BRENDA - Enzyme Database
show all sequences of 1.14.11.48

Purification and characterization of the FeII- and alpha-ketoglutarate-dependent xanthine hydroxylase from Aspergillus nidulans

Montero-Moran, G.M.; Li, M.; Rendon-Huerta, E.; Jourdan, F.; Lowe, D.J.; Stumpff-Kane, A.W.; Feig, M.; Scazzocchio, C.; Hausinger, R.P.; Biochemistry 46, 5293-5304 (2007)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
overproduction of His6-tagged enzyme in Aspergillus nidulans and Escherichia coli
Aspergillus nidulans
Inhibitors
Inhibitors
Commentary
Organism
Structure
6,8-dihydroxypurine
0.08 mM, 24% decrease in activity
Aspergillus nidulans
Co2+
0.04 mM, about 65% inhibition
Aspergillus nidulans
Cu2+
0.04 mM, complete inhibition
Aspergillus nidulans
Fe2+
depending on the experimental conditions, Fe2+ concentrations of above 0.025 mM can lead to enzyme inhibition of the recombinant enzyme isolated from Escherichia coli, whereas the enzyme from Aspergillus nidulans is not inhibited at Fe2+ concentrations up to 0.080 mM
Aspergillus nidulans
Mn2+
0.04 mM, about 80% inhibition
Aspergillus nidulans
N-oxalylglycine
competes with 2-oxoglutarate
Aspergillus nidulans
NaCl
0.5 M NaCl increases the Km of 2-oxoglutarate, increases the Km of xanthine and decreased the kcat
Aspergillus nidulans
Zn2+
0.04 mM, complete inhibition
Aspergillus nidulans
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0311
-
2-oxoglutarate
25°C, pH 7.4, recombinant enzyme isolated from Escherichia coli
Aspergillus nidulans
0.0452
-
xanthine
25°C, pH 7.4, recombinant enzyme isolated from Escherichia coli
Aspergillus nidulans
0.046
-
xanthine
30°C, pH 7.4, enzyme isolated from Aspergillus nidulans
Aspergillus nidulans
0.05
-
2-oxoglutarate
30°C, pH 7.4, enzyme isolated from Aspergillus nidulans
Aspergillus nidulans
0.16
-
2-oxoadipate
25°C, pH 7.4, recombinant enzyme isolated from Escherichia coli
Aspergillus nidulans
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Fe2+
reqired. Half-maximal activity at 0.007 mM when the recombinant apoprotein isolated from the Escherichia coli is used
Aspergillus nidulans
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
40000
-
12 * 40000, SDS-PAGE
Aspergillus nidulans
500000
-
dodecamer, gel filtration
Aspergillus nidulans
Organism
Organism
UniProt
Commentary
Textmining
Aspergillus nidulans
-
-
-
Posttranslational Modification
Posttranslational Modification
Commentary
Organism
glycoprotein
presence of O-glycosylation as well as N-glycosylation
Aspergillus nidulans
Purification (Commentary)
Purification (Commentary)
Organism
purified from both the fungal mycelium and recombinant Escherichia coli cells
Aspergillus nidulans
Source Tissue
Source Tissue
Commentary
Organism
Textmining
mycelium
-
Aspergillus nidulans
-
Storage Stability
Storage Stability
Organism
-80°C, stable for at least 2 months
Aspergillus nidulans
4°C, stored in 100 mM Tris buffer (pH 7) containing 300 mM NaCl, 250 mM imidazole, 15% glycerol, and 0.005 mM Fe2+, concentrated enzyme is stable for at least 5 months
Aspergillus nidulans
4°C, stored in 100 mM Tris buffer (pH 8.0) containing 300 mM NaCl, 250 mM imidazole, 1 mM EDTA, and 15% glycerol, stable for at least 1 month
Aspergillus nidulans
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
additional information
no activity with pyruvate, 2-oxobutyrate, phenyl pyruvate, and 4-hydroxyphenyl pyruvate
731289
Aspergillus nidulans
?
-
-
-
?
xanthine + 2-oxoadipate + O2
-
731289
Aspergillus nidulans
?
-
-
-
?
xanthine + 2-oxoglutarate + O2
the enzyme is highly specific for xanthine. On the basis of the spectroscopic assay using standard conditions with 12 nM enzyme, no activity is detected when the enzyme is assayed with 0.08, 0.1, or 0.2 mM hypoxanthine, 1-methylxanthine, 3-methylxanthine, 7-methylxanthine, 9-methylxanthine, purine, 6-methylpurine, 2-hydroxypurine, 8-hydroxypurine, 2,8-dihydroxypurine, 2-hydroxy-6-methylpurine, allopurinol, allantoin, or adenosine diphosphate
731289
Aspergillus nidulans
urate + succinate + CO2
-
-
-
?
Subunits
Subunits
Commentary
Organism
dodecamer
12 * 40000, SDS-PAGE
Aspergillus nidulans
Synonyms
Synonyms
Commentary
Organism
XanA
-
Aspergillus nidulans
xanthine/alpha-ketoglutarate dioxygenase
-
Aspergillus nidulans
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
7.6
-
2-oxoadipate
25°C, pH 7.4, recombinant enzyme isolated from Escherichia coli
Aspergillus nidulans
15
30
2-oxoglutarate
30°C, pH 7.4, value depending on preparation, enzyme isolated from Aspergillus nidulans
Aspergillus nidulans
15
30
xanthine
30°C, pH 7.4, value depending on preparation, enzyme isolated from Aspergillus nidulans
Aspergillus nidulans
66.5
-
2-oxoglutarate
25°C, pH 7.4, recombinant enzyme isolated from Escherichia coli
Aspergillus nidulans
71.4
-
xanthine
25°C, pH 7.4, recombinant enzyme isolated from Escherichia coli
Aspergillus nidulans
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
8
the recombinant enzyme isolated from Escherichia coli exhibits a narrow pH optimum of 7.0-8.0 with pH 7.4 yielding optimal activity for Tris, MOPS, MES, imidazole, and HEPES buffers
Aspergillus nidulans
7
-
enzyme isolated from Aspergillus nidulans
Aspergillus nidulans
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
7
11
stable
Aspergillus nidulans
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.00012
-
N-oxalylglycine
pH 7.4, 25°C
Aspergillus nidulans
Cloned(Commentary) (protein specific)
Commentary
Organism
overproduction of His6-tagged enzyme in Aspergillus nidulans and Escherichia coli
Aspergillus nidulans
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
6,8-dihydroxypurine
0.08 mM, 24% decrease in activity
Aspergillus nidulans
Co2+
0.04 mM, about 65% inhibition
Aspergillus nidulans
Cu2+
0.04 mM, complete inhibition
Aspergillus nidulans
Fe2+
depending on the experimental conditions, Fe2+ concentrations of above 0.025 mM can lead to enzyme inhibition of the recombinant enzyme isolated from Escherichia coli, whereas the enzyme from Aspergillus nidulans is not inhibited at Fe2+ concentrations up to 0.080 mM
Aspergillus nidulans
Mn2+
0.04 mM, about 80% inhibition
Aspergillus nidulans
N-oxalylglycine
competes with 2-oxoglutarate
Aspergillus nidulans
NaCl
0.5 M NaCl increases the Km of 2-oxoglutarate, increases the Km of xanthine and decreased the kcat
Aspergillus nidulans
Zn2+
0.04 mM, complete inhibition
Aspergillus nidulans
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.00012
-
N-oxalylglycine
pH 7.4, 25°C
Aspergillus nidulans
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0311
-
2-oxoglutarate
25°C, pH 7.4, recombinant enzyme isolated from Escherichia coli
Aspergillus nidulans
0.0452
-
xanthine
25°C, pH 7.4, recombinant enzyme isolated from Escherichia coli
Aspergillus nidulans
0.046
-
xanthine
30°C, pH 7.4, enzyme isolated from Aspergillus nidulans
Aspergillus nidulans
0.05
-
2-oxoglutarate
30°C, pH 7.4, enzyme isolated from Aspergillus nidulans
Aspergillus nidulans
0.16
-
2-oxoadipate
25°C, pH 7.4, recombinant enzyme isolated from Escherichia coli
Aspergillus nidulans
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
reqired. Half-maximal activity at 0.007 mM when the recombinant apoprotein isolated from the Escherichia coli is used
Aspergillus nidulans
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
40000
-
12 * 40000, SDS-PAGE
Aspergillus nidulans
500000
-
dodecamer, gel filtration
Aspergillus nidulans
Posttranslational Modification (protein specific)
Posttranslational Modification
Commentary
Organism
glycoprotein
presence of O-glycosylation as well as N-glycosylation
Aspergillus nidulans
Purification (Commentary) (protein specific)
Commentary
Organism
purified from both the fungal mycelium and recombinant Escherichia coli cells
Aspergillus nidulans
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
mycelium
-
Aspergillus nidulans
-
Storage Stability (protein specific)
Storage Stability
Organism
-80°C, stable for at least 2 months
Aspergillus nidulans
4°C, stored in 100 mM Tris buffer (pH 7) containing 300 mM NaCl, 250 mM imidazole, 15% glycerol, and 0.005 mM Fe2+, concentrated enzyme is stable for at least 5 months
Aspergillus nidulans
4°C, stored in 100 mM Tris buffer (pH 8.0) containing 300 mM NaCl, 250 mM imidazole, 1 mM EDTA, and 15% glycerol, stable for at least 1 month
Aspergillus nidulans
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
additional information
no activity with pyruvate, 2-oxobutyrate, phenyl pyruvate, and 4-hydroxyphenyl pyruvate
731289
Aspergillus nidulans
?
-
-
-
?
xanthine + 2-oxoadipate + O2
-
731289
Aspergillus nidulans
?
-
-
-
?
xanthine + 2-oxoglutarate + O2
the enzyme is highly specific for xanthine. On the basis of the spectroscopic assay using standard conditions with 12 nM enzyme, no activity is detected when the enzyme is assayed with 0.08, 0.1, or 0.2 mM hypoxanthine, 1-methylxanthine, 3-methylxanthine, 7-methylxanthine, 9-methylxanthine, purine, 6-methylpurine, 2-hydroxypurine, 8-hydroxypurine, 2,8-dihydroxypurine, 2-hydroxy-6-methylpurine, allopurinol, allantoin, or adenosine diphosphate
731289
Aspergillus nidulans
urate + succinate + CO2
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
dodecamer
12 * 40000, SDS-PAGE
Aspergillus nidulans
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
7.6
-
2-oxoadipate
25°C, pH 7.4, recombinant enzyme isolated from Escherichia coli
Aspergillus nidulans
15
30
2-oxoglutarate
30°C, pH 7.4, value depending on preparation, enzyme isolated from Aspergillus nidulans
Aspergillus nidulans
15
30
xanthine
30°C, pH 7.4, value depending on preparation, enzyme isolated from Aspergillus nidulans
Aspergillus nidulans
66.5
-
2-oxoglutarate
25°C, pH 7.4, recombinant enzyme isolated from Escherichia coli
Aspergillus nidulans
71.4
-
xanthine
25°C, pH 7.4, recombinant enzyme isolated from Escherichia coli
Aspergillus nidulans
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
8
the recombinant enzyme isolated from Escherichia coli exhibits a narrow pH optimum of 7.0-8.0 with pH 7.4 yielding optimal activity for Tris, MOPS, MES, imidazole, and HEPES buffers
Aspergillus nidulans
7
-
enzyme isolated from Aspergillus nidulans
Aspergillus nidulans
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
7
11
stable
Aspergillus nidulans
Other publictions for EC 1.14.11.48
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
731217
Li
Characterization of active sit ...
Aspergillus nidulans
Arch. Biochem. Biophys.
470
44-53
2008
-
-
1
-
10
-
1
24
-
1
-
-
-
1
-
-
1
-
-
-
-
-
2
-
-
1
-
-
24
1
-
-
-
-
-
-
-
-
1
-
-
10
-
-
1
-
24
-
1
-
-
-
-
-
1
-
-
-
-
2
-
1
-
-
24
1
-
-
-
-
-
-
-
24
24
731289
Montero-Moran
Purification and characterizat ...
Aspergillus nidulans
Biochemistry
46
5293-5304
2007
-
-
1
-
-
-
8
5
-
1
2
-
-
4
-
1
1
-
-
1
-
3
3
1
2
-
-
-
5
2
-
1
-
1
-
-
-
-
1
-
-
-
-
-
8
1
5
-
1
2
-
-
-
1
1
-
1
-
3
3
1
-
-
-
5
2
-
1
-
-
-
-
-
-
-
732527
Cultrone
Convergent evolution of hydrox ...
Aspergillus nidulans
Mol. Microbiol.
57
276-290
2005
-
-
1
-
-
-
1
2
-
1
-
1
-
1
-
-
-
-
-
-
-
-
1
-
2
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
2
-
1
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-