BRENDA - Enzyme Database show
show all sequences of 1.14.13.113

Biochemical characterization of the HpxO enzyme from Klebsiella pneumoniae, a novel FAD-dependent urate oxidase

O'Leary, S.E.; Hicks, K.A.; Ealick, S.E.; Begley, T.P.; Biochemistry 48, 3033-3035 (2009)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
cloning and heterologous overexpression as N-terminally six-His tagged protein
Klebsiella pneumoniae
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.042
-
Urate
pH 8.0, cosubstrate 1.5 mM NADH
Klebsiella pneumoniae
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
45000
-
recombinant enzyme including the affinity tag, determined by SDS-PAGE
Klebsiella pneumoniae
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
additional information
Klebsiella pneumoniae
urate oxidase can catalyze the slow conversion of NADPH to NADP+ in the absence of urate
?
-
-
-
urate + NADH + H+ + O2
Klebsiella pneumoniae
part of the purine catabolic pathway
5-hydroxyisourate + NAD+ + H2O
unstable in aqueous buffer, hydrolyzing spontaneously to 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Klebsiella pneumoniae
-
-
-
Purification (Commentary)
Commentary
Organism
enzyme is purified by Ni2+-nitrilotriacetic acid chromatography
Klebsiella pneumoniae
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
urate oxidase can catalyze the slow conversion of NADPH to NADP+ in the absence of urate
696321
Klebsiella pneumoniae
?
-
-
-
-
urate + NADH + H+ + O2
part of the purine catabolic pathway
696321
Klebsiella pneumoniae
5-hydroxyisourate + NAD+ + H2O
unstable in aqueous buffer, hydrolyzing spontaneously to 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline
-
-
?
urate + NADH + H+ + O2
urate hydroxylase shows selectivity (V/K ratio of 10) for NADH over NADPH
696321
Klebsiella pneumoniae
5-hydroxyisourate + NAD+ + H2O
unstable in aqueous buffer, hydrolyzing spontaneously to 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
42
-
Urate
pH 8.0, 1.5 mM NADH as cosubstrate
Klebsiella pneumoniae
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Klebsiella pneumoniae
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
the enzyme is a flavoprotein
Klebsiella pneumoniae
NADH
the enzyme shows selectivity (V/K ratio of 10) for NADH over NADPH
Klebsiella pneumoniae
NADPH
the enzyme shows selectivity (V/K ratio of 10) for NADH over NADPH
Klebsiella pneumoniae
Cloned(Commentary) (protein specific)
Commentary
Organism
cloning and heterologous overexpression as N-terminally six-His tagged protein
Klebsiella pneumoniae
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
the enzyme is a flavoprotein
Klebsiella pneumoniae
NADH
the enzyme shows selectivity (V/K ratio of 10) for NADH over NADPH
Klebsiella pneumoniae
NADPH
the enzyme shows selectivity (V/K ratio of 10) for NADH over NADPH
Klebsiella pneumoniae
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.042
-
Urate
pH 8.0, cosubstrate 1.5 mM NADH
Klebsiella pneumoniae
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
45000
-
recombinant enzyme including the affinity tag, determined by SDS-PAGE
Klebsiella pneumoniae
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
additional information
Klebsiella pneumoniae
urate oxidase can catalyze the slow conversion of NADPH to NADP+ in the absence of urate
?
-
-
-
urate + NADH + H+ + O2
Klebsiella pneumoniae
part of the purine catabolic pathway
5-hydroxyisourate + NAD+ + H2O
unstable in aqueous buffer, hydrolyzing spontaneously to 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
enzyme is purified by Ni2+-nitrilotriacetic acid chromatography
Klebsiella pneumoniae
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
urate oxidase can catalyze the slow conversion of NADPH to NADP+ in the absence of urate
696321
Klebsiella pneumoniae
?
-
-
-
-
urate + NADH + H+ + O2
part of the purine catabolic pathway
696321
Klebsiella pneumoniae
5-hydroxyisourate + NAD+ + H2O
unstable in aqueous buffer, hydrolyzing spontaneously to 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline
-
-
?
urate + NADH + H+ + O2
urate hydroxylase shows selectivity (V/K ratio of 10) for NADH over NADPH
696321
Klebsiella pneumoniae
5-hydroxyisourate + NAD+ + H2O
unstable in aqueous buffer, hydrolyzing spontaneously to 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
42
-
Urate
pH 8.0, 1.5 mM NADH as cosubstrate
Klebsiella pneumoniae
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Klebsiella pneumoniae
Other publictions for EC 1.14.13.113
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
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)
724386
Hicks
Structural and mechanistic stu ...
Klebsiella pneumoniae, Klebsiella pneumoniae ATCC 700721
Biochemistry
52
477-487
2013
-
-
1
1
2
-
-
6
-
-
-
2
-
3
-
-
1
-
-
-
-
-
2
1
-
-
-
6
-
-
-
1
-
-
-
-
-
1
1
1
2
-
-
-
-
6
-
-
-
2
-
-
-
1
-
-
-
-
2
1
-
-
-
6
-
-
-
-
-
-
-
-
6
6
727370
Michiel
Microbial urate catabolism: ch ...
Xanthomonas campestris
Environ. Microbiol. Rep.
4
642-647
2012
-
-
1
-
-
-
-
3
-
-
2
2
-
7
-
-
-
-
-
-
-
-
2
1
-
-
-
3
-
-
-
3
-
-
-
-
-
1
3
-
-
-
-
-
-
3
-
-
2
2
-
-
-
-
-
-
-
-
2
1
-
-
-
3
-
-
-
-
-
-
-
-
3
3
696321
O'Leary
Biochemical characterization o ...
Klebsiella pneumoniae
Biochemistry
48
3033-3035
2009
-
-
1
-
-
-
-
1
-
-
1
2
-
2
-
-
1
-
-
-
-
-
3
-
-
-
-
1
1
-
-
3
-
-
-
-
-
1
3
-
-
-
-
-
-
1
-
-
1
2
-
-
-
1
-
-
-
-
3
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
692892
Pope
Purine utilization by Klebsiel ...
Klebsiella oxytoca, Klebsiella oxytoca M5al
J. Bacteriol.
191
1006-1017
2008
-
-
1
-
-
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-