the mutant shows 8% of wild-typ enzyme activity
C383G
-
site-directed mutagenesis, the mutant shows 70% of wild-typ enzyme activity
C383H
-
site-directed mutagenesis, the mutant shows 35% of wild-typ enzyme activity
C383I
-
site-directed mutagenesis, the mutant shows 3% of wild-typ enzyme activity
C383K
-
site-directed mutagenesis, the mutant shows 115% of wild-typ enzyme activity
C383L
-
site-directed mutagenesis, the mutant shows 41% of wild-typ enzyme activity
C383M
-
site-directed mutagenesis, the mutant shows 75% of wild-typ enzyme activity
C383N
-
site-directed mutagenesis, the mutant shows 6% of wild-typ enzyme activity
C383P
-
site-directed mutagenesis, the mutant shows 18% of wild-typ enzyme activity
C383Q
-
site-directed mutagenesis, the mutant shows 13% of wild-typ enzyme activity
C383R
-
site-directed mutagenesis, the mutant shows 0.2% of wild-typ enzyme activity
C383S
-
site-directed mutagenesis, the mutant shows 160% of wild-typ enzyme activity
C383T
-
site-directed mutagenesis, the mutant shows 32% of wild-typ enzyme activity
C383V
-
site-directed mutagenesis, the mutant shows 4% of wild-typ enzyme activity
C383W
-
site-directed mutagenesis, the mutant shows 0.001% of wild-typ enzyme activity
C383Y
-
site-directed mutagenesis, the mutant cannt be isolated
G195E
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme
M70V
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme
N535D
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme
V494A
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme
W290F
-
kcat/KM for D-galactose is 49fold lower than wild-type value, kcat/Km for 2-methylene-1,3-propanediol is 1.2fold higher than wild-type value
W290G
-
kcat/KM for D-galactose is 6370fold lower than wild-type value
W290H
-
kcat/KM for D-galactose is 1180fold lower than wild-type value
C383/Y436H
-
decrease in Km-value, increase in kcat-value
C383A
-
slight decrease in Km-value, decrease in kcat-value
C383H/V494A
-
decrease in Km-value, decrease in kcat-value
C383S
-
decrease in Km-value
C383S/Y436A
-
decrease in Km-value, decrease in kcat-value of 1-methyl-alpha-D-galactose, increase in kcat-value of D-galactose
C383S/Y436H/V494A
-
decrease in Km-value, increase in kcat-value
P463I
-
random mutagenesis, the mutant shows altered substrate specificity with several substrates compared to the wild-type enzyme
P463V
-
random mutagenesis, the mutant shows altered substrate specificity with several substrates compared to the wild-type enzyme
R330K
-
the mutant shows increased activity with D-fructose compared to the wild-type enzyme
R330K/W290F/Q406E/Y405F
-
the mutant shows 136fold increased activity with D-fructose, and increased activity with mannose and N-acetylglucosamine compared to the wild-type enzyme
S10P/M70 V/P136/G195E/V494A/N535D
-
random mutagenesis, the enzyme mutant shows improved levels of recombinant expression of a more active and stable enzyme in Escherichia coli without any change in substrate range compared to the wild-type enzyme
V494A
-
slight decrease in Km-value, increase in kcat-value
W290F/R330K/Q406T
-
random mutagenesis, the mutant shows improved activity toward Glc compared to the wild-type enzyme
Y405F/Q406E
-
random mutagenesis, the mutant shows altered substrate specificity with several substrates compared to the wild-type enzyme
Y405F/Q406Y
-
random mutagenesis, the mutant shows altered substrate specificity with several substrates compared to the wild-type enzyme
Y436A
-
Km- and kcat-values similar to wild type
Y436A/V494A
-
decrease in Km-value, increase in kcat-value
Y436H
-
slight decrease in Km-value, decrease in kcat-value
Y436H/V494A
-
slight decrease in kcat-value of 1-methyl-alpha-D-galactose, slight increase in kcat-value of D-galactose
C383A
-
slight decrease in Km-value, decrease in kcat-value
-
C383S
-
decrease in Km-value
-
V494A
-
slight decrease in Km-value, increase in kcat-value
-
Y436A
-
Km- and kcat-values similar to wild type
-
Y436H
-
slight decrease in Km-value, decrease in kcat-value
-
W290H
-
analysis from X-ray structure
Y272F
-
not capable of binding copper
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
biotechnology
-
technology: creating hybrid-system by immobilization of GOase on gold electrode, technology enables creation of biosensors and biofuel cells and studying electrochemically the catalytic mechanism of reactions for which free radicals and electron-transfer reactions are involved
diagnostics
-
determination of galactose or lactose concentration in complex biological fluids by immobilized galactose oxidase
molecular biology
-
glycoprotein labeling using engineered variants of galactose oxidase, overview
synthesis