Information on EC 1.7.3.3 - factor-independent urate hydroxylase

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

EC NUMBER
COMMENTARY
1.7.3.3
-
RECOMMENDED NAME
GeneOntology No.
factor-independent urate hydroxylase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
urate + O2 + H2O = 5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
urate + O2 + H2O = 5-hydroxyisourate + H2O2
show the reaction diagram
catalytic mechanism, overview
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
oxidation
D0VWQ1
-
oxidation
DQ887577, -
-
redox reaction
-
-
-
-
reduction
-
-
-
-
additional information
-
the enzyme catalyzes the degradation of urate to [S]-allantoin through 5-hydroxyisourate as a metastable intermediate
PATHWAY
KEGG Link
MetaCyc Link
Caffeine metabolism
-
Metabolic pathways
-
Microbial metabolism in diverse environments
-
Purine metabolism
-
urate degradation to allantoin I
-
SYSTEMATIC NAME
IUBMB Comments
urate:oxygen oxidoreductase
This enzyme was previously thought to be a copper protein, but it is now known that the enzymes from soy bean (Glycine max), the mould Aspergillus flavus and Bacillus subtilis contains no copper nor any other transition-metal ion. The 5-hydroxyisourate formed decomposes spontaneously to form allantoin and CO2, although there is an enzyme-catalysed pathway in which EC 3.5.2.17, hydroxyisourate hydrolase, catalyses the first step. The enzyme is different from EC 1.14.13.113 (FAD-dependent urate hydroxylase).
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
AaUO
Q17J02
-
AaUO
Aedes aegypti NH-Rockefeller
Q17J02
-
-
Fasturtec
-
trade name of rasburicase
N-35
-
-
-
-
Nodule specific uricase
-
-
-
-
Nodulin 35
-
-
-
-
Nodulin 35 homolog
-
-
-
-
Non-symbiotic uricase
-
-
-
-
oxidase, urate
-
-
-
-
Rasburicase
-
-
Rasburicase
-
commercially available recombinant enzyme
Rasburicase
-
recombinant Uox
Rasburicase
Q00511
-
Rasburicase
-
recombinant enzyme
Uaz
Q00511
-
Urate oxidase
-
-
-
-
Urate oxidase
Q17J02
-
Urate oxidase
Aedes aegypti NH-Rockefeller
Q17J02
-
-
Urate oxidase
Arthrobacter globiformis FERM BP-360
-
-
-
Urate oxidase
-
-
Urate oxidase
Bacillus subtilis LMD 69.3
-
-
-
Urate oxidase
DQ887577
-
Urate oxidase
-
-
Urate oxidase
-
-
urate oxidoreductase
-
-
urate oxidoreductase
-
-
-
uric acid oxidase
-
-
-
-
uricase
-
-
-
-
uricase
Arthrobacter globiformis FERM BP-360
-
-
-
uricase
Q00511
-
uricase
Bacillus fastidiosus ATCC 26904, Bacillus fastidiosus ATCC 29604
C5HDG5
-
-
uricase
Bacillus subtilis LMD 69.3
-
-
-
uricase
-
-
uricase
-
-
uricase
-
-
uricase
-
-
uricase
P16164
-
Uricoenzyme
-
-
Uricozyme
-
trade name of urate oxidase from Aspergillus flavus
CAS REGISTRY NUMBER
COMMENTARY
9002-12-4
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
NH-Rockefeller strain
UniProt
Manually annotated by BRENDA team
Aedes aegypti NH-Rockefeller
NH-Rockefeller strain
UniProt
Manually annotated by BRENDA team
Arthrobacter globiformis FERM BP-360
FERM BP-360
-
-
Manually annotated by BRENDA team
expressed in Saccharomyces cerevisiae
SwissProt
Manually annotated by BRENDA team
soil isolate
-
-
Manually annotated by BRENDA team
strain ATCC 26904
-
-
Manually annotated by BRENDA team
Bacillus fastidiosus ATCC 26904
-
UniProt
Manually annotated by BRENDA team
Bacillus fastidiosus ATCC 29604
-
UniProt
Manually annotated by BRENDA team
TB-90
-
-
Manually annotated by BRENDA team
subspecies subtilis LMD 69.3
-
-
Manually annotated by BRENDA team
Bacillus subtilis LMD 69.3
subspecies subtilis LMD 69.3
-
-
Manually annotated by BRENDA team
Candida sp.
-
-
-
Manually annotated by BRENDA team
atlantic cod
Uniprot
Manually annotated by BRENDA team
partial sequence
DQ887577
GenBank
Manually annotated by BRENDA team
inoculated with Bradyrhizobium japonicum
-
-
Manually annotated by BRENDA team
thioredoxin fusion protein
-
-
Manually annotated by BRENDA team
Atlantic halibut
Uniprot
Manually annotated by BRENDA team
strain ZZJ4-1
-
-
Manually annotated by BRENDA team
strain ZZJ4-1
-
-
Manually annotated by BRENDA team
cultivar Golden Gate
-
-
Manually annotated by BRENDA team
African lungfish
Uniprot
Manually annotated by BRENDA team
yeast-like fungi
endosymbiont of Nilaparvata lugens
-
-
Manually annotated by BRENDA team
maize
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
Q00511
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
Candida sp.
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-, Q3S561
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
Q3S563, -
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
yeast-like fungi
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-, Q3S560
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
Q00511
-
-
-
-
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
Q9LD80
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-, Q3S562
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-, C5HDG5
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
Q5FZI9
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
P78609
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
conversion to allantoin
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
D0VWQ1
ureide pathway
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
DQ887577, -
ureide pathway
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
enzyme production is induced by addition of uric acid to the culture medium
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
detection of two discrete enzyme-bound intermediates by single-turnover stopped-flow techniques
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
it is proposed that T69 and K9 form a catalytic diad in which K9 deprotonates T69 to allow it to abstract the proton from the N9 position of the substrate to generate the dianion
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
first step in uric acid degradation
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
purine metabolism
a metastable intermediate, which is further degrades to allantoin
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
urate oxidase catalyzes the hydroxylation of uric acid into the metastable product 5-hydroxyisourate in the presence of molecular oxygen as part of the purine degradation pathway. 5-Hydroxyisourate decays slowly to allantoin, a process independent of oxygen and associated with the release of CO
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
dioxygen-binding site structure, overview
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
substrate binding involves residues Arg176 and Glu228, that hold the substrate, Phe159 closing one end of the cavity below, and the two residues Asn254 and Thr57, forming another tweezers above the mean plane of the ligand that construct a location where efficient electron transfer can take place at a low energy level via the catalytic triad Thr57, Lys10, and His256
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
Bacillus fastidiosus ATCC 26904
C5HDG5
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
Bacillus subtilis LMD 69.3
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
Bacillus fastidiosus ATCC 29604
C5HDG5
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
Arthrobacter globiformis FERM BP-360
-
enzyme production is induced by addition of uric acid to the culture medium
-
-
?
uric acid + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
purine degradation
-
-
?
5-hydroxyisourate + O2
(S)-allantoin + H2O2 + CO2
show the reaction diagram
Q00511
-
-
-
?
additional information
?
-
-
uricase can function as a voltage-sensitive channel that is highly selective to urate, relative to K+ and Cl-
-
-
-
additional information
?
-
-
enzyme catalyzes the oxidation of uric acid to a more soluble and easily excreted compound, allantoin
-
-
-
additional information
?
-
-
purine degradation, urate oxidase catalyzes the oxidation of uric acid with poor solubility to produce 5-hydroxyisourate and allantoin
-
-
-
additional information
?
-
-
rasburicase acts at the end of the purine catabolic pathway, and unlike allopurinol, it does not induce accumulation of xanthine or hypoxanthine
-
-
-
additional information
?
-
-
anion-pi interactions are present in the active site of the enzyme and are energetically favorable
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
Candida sp.
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
yeast-like fungi
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
conversion to allantoin
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
D0VWQ1
ureide pathway
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
DQ887577, -
ureide pathway
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
enzyme production is induced by addition of uric acid to the culture medium
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
first step in uric acid degradation
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
purine metabolism
a metastable intermediate, which is further degrades to allantoin
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
urate oxidase catalyzes the hydroxylation of uric acid into the metastable product 5-hydroxyisourate in the presence of molecular oxygen as part of the purine degradation pathway. 5-Hydroxyisourate decays slowly to allantoin, a process independent of oxygen and associated with the release of CO
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
-
-
-
?
urate + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
Arthrobacter globiformis FERM BP-360
-
enzyme production is induced by addition of uric acid to the culture medium
-
-
?
uric acid + O2 + H2O
5-hydroxyisourate + H2O2
show the reaction diagram
-
purine degradation
-
-
?
additional information
?
-
-
enzyme catalyzes the oxidation of uric acid to a more soluble and easily excreted compound, allantoin
-
-
-
additional information
?
-
-
purine degradation, urate oxidase catalyzes the oxidation of uric acid with poor solubility to produce 5-hydroxyisourate and allantoin
-
-
-
additional information
?
-
-
rasburicase acts at the end of the purine catabolic pathway, and unlike allopurinol, it does not induce accumulation of xanthine or hypoxanthine
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
no cofactors
-
additional information
-
no cofactors
-
additional information
D0VWQ1
UOX is unique among the oxygen-requiring enzymes in the sense that it does not need a cofactor to convert uric acid to 5-hydroxyisourate
-
additional information
-
the catalytic mechanism of UOX does not imply any cofactor
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
strong activation at 10 mM
CaCl2
-
enhances activity
Co2+
-
-
copper
-
neither copper nor iron detected
copper
-
enzyme contains 0.15 mol of copper
copper
-
copper : uricase ratio is 1 : 7
copper
-
copper : uricase ratio is 1 : 1
copper
-
negligible amount of copper
copper
-
contains 0.2 mol copper per mol enzyme protein
Cu2+
-
enzyme contains copper, inhibited by excessive addition of Cu2+
Fe3+
-
activates
Iron
-
neither copper nor iron detected
Iron
-
enzyme contains less than 0.05 mol of iron
Iron
-
Fe3+: stimulation
Iron
-
iron : uricase ratio is 1 : 50
Iron
-
contains nearly one mol per mol of enzyme
Iron
-
contains 0.1 mol iron per mol enzyme; Fe3+: stimulation
Iron
-
slight stimulation
K+
-
slight activation at 10 mM
Mg2+
-
strong activation at 10 mM
Na+
-
slight activation at 10 mM
NaCl
-
enhances activity
NH4Cl
-
slight activation at 10 mM
Mn2+
P78609
2 mM, 170% of initial activity
additional information
-
the catalytic mechanism of UOX does not imply any metal ion
additional information
-
a metal ion is possibly strongly bound in the enzyme and forms part of the uricase structure
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1,3,7-Trimethylxanthine
-
i.e. caffeine, slight
2,2'-dipyridyl
-
-
2,2'-dipyridyl
-
weak
2,9-Dimethyl-1,10-phenanthroline
-
neo-cuproin
3,7-Dimethylxanthine
-
i.e. theobromine, slight
3-Methyluric acid
-
-
5,5'-dithiobis(2-nitrobenzoate)
-
-
5-Azaorotate
-
-
5-Azaorotate
-
-
7-Methyluric acid
-
-
8-Azaxanthine
-
potent inhibitor
8-Azaxanthine
D0VWQ1
substrate analogue
8-Azaxanthine
-
competitive inhibitor, binding structure, overview
8-Azaxanthine
-
substrate analogue binds in the active site of the enzyme
8-Azaxanthine
-
-
8-Azaxanthine
-
anion-pi interactions are present in the active site of the enzyme and are energetically favorable. Uric acid and 8-azaxanthine are able to interact favorably with cyanide and chloride ions, respectively and both uric acid and 8-azaxanthine react with water
8-nitroxanthine
-
-
9-methyluric acid
-
-
9-methyluric acid
-
competitive versus urate
9-methyluric acid
-
substrate analogue
adenine
-
weak
adenine
-
negligible inhibition at 0.5 mM at pH 9.2
Ag+
-
95% inhibition at 1 mM
Allantoic acid
-
weak
allantoin
-
weak
Amelide
-
-
-
arginine
-
weak
aspartic acid
-
slight
Ba2+
Candida sp.
-
-
beta-mercaptoethanol
-
about 60% residual activity after 1 h incubation with 0.5 mM beta-mercaptoethanol at pH 8.5 and 25C
Biguanidine salts
-
inactivation is pH-dependent: slightly inhibitory below pH 10, rapid inactivation at high pH
CN-
-
the presence of residue Phe159 enhances the interaction energy of the anion with the urate pi system
Cu2+
Candida sp.
-
-
Cu2+
-
enzyme contains copper, inhibited by excessive addition of Cu2+, 98% inhibition by 1 mM CuCl2, 97% inhibition by 1 mM CuSO4
Cu2+
-
strong inhibition at 0.2 mM, negligible inhibition at 0.005 mM
Cu2+
P78609
2 mM, complete inactivation
Cyanurate
-
-
D-Sorbitol
-
about 70% residual activity after 1 h incubation with 0.5 mM D-sorbitol at pH 8.5 and 25C
Dicyandiamide
-
inactivation is pH-dependent: small below pH 10, rapid increase at high pH
diethyldithiocarbamic acid
-
-
Fe2+
-
strong inhibition at 0.2 mM, negligible inhibition at 0.005 mM
-
Fe3+
Candida sp.
-
-
-
glutamine
-
weak
glyoxylic acid
-
weak
Guanidinium salts
-
inactivation is pH-dependent: slightly inhibitory below pH 10, rapid inactivation at high pH
Hg2+
Candida sp.
-
-
Hg2+
-
98% inhibition at 1 mM
hydroxylamine
-
-
Hydroxypurines
-
-
-
hypoxanthine
-
weak
hypoxanthine
-
weak
hypoxanthine
-
negligible inhibition at 0.5 mM at pH 9.2
inosine 5'-monophosphate
-
-
iodoacetate
-
low effect
Li+
-
18.5% inhibition at 1 mM
N-ethylmaleimide
-
-
N-ethylmaleimide
-
-
o-Iodosobenzoate
-
-
o-phenanthroline
-
-
o-phenanthroline
-
-
o-phenanthroline
-
not
o-phenanthroline
-
weak
o-phenanthroline
-
99% inhibition at 2 mM
Oxopurines
-
-
-
p-chloromercuribenzoate
-
slight effect
p-chloromercuribenzoate
-
-
p-chloromercuribenzoate
-
-
phosphate
-
no inactivation by phosphate, in presence of borate or dithiothreitol
pyrazinoate
-
channel activity completely disappears
Salicylhydroxamic acid
-
-
SDS
-
49% inhibition at 0.5% w/v
Sodium deoxycholate
-
about 90% residual activity after 1 h incubation with 0.5 mM sodium deoxycholate at pH 8.5 and 25C
Thiourea
-
slight
Trichloropurine
-
-
Urate
-
substrate inhibition: above 0.120 mM
Urate
-
above 0.125 mM
xanthine
-
competitive versus urate, noncompetitive versus O2
ZnCl2
-
1 mM, 91% inhibition
additional information
-
no inhibition by 1 mM Fe(ClO4)3 or 1 mM AgNO3
-
additional information
-
production of an egg yolk antibody specific to microbial uricase and its inhibitory effects on uricase activity
-
additional information
-, Q17J02
reduced levels of mRNA levels of urate oxidase after knock down of urate oxidase gene expression
-
additional information
-
no inhibition by Cu2+, Fe3+, or Zn2+
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Uric acid
DQ887577, -
uric acid (0.3%) is an inducer for uricase production, concentrations higher than 0.3% do not enhance the enzyme productivity
additional information
-, Q17J02
1 h after feeding with ammonium chloride highest expression of urate oxidase in fat body, 6 h after feeding with ammonium chloride highest expression of urate oxidase in malpighian tubules; in fat body 3fold increased expression towards end of blood meal digestion; in malpighian tubule expression induced in response to a blood meal, maximum level 24 h after feeding
-
additional information
Candida sp.
-
dithiothreitol treatment leads to an insignificant change in specific activity below 1%
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.031
-
O2
-
-
0.061
-
O2
-
-
0.0000135
-
Urate
-
mutant enzyme K9M, apparent Km in air-saturated buffer
0.00002
-
Urate
-
mutant enzyme F179A, apparent Km in air-saturated buffer
0.00156
-
Urate
-
mutant enzyme F179Y, apparent Km in air-saturated buffer
0.0034
-
Urate
-
wild type enzyme, apparent Km in air-saturated buffer
0.0047
-
Urate
Q9LD80
GST-uricase fusion protein
0.0053
-
Urate
Q9LD80
GST-free recombinant uricase
0.0064
-
Urate
Q5FZI9
mutant A296V, pH 8.6, 25C; mutant R291K/A296V/A301S/K303R, pH 8.6, 25C
0.0065
-
Urate
Q5FZI9
mutant H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25C
0.0085
-
Urate
Q5FZI9
wild-type, pH 8.6, 25C
0.017
-
Urate
Q5FZI9
mutant A89T/G91A7V92M/H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25C
0.028
-
Urate
-
pH 8.0, mutant enzyme D70A
0.0337
-
Urate
P78609
pH 8.0, 37C
0.034
-
Urate
-
pH 8.0, wild-type enzyme
0.0472
-
Urate
-
Rasburicase
0.051
-
Urate
-
pH 8.0, mutant enzyme K9M
0.075
-
Urate
-
pH 7.0, 37C
0.17
-
Urate
-
pH 8.0, mutant enzyme T69A
0.204
-
Urate
-
at pH 9.2
0.27
-
Urate
-
unmodified enzyme, 50 mM borate buffer, 25C, pH 9.2
0.31
-
Urate
-
pH 8.5, 30C
0.547
-
Urate
-
pH 8.0, mutant enzyme T69A/K9M
0.8
-
Urate
-
unmodified enzyme, 50 mM borate buffer, 40C, pH 9.2
1.05
-
Urate
-
pH 8.0, mutant enzyme T69V
0.00588
-
Uric acid
-
-
0.0098
-
Uric acid
-
-
0.01
-
Uric acid
-
-
0.01
-
Uric acid
-
-
0.014
-
Uric acid
-
enzyme without histidine tag
0.015
-
Uric acid
-
enzyme with histidine tag
0.015
-
Uric acid
-
-
0.018
-
Uric acid
-
-
0.021
-
Uric acid
-
-
0.032
-
Uric acid
-
-
0.034
-
Uric acid
-
-
0.035
-
Uric acid
-
-
0.042
-
Uric acid
-
-
0.042
-
Uric acid
-
poly(N-acryloylmorpholine)-OSu-uricase
0.05
-
Uric acid
-
native uricase, monomethoxypoly(ethylene glycol) N-leucine-OSu-uricase and branched monomethoxypoly(ethylene glycol) N-leucine-OSu-uricase
0.099
-
Uric acid
-
-
0.25
-
Uric acid
DQ887577, -
0.1 M sodium borate, pH 8.5 containing 2 mM uric acid, 30 mM 4-aminoantipyrine, 1.5% phenol and peroxidase (15 U/ml), 37C, 20 min
1.5
-
Uric acid
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
3.01
-
Urate
-
pH 8.5, 30C
8.7
-
Urate
Q5FZI9
mutant A89T/G91A7V92M/H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25C
13.6
-
Urate
Q5FZI9
wild-type, pH 8.6, 25C
15.5
-
Urate
Q5FZI9
mutant H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25C
16.1
-
Urate
Q5FZI9
mutant A296V, pH 8.6, 25C
16.8
-
Urate
Q5FZI9
mutant R291K/A296V/A301S/K303R, pH 8.6, 25C
18.1
-
Urate
-
Rasburicase
31.3
-
Uric acid
-
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
510
-
Urate
Q5FZI9
mutant A89T/G91A7V92M/H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25C
17750
1580
-
Urate
Q5FZI9
wild-type, pH 8.6, 25C
17750
2390
-
Urate
Q5FZI9
mutant H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25C
17750
2510
-
Urate
Q5FZI9
mutant A296V, pH 8.6, 25C
17750
2620
-
Urate
Q5FZI9
mutant R291K/A296V/A301S/K303R, pH 8.6, 25C
17750
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.008
-
oxonate
-
unmodified enzyme, 50 mM borate buffer, 25C, pH 9.2
0.041
-
xanthine
-
more than 0.03 mM
0.055
-
xanthine
-
unmodified enzyme, 50 mM borate buffer, 25C, pH 9.2
0.1
-
xanthine
-
unmodified enzyme, 50 mM borate buffer, 40C, pH 9.2
4.5
-
xanthine
-
competitive with urate
0.0021
-
8-nitroxanthine
-
competitive inhibitor versus urate at pH 8.0
additional information
-
additional information
-
1-methylurate, 3-methylurate, 7-methylurate do not inhibit significantly
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.067
-
-
wild-type, pH 8.9, 25C
2.49
-
-
urate oxidase including p-azido-L-phenylalanine instead of Phe at position 281, in 0.1 M borate, pH 8.4
2.67
-
-
recombinant strain overexpressing the enzyme, pH 8.9, 25C
2.99
-
Q5FZI9
mutant A89T/G91A7V92M/H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25C
3.6
-
-
-
3.74
-
-
cotyledons, 4 d old
4.22
-
Q5FZI9
wild-type, pH 8.6, 25C
4.91
-
Q5FZI9
mutant H245L/E252A/M253I/R291K/A296V/A301S/K303R, pH 8.6, 25C
5.29
-
-
-
5.32
-
-
purified enzyme
5.68
-
Q5FZI9
mutant A296V, pH 8.6, 25C
5.94
-
Q5FZI9
mutant R291K/A296V/A301S/K303R, pH 8.6, 25C
6.85
-
-
-
8.26
-
-
urate oxidase including p-azido-L-phenylalanine instead of Phe at position 170, in 0.1 M borate, pH 8.4
9.35
-
-
male rats
10.5
-
-
in the cell lysate
12.2
-
Candida sp.
-
-
13.3
-
-
hypocotyls, 4 d old
15.3
-
-
female
15.4
-
-
roots, 4 d old
16.33
-
-
natural uricase, in 0.1 M borate, pH 8.4
18
-
-
crude extract, pH 8.0, 37C
21.5
-
-
after DEAE Sepharose FF chromatography
25.7
-
-
after Phenyl-Sepharose FF chromatography
27
-
-
after HiLoad 26/60 Superdex 75 gel filtration
38.4
-
P78609
recombinant protein, pH 8.0, 37C
39
-
-
after 2.1fold purification, pH 8.0, 37C
1790
-
-
nodules, 21 d old
additional information
-
-
-
additional information
-
-
7.1 U/ml predicted optimum activity
additional information
-
-
among the parameters investigated in shaking flask cultures, the pH value of medium and inoculum size has great influence on the recombinant uricase production, the maximum extracellular uricase yield of 2.6 U/ml is obtained in shaking flask culture; at pH 5.5, the extracellular uricase production reaches top of 7.5 U/ml at 58 h, when fermentation is performed at pH 6.5 for 62 h, 14.5 U/ml of extracellular uricase and 23.3 U/ml of intracellular uricase are produced, the total specific uricase production at pH 6.5 is 1.7times of that at pH 5.5; in high density fermentation in YPG medium at 37C, extracellular uricase activity increases significantly during the first 40 h, highest extracellular uricase level of 52.3 U/ml is obtained after 58 h of induction, as well as the intracellular activity of 60.3 U/ml, after 86 h of fermentation and 58 h of induction, a total uricase activity of 112600 U/l is obtained, the extracellular and intracellular yields of uricase in high cell density fermentation increased by 3.7fold and 3.5fold compared with the batch fermentation; the combined use of fed-batch culture and pH-controlled strategy increases the expression level of uricase significantly, the extracellular uricase production of 52.3 U/ml (approximately 2.1 g/l of protein) is obtained, which is much higher than that produced by recombinant Escherichia coli strains
additional information
-
-
rasburicase causes enzymatic degradation of uric acid within blood, plasma and serum samples at room temperature, the genetic absence of this molecule in humans and its proteic nature together with poor accuracy in purifi cation and a slow production process confer a high immunogenicity to the compound, leading to elevated rate of hypersensivity reactions; rasburicase does not interact with allopurinol, cytarabine, methylprednisolone, methotrexate, mercaptopurine, thioguanine, etoposide, daunorubicin, cyclophosphamide, or vincristine; rasburicase maintains the same mechanism of action as the non-recombinant form of urate oxidase, but simply shows a significantly lower reaction rate; repeated use of rasburicase increases risk of hypersensitivity reactions: skin rashes (1.4%), urticaria, bronchospasm (1%), dyspnea, hypoxemia, and anaphylactic shock (1%)
additional information
-
DQ887577, -
enzyme activity is 0.06 U/ml at pH 4.0; enzyme activity is 0.09 U/ml at pH 4.5; enzyme activity is 0.11 U/ml at pH 5.0; enzyme activity is 0.21 U/ml with potassium nitrate as nitrogen source; enzyme activity is 0.22 U/ml at pH 10.0; enzyme activity is 0.31 U/ml with ammonium sulfate as nitrogen source; enzyme activity is 0.32 U/ml with sodium glutamate as nitrogen source; enzyme activity is 0.37 U/ml at pH 5.5; enzyme activity is 0.42 U/ml at pH 9.5; enzyme activity is 0.45 U/ml with citric acid as carbon source; enzyme activity is 0.46 U/ml with glucose as carbon source; enzyme activity is 0.46 U/ml with peptone as nitrogen source; enzyme activity is 0.47 U/ml at pH 6.0; enzyme activity is 0.47 U/ml with lactose as carbon source; enzyme activity is 0.49 U/ml with starch as carbon source; enzyme activity is 0.57 U/ml at pH 9.0; enzyme activity is 0.57 U/ml with soybean flour as nitrogen source; enzyme activity is 0.63 U/ml at pH 8.5; enzyme activity is 0.65 U/ml with beef extract as nitrogen source; enzyme activity is 0.67 U/ml with sucrose as carbon source; enzyme activity is 0.69 U/ml with yeast extract as nitrogen source; enzyme activity is 0.72 U/ml at pH 8.0; enzyme activity is 0.78 U/ml with maize milk as nitrogen source; enzyme activity is 0.98 U/ml at pH 6.5; enzyme activity is 1.00 U/ml at pH 7.5; enzyme activity is 1.25 U/ml at pH 7.0; when the strain is cultured at 30C at pH 7.0 for 30-36 h with of 0.6% corn steep liquor as nitrogen source, the uricase activity peaks at 1.25 U/ml
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
-
-
yield of recombinant uricase is significantly improved by the combined use of a high cell-density cultivation technique and a pH control strategy of switching culture pH from 5.5 to 6.5 in the induction phase
7
-
DQ887577, -
optimal pH for uricase production in culture flasks
8
8.5
-
borate and phosphate buffer
8
-
-
isoforms UI to UIV
8.4
-
-
assay at
8.5
-
Candida sp.
-
-
8.5
-
-
-
8.5
-
-
assay at
8.5
-
DQ887577, -
assay at
8.5
-
P78609
-
8.6
-
-
free enzyme
8.9
-
-
assay at
9
-
Q9LD80
-
9.2
-
-
the unmodified uricase has a pH optimum of slightly below pH 9.2 which is not altered by modification with NHS esters of monomethoxy-poly(ethylene glycol)-5000 or monomethoxy-poly(ethylene glycol)-350
9.5
-
-
-
9.5
-
-
glycine buffer
9.5
-
-
immobilized enzyme
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
11
-
pH 6.5: about 40% of maximal activity, pH 10.0: about 50% of maximal activity
6.5
9.5
-
free uricase shows at least 50% relative activity between pH 6.5 and 9.5, around pH 7.5, free uricase remains 81.16% of its maximum activity, while the uricase loaded in the lipid vesicles remains almost the same high activity (178.26%) as its optimum activity (179.72%)
7
11
-
pH 7: about 50% of activity maximum, pH 11: about 40% of activity maximum
7.4
9.6
-
50% of activity maximum at pH 7.4 and pH 9.6, free enzyme
8
10.2
-
pH 8.0: about 35% of activity maximum, pH 10.2: about 55% of activity maximum
8
11
-
50% of activity maximum at pH 8 and pH 11, immobilized enzyme
8.5
10.5
-
50% of activity maximum at pH 8.5 and pH 10.5
8.5
9.5
Candida sp.
-
-
8.6
9.5
-
-
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
assay at
27
-
-
isoform UIV
30
-
Candida sp.
-
-
35
-
-
5 min incubation test
37
-
-
isoforms UI, UII, UIII
40
-
-
-
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
50
-
20C: about 60% of activity maximum; 50C: about 50% of activity maximum
20
50
-
20C: about 70% of activity maximum; 50C: about 60% of activity maximum
20
55
-
20C: about 80% of maximal activity, maximal activity at 30C, 55C: about 55% of maximal activity
20
70
-
free uricase shows at least 50% relative activity between 20 and 70C
35
60
-
-
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.96
-
-
isoelectric focusing, pH gradient 3.5-9.5
7.6
-
-
isoelectric focusing, rasburicase
8.46
-
-
determined by means of P/ACE 5000
9.38
-
Q9LD80
calculation from nucleotide sequence
additional information
-
-
isoelectric focusing presents a trail of pI values between 6.55 and 7.6
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Q3S563, -
-
Manually annotated by BRENDA team
-
strain CGMCC 2.120 is used as source of uricase gene
Manually annotated by BRENDA team
-
wild-type and PEX5-defective CHO cell lines each stable producing the enzyme
Manually annotated by BRENDA team
Candida sp.
-
-
Manually annotated by BRENDA team
Aedes aegypti NH-Rockefeller
-
-
-
Manually annotated by BRENDA team
-
activity increases once the haustorium has differentiated after Uromyces phaseoli infection. A up-regulation of urate oxidase gene expression occurs at the post-transcriptional level rather than an overexpression of the urate oxidase gene. A general pathogenic effect and host urate oxidase, and purine pool depauperation
Manually annotated by BRENDA team
-
no activity in liver of New World monkeys, low activity in liver of Old World monkeys
Manually annotated by BRENDA team
Q3S563, -
-
Manually annotated by BRENDA team
Aedes aegypti NH-Rockefeller
-
-
-
Manually annotated by BRENDA team
-
no uricase is detected in mycelium grown in minimal medium containing NH4Cl as sole nitrogen source. Uricase activity is increased 10fold to 40fold under derepression conditions and is induced by exogenous uric acid
Manually annotated by BRENDA team
Q9LD80
nodule
Manually annotated by BRENDA team
additional information
Q3S563, -
not in skeletal muscle or kidney
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
isoforms UI, UIII, UIII
-
Manually annotated by BRENDA team
Arthrobacter globiformis FERM BP-360
-
-
-
Manually annotated by BRENDA team
-
the enzyme is mainly localized in the membrane of PEX5-defective mutant cells
Manually annotated by BRENDA team
-
limited to large peroxisomes in uninfected cells of root nodules of Glycine max inoculated with Bradyrhizobium japonicum
Manually annotated by BRENDA team
-
the enzyme is mainly localized in the peroxisomal matrix of wild-type cells
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Bacillus sp. (strain TB-90)
Canis familiaris
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
32000
35000
-
SDS-PAGE, gel filtration
32880
-
yeast-like fungi
-
deduced from nucleotide sequence of cDNA
33270
-
-
determination of nucleotide sequence of cDNA and calculation of corresponding amino acid sequence
34000
-
-
SDS-PAGE
34000
-
-
SDS-PAGE
34000
-
-
recombinant uricase determined by SDS-PAGE
35050
-
-
deduced from nucleotide sequence of cDNA
35060
-
-
MALDI-TOF mass spectrometry
35780
-
-
MALDI-TOF mass spectrometry
35790
-
-
calculated from amino acid sequence
50000
-
-
gel filtration
58680
-
-
MALDI-TOF mass spectrometry
58900
-
-
calculated from amino acid sequence
60000
-
-
about 60000 Da, SDS-PAGE
68000
-
-
gel filtration
70000
76000
Candida sp.
-
gel filtration
100000
-
-
gel filtration
100000
-
-
gel filtration
102000
-
-
polyacrylamide disc electrophoresis
105000
-
-
gel filtration
109000
-
-
gel filtration
114000
128000
-
PAGE, gel filtration
115000
123000
-
PAGE, gel filtration
117000
-
-
gel filtration
120000
122000
-
PAGE, gel filtration
120000
122000
-
-
120000
140000
-
PAGE, gel filtration
120000
-
-
gel filtration, equilibrium sedimentation
125000
-
-
short-column meniscus depletion sedimentation equilibrium
125000
-
-
PAGE
128000
-
-
PAGE
128000
-
-
-
136300
141600
-
ultracentrifugation
137000
-
-
for the homotetramer, determined by neutron crystallographic analysis
140000
-
Candida sp.
-
gel filtration
144000
-
-
native enzyme, gel filtration
145000
150000
-
gel filtration
151000
-
-
Sephadex G-200 gel filtration
230000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 34000, SDS-PAGE
?
-
x * 33000, SDS-PAGE
?
-
x * 62000, rasburicase, SDS-PAGE
?
-
x * 98000, uricase-MBP fusion protein
?
-
x * 34000, SDS-PAGE
?
P78609
x * 34160, MALDI-TOF
?
-
x * 39700, isoform UI, x * 3050, isoform UII, x * 55300, isoform UIII, x * 18000, isoform UIV, SDS-PAGE
?
-
x * 34000, SDS-PAGE
-
dimer
-
at pH 8.8 the enzyme associates to form a dimer of about 68000
dimer
-
at pH 8.8 the enzyme associates to form a dimer of about 10000
heterotetramer
-
alpha2beta2, 2 * 36000 + 2 * 39000, SDS-PAGE
heterotetramer
-
alpha2beta2, 2 * 35000 + 2 * 37000, SDS-PAGE
hexamer
-
alpha6, 6 * 37000, SDS-PAGE
homotetramer
-
4 * 33750, X-ray crystallography
homotetramer
D0VWQ1
determined by X-ray diffraction measurements, the AgUOX-native structure is solved by molecular replacement using the program AmoRe, pairs of dimers are stacked face-to-face to form a tetramer
homotetramer
-
determined by X-ray crystallography, formed by crystallographic symmetry operations
homotetramer
-
determined by gel filtration, tunnel-shaped homotetramer, each of the 4 active sites in UOX is formed by residues from 2 subunits and located at the subunit-subunit interface
homotetramer
-
x-ray crystallography
homotetramer
-
4 * 000, SDS-PAGE, homotetrameric uricase in water dissociates into inactive homodimers that can form active homotetramers again in solutions of high ionic strength
homotetramer
Bacillus fastidiosus ATCC 26904
-
4 * 000, SDS-PAGE, homotetrameric uricase in water dissociates into inactive homodimers that can form active homotetramers again in solutions of high ionic strength
-
monomer
-
native enzyme at pH 7.5 is a monomer with MW 32000-35000
monomer
-
native enzyme at pH 7.5 is a monomer with MW 50000
monomer
Candida sp.
-
1 * 70000-76000, SDS-PAGE
monomer
-
1 * 64000
tetramer
-
alpha4, 4 * 30000, SDS-PAGE
tetramer
-
alpha4, 4 * 33000, SDS-PAGE
tetramer
-
alpha4, 4 * 32000, SDS-PAGE
tetramer
-
alpha4, 4 * 35052, calculated from deduced amino acid sequence
tetramer
-
alpha4, 4 * 32000-34000, SDS-PAGE
tetramer
-
-
tetramer
-
4 * 33000, SDS-PAGE; 4 * 33858, calculation from nucleotide sequence
tetramer
-
4 * 36000, SDS-PAGE and matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry
tetramer
Candida sp.
-
4 * 36000, SDS-PAGE
tetramer
Arthrobacter globiformis FERM BP-360
-
4 * 33000, SDS-PAGE; 4 * 33858, calculation from nucleotide sequence
-
trimer
-
alpha3, 3 * 32000, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystallizations are performed using the hanging-drop vapour-diffusion method at 19.9C, structures of crystals soaked with the substrate uric acid, the inhibitor 8-azaxanthin and allantoin are determined at 1.9-2.2 A resolution, 2 homotetramers comprise the asymmetric crystallographic unit, each subunit contains 2 T-fold domains of topology, which are usually found in purine- and pterin-binding enzymes, the uric acid substrate is bound tightly to the enzyme by interactions with Arg180, Leu222 and Gln223 from one subunit and with Thr67 and sp68 of the neighbouring subunit in the tetramer
D0VWQ1
; Rasburicase
-
crystallization of large proteins in the presence of polyethylene glycol
-
crystals of about a few tens of micrometres in size, which is nucleated previously in crystallization batch containing 5% PEG 8000, 100 mM NaCl, 8 mg/ml uox-substrate complex and 100 mM Tris-HCl pH 8.5, are used as seeds and their size and quality are further improved using a temperature-control device, large crystals of Uox, co-crystallized with its substrates analogues 8-azaxanthine, 9-methyluric acid or the natural substrate in the presence of cyanide (0.5-2 mg/ml), and soaks with the natural substrate in the absence of cyanide, diffracting to high resolutions are obtained, in the presence of different inhibitors, the crystal form of Uox has a body-centred orthorhombic symmetry and one of the largest primitive unit-cell volumes (a: 80 A, b: 96 A, c: 106 A)
-
enzyme in complex with substrate urate and inhibitor cyanide, X-ray diffraction structure determination and analysis
-
hanging drop vapour diffusion method
-
ligand-free Uox crystallized with NH4Cl and 15% (w/v) PEG 8000, ligand-free Uox crystallized in water with 10% (w/v) PEG 8000, ligand-free Uox crystallized with NaCl and 15% (w/v) PEG 8000, ligand-free Uox crystallized with (NH4)2SO4 and 15% (w/v) PEG 8000, ligand-free Uox crystallized with NaCl and 8% PEG 8000, ligand-free Uox crystallized with KCl and 10% (w/v) PEG 8000, and Uox complexed with 8-azaxanthine and crystallized with NaCl and 10% (w/v) PEG 8000, in 50 mM Tris buffer pH 8.0
-
recombinant enzyme in complex with inhibitor 8-azaxanthine in presence of O2 or Cl-, batch technique at room temperature, 10-15 mg/ml protein with an excess of 0.5-2 mg/ml of 8-azaxanthin in 50 mM Tris/HCl, pH 8.5, in the presence of 5-8% w/v PEG 8000 and 0.05 M NaCl, 24-48 h, X-ray diffraction structure determination and analysis at 1.6-1.7 A resolution
-
sitting drop vapour diffusion method
-
sitting drop vapour diffusion method using buffered D2O
-
sitting-drop vapour-diffusion method at room temperature
-
sitting-drop vapour-difusion method. Four different crystal forms of Uox are analyzed. In the presence of uracil and 5,6-diaminouracil crystals usually belong to the trigonal space group P3(1)21, the asymmetric unit of which contains one tetramer of Uox. Chemical oxidation of 5,6-diaminouracil within the protein may occur, leading to the canonical (I222) packing with one subunit per asymmetric unit. Coexistence of two crystal forms, P2(1) with two tetramers per asymmetric unit and I222, is found in the same crystallization drop containing another inhibitor, guanine. A fourth form, P2(1)2(1)2 with one tetramer per asymmetric unit, results in the presence of cymelarsan, an additive
-
homology modeling of monomeric enzyme. The highly conserved residue Gly290 could interact with Asn262 and His264. Residue substitutions near Gly290 may affect its spatial orientation and result in changes in catalysis.Gly290 is likely to participate in the structure of the active site and to be involved in oxygen-binding
Q5FZI9
to 1.93 A resolution. Space group P212121 with unit cell parameters a 69.16 A, b 139.31 A, c 256.33 A, and alpha =beta =gamma =90
P78609
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
11
-
35C, 1 h, stable
6
-
-
45C, 30 min, 40% loss of activity
6
-
-
25C, 18 h, about 60% loss of activity
6.5
10.5
-
22C, 30 min, immobilized enzyme, stable
7
10
-
purified enzyme, 25C, 18 h, stable
7
11
-
10 min, stable
7
9.5
-
in case of uricase entrapped in lipid vesicles, the remaining activity keeps more than 90% during the pH of 7.0-9.5, and the maximum remaining activity is 98.04% at pH 8.0 when incubated at 40C or 40 min. For the free uricase, the maximum remaining activity is 86.59% at pH 8.5 when incubated at 40C or 40 min
7
-
-
UOX is deactivated at different protein concentrations at 45C in 20 mM phosphate containing 0.15 M NaCl, 0.01 mg/ml UOX is deactivated much faster than its counterparts at concentrations of 0.1 and 1.0 mg/ml
8
11
-
4C, 60 days, 35% loss of activity
8
9
-
45C, 30 min, stable
10
-
-
45C, 30 min, 30% loss of activity
11
-
-
25C, 18 h, about 60% loss of activity
12
-
-
22C, 30 min, 55% loss of activity
additional information
-
-
uricase activity in 50 h culture broth with pH values of 5.5 and 6.0 decreases more rapidly than that in cultures with pH values of 6.5 and 7.0, at pH 5.5, about 78% of initial uricase activity is lost within 25 h, under the same conditions, more than 85% of initial uricase activity remains in culture broth of pH 6.5 and 7.0, uricase activity in 66 h culture broth with pH 7.0 degrades much more rapidly than that in samples from 50 h culture, while for pH 6.5, the uricase is still stable, loss of uricase activity is caused by the degradation in acidic environment by proteases secreted by the host cells or releases from host cell lyses, low pH may cause instability of uricase
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
10
80
-
about 80% relative activity at 10C, about 85% relative activity at 20-30C, about 90% relative activity at 37C, about 40% relative activity at 40C, about 30% relative activity at 50C, about 20% relative activity at 60C, about 10% relative activity at 70C, and no activity at 80C after 30 min incubation
18
20
-
2 days, 70% loss of activity
40
-
-
30 min, stable below
40
-
-
below pH 9.5, 2 h, very slow loss of activity in absence or presence of Cu2+
40
-
-
10 min, stable below
40
-
-
at 40C in sodium borate buffer at pH 9.2, the unmodified uricase shows a thermo-inactivation half-life of about 40 h. Modification of the uricase by monomethoxy-poly(ethylene glycol)-350 slightly enhances its thermostability, and modification by monomethoxy-poly(ethylene glycol)-5000 increases its thermo-inactivation half-life to over 85 h at 40C in sodium borate buffer at pH 9.2
45
-
Candida sp.
-
after 5 min 50% of initial activity
50
-
-
pH 7.8, 10 min, stable below
50
-
-
10 min, stable
52
-
-
uricase covalently linked to monomethoxypoly(ethylene glycol) N-leucine-OSu-uricase and branched monomethoxypoly(ethylene glycol) N-leucine-OSu-uricase 50% loss of activity
55
-
-
uricase entrapped in lipid vesicles at enzyme concentrations of 0.005 and 0.1 mg/ml shows no loss of activity after 5 h at 55C. Uricase entrapped in lipid vesicles at enzyme concentrations of 0.01 mg/ml shows about 35% loss of activity after 5 h at 55C. Uricase entrapped in lipid vesicles at enzyme concentrations of 0.005 mg/ml shows about 50% loss of activity after 5 h at 55C. The free uricase at 0.005 mg/ml is rapidly deactivated to about 30% of the initial activity within an incubation time of 2 h, while more than 70% of the initial enzyme activity remains for the uricase at 0.1 mg/ml in the identical incubation time
60
61
-
native uricase and poly(N-acryloylmorpholine)-OSu-uricase 50% loss of activity
60
-
-
pH 7.8, 10 min, 50% loss of activity
60
-
-
30 min, about 40% loss of activity
60
-
-
10 min, 55% loss of activity
60
-
-
10 min, stable
65
-
-
30 min, the purified enzyme retains 98.9% of its original activity
70
-
-
30 min, the purified enzyme retains 64% of its original activity
75
-
DQ887577, -
enzyme is thermostable, after heat treatment at 75C for 45 min, the uricase retains about 100% of its initial activity
80
-
DQ887577, -
70% of initial activity remains after 45 min at 80C
100
-
-
35% relative activity at 100C
additional information
-
-
thermal inactivation rises steeply as CuSO4 concentration rises from 0.025 to 0.175 mM and as the pH of the medium exceedes 9.5
additional information
-
-
thermal deactivation of recombinant UOX at neutral pH is associated with the loss of intersubunit hydrogen bonds, subunit is unstable at room temperature and unfolds rapidly at 100C, tetramer has significantly higher stability than its subunit
additional information
-
-
the molecular structure of enzyme has a reversible change at a temperature between 30C and 60C
additional information
-
Candida sp.
-
the main phase of thermal inactivation follows an irreversible two-state mechanism, with loss of about 20% of the helical structure, loss of the majority of the tertiary structure, and partial exposure of tryptophan residues to solution being approximately concurrent with activity loss. This process results in the formation of aggregated molten globules. In addition, a rapid reversible denaturation phase occurs that is not completely coupled to the main phase. Enzyme inactivation is inhibited by the presence of glycerol and trimethylamine oxide. NaCl destabilizes the enzyme at elevated temperature
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ammonium sulfate protects from inactivation at low pH
-
Cu2+ inactivates at low temperature, uric acid prevents inactivation
-
Fe3+ partially protects against inactivation at low pH or at low ionic strength, stimulates reactivation
-
proteolytic digestion by endopeptidases cause rapid loss of activity, exopeptidases have slight effect
-
after modification with monomethoxy-poly(ethylene glycol)-5000, the recombinant intracellular uricase shows residual activity of about 65%
-
conjugated uricases are more stable to trypsin digestion
-
proteolytic digestion by endopeptidases cause rapid loss of activity, exopeptidases have slight effect
-
uricase is reversibly inactivated in solutions of low ionic strength (like during dialysis against water). After incubation for 2 h in 100 mM sodium chloride in water at 4C, the dialysis-inactivated uricase shows about 70% of the maximal specific activity. After incubation for 2 h in 100 mM Tris-HCl pH 8.0 plus 100 mM NaCl at 4C, the dialysis-inactivated uricase shows about 90% of the maximal specific activity. After pre-incubation for 0.5 h in sodim borate plus 100 mM NACL at 25C followed by the direct addition of urate to measure its activity, the dialysis-inactivated uricase shows about 80% of the maximal specific activity
-
borate stabilizes
-
dithiothreitol effect of treatment with dithiothreitol on extraction and purification
-
dithiothreitol stabilization
-
EDTA stabilizes
-
at the physiological pH, significant increase of enzyme activity is found for the uricase entrapped in the lipid vesicles (1.8times that of free uricase) at their respective optimum pH. Free uricase shows rapid decrease in its enzymatic activity with a half life of less than 20 min when incubated with trypsin. Uricase entrapped in the lipid vesicles gradually loses its activity but still 50% of the original activity remains after 60 min (remaining activity is 7.32% in case of free uricase)
-
dithiothreitol prevents polymerization and stabilizes throughout purification
-
little loss of activity by freeze-drying
-
lower stability in solutions of phosphate buffer than in borate buffer
-
proteolytic digestion by endopeptidases cause rapid loss of activity, exopeptidases have slight effect
-
Repeated freezing and thawing has no effect
-
urate oxidase from female rat livers is more stable than enzyme from male rat livers
-
urea, 4 M, several h without loss of activity
-
borate stabilizes
-
EDTA stabilizes
-
stability of immobilized enzyme depends on the time of stirring during immobilization and on the quantity of enzyme used
-
unusually resistant to guanidinium chloride
-
unusually resistant to SDS
-
frozen enzyme retains complete activity
-
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
DMSO
-
the residual recombinant UOX activity in the presence of DMSO is significantly higher than that in pure H2O, the residual UOX activity increases in response to the increase in the DMSO concentration up to 20%, further increase in DMSO concentration (50-70%) results in significant UOX deactivation
Methanol
-
the residual recombinant UOX activity in the presence of methanol is significantly higher than that in pure H2O, the residual UOX activity increases in response to the increase in the methanol concentration up to 20%, further increase in methanol concentration (50-70%) results in significant UOX deactivation
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
sensitive to O2
-
246662
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-15C, 100 mM borate buffer, pH 8.0, containing 100 mM or more ammonium sulfate
-
-12C, 0.10 mol/l Tris/HCl buffer (pH 8.0), 8 months, less than 20% loss of activity, enzyme remains stable
-
0-4C, crystals in (NH4)2SO4 solution, 3 months, stable
-
4C, 0.10 mol/l Tris/HCl buffer (pH 8.0), 10 weeks, less than 20% loss of activity
-
-20C, purified enzyme, 72 h, 20% loss of activity
-
37C, purified enzyme, 12 h and 48 h, 50% and 100% loss of activity
-
4C, purified enzyme, 72 h, 20% loss of activity
-
-20C, 72 h, no loss of activity
P78609
4C, 72 h, no loss of activity
P78609
4C, free uricase in borate buffer (pH 8.5), 28 days, 70% loss of activity
-
4C, uricase in lipid vesicles in borate buffer (pH 8.5), 28 days, no loss of activity
-
4C, several weeks
-
3C, 0.15% sodium carbonate, several weeks, stable
-
4C, 20 mM phosphate buffer, pH 7.8, for at least 1 month
-
4C, 7 days, 40% loss of activity
-
4C, 100 mM Tris-HCl buffer, pH 9.5, 90% of activity after 5 days
yeast-like fungi
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
by anion exchange and hydropbobic interaction chromatography
D0VWQ1
DEAE Sepharose FF chromatography, Phenyl-Sepharose FF chromatography and HiLoad 26/60 Superdex 75 gel filtration
-
recombinant enzyme
-
Superdex S200PG gel filtration
-
DEAE-cellulose column chromatography
-
enzyme synthesized with C-terminal 6-histidine tag
-
Ni-NTA column chromatography
-
uricase-MBP fusion protein
-
-
Candida sp.
-
ammonium sulfate precipitation, ion exchange chromatography and gel filtration
-
by Ni-NTA affinity chromatography
-
native enzyme 19.7fold by ammonium sulfate fractionation, anion exchange exchange and hydrophobic interaction chromatography, and gel filtration
-
-
yeast-like fungi
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
a plasmid containing the AgUOX gene is introduced into the expression host cell Escherichia coli DH1
D0VWQ1
expression in Escherichia coli, Escherichia coli harboring pUOD1 produces 20fold higher uricase than the original Arthrobacter strain, even without an inducer
-
expressed in Escherichia coli BL21 (DE3)
-
expressed in Escherichia coli BL21 (DE3) cells
-
expressed in Saccharomyces cerevisiae
-
expression in Saccharomyces cerevisiae
-
expressed in Escherichia coli BL21(DE3) cells
-
overexpression in Escherichia coli JM109
-
expressed in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) cells
-
expression in Escherichia coli
Candida sp.
-
expression in Escherichia coli
Q5FZI9
complete cDNA and genomic DNA fragment coding for urate oxidase is isolated and characterized, heterologous expression in Escherichia coli
Q9LD80
developing of a method for genetically incorporating p-azido-L-phenylalanine into target protein in Escherichia coli in a site-specific manner utilizing a tyrosyl suppressor tRNA/aminoacyl-tRNA synthetase system, substitution of p-azido-L-phenylalanine for F170 or F281 in urate oxidase, optimization of the system by adding a Shine-Dalgarno sequence and tandem suppressor tRNA in order to increase the expression levels of tyrosyl suppressor tRNA and aminoacyl-tRNA synthetase
-
expression in Escherichia coli
P78609
uricase production by the recombinant Hansenula polymorpha strain MU200 harboring Candida utilis uricase gene under the control of methanol oxidase promoter using Saccharomyces cerevisiae alpha-factor signal peptide as the secretory sequence
-
expression in Escherichia coli RR1
-
expression in Escherichia coli
-
expression in NM 538
-
overexpression in Pichia angusta
-
expression in Escherichia coli MC1061
-
expressed in Chinese hamster ovary cells
-
using the recombinant DNA technique, enzyme is obtained from a genetically modified Saccharomyces cerevisiae strain that expresses urate oxidase cDNA, cloned from a strain of Aspergillus flavus
-
expression in pPUO1
-
expression in Escherichia coli DH5alpha
yeast-like fungi
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D79A
-
the KM-value for urate is 82% of the wild-type value
F179A
-
decreases Vmax by 2 orders of magnitude
F179Y
-
mutation decreases Vmax by 2-fold
K9M
-
mutant enzyme has a maximal velocity of 0.4% of the wild-type value. Ionization at pH 6.4 that is observed with the wild-type enzyme is absent in the mutant. The KM-value for urate is 1.5fold higher than that of the wild-type enzyme
K9M
-
mutant is not able to generate the dianion of urate and is decreased in activity by over 200-fold
T69A
-
mutant enzyme has a maximal velocity of 3% of the wild-type value. Ionization at pH 6.4 that is observed with the wild-type enzyme is absent in the mutant. The KM-value for urate is 5fold higher than that of the wild-type enzyme
T69A/K9M
-
the KM-value for urate is 16.1fold higher than that of the wild-type enzyme
T69V
-
the KM-value for urate is 30.9fold higher than that of the wild-type enzyme
A296V
Q5FZI9
about 50% ioncrease in catalytic efficiency
A89T/G91A7V92M/H245L/E252A/M253I/R291K/A296V/A301S/K303R
Q5FZI9
replacement with the corresponding residues of human enzyme. About 30% of wild-type catalytic efficiency
H245L/E252A/M253I/R291K/A296V/A301S/K303R
Q5FZI9
replacement with the corresponding residues of human enzyme. Catalytic efficiency is higher than in wild-type, but below the efficiency of mutant R291K/A296V/A301S/K303R
I115V/H119R/L120F/H245L/E252A/M253I/R291K/A296V/A301S/K303R
Q5FZI9
replacement with the corresponding residues of human enzyme, complete loss of activity
R119H
Q5FZI9
complete loss of activity
additional information
-
uricase covalently linked to monomethoxypoly(ethylene glycol) N-leucine-OSu, branched monomethoxypoly(ethylene glycol) N-leucine-OSu or poly(N-acryloylmorpholine)-OSu last longer in blood
R291K/A296V/A301S/K303R
Q5FZI9
replacement with the corresponding residues of human enzyme. About 50% increase in catalytic efficiency
additional information
-
thioredoxin urate oxidase fusion protein
additional information
P16164
the recombinantly expressed porcine enzyme, with a C-terminal sequence from baboon uricase, is applicated to patients with refractory gout, due to a mutation in the uricase gene, by intravenous injection in a PEG-bound form, pharmacokinetics and pharmacodynamics, safety, and efficacy of the treatment in a clinical trial, overview
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
analysis
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a colorimetric 96-well microtiter plate assay for the determination of urate oxidase activity and its kinetic parameters based on hydrogen peroxide quantitation. The general advantages of the colorimetric assay are easy handling of large amounts of samples at the same time, the possibility of automation, and the need for less material
medicine
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urate oxidase is used to reduce toxic urate accumulation during chemotherapy
medicine
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substitute for allopurinol in the management of gout and hyperuricaemia
medicine
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determining the urate concentration in blood and urine
medicine
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determining the urate concentration in blood and urine
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analysis
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modified colorimetric assay for uricase activity in flexible 96-well microtiter plates using the uricase/uric acid/horseradish peroxidase/4-aminoantipyrine/3,5-dichloro-2-hydroxybenzene sulfonate colorimetric reaction. The method is much more efficient than the conventional ones and greatly reduces assay time from 4 days to less than 20 h
medicine
Candida sp.
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determining the urate concentration in blood and urine
drug development
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urate oxidase has the potential to be a therapeutic target for the treatment of gout
medicine
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uricase is an important medical enzyme which can be used to determine urate in clinical analysis, to therapy gout, hyperuricemia, and tumor lysis syndrome
synthesis
P78609
high-yield expression of uricase in Escherichia coli and establishment of an efficient three-step protein purification protocol. The purity of the recombinant protein is more than 98% and the specific activity is 38.4 IU/mg
medicine
DQ887577, -
determination of the urate concentration in blood and urine is required for the diagnosis of gout as urate accumulation is a causative factor of gout in humans
analysis
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development of an urate-selective microbial biosensor cells of the recombinant thermotolerant methylotrophic yeast Hansenula polymorpha as biorecognition element. The UOX producing cells are coupled to horseradish peroxidase and immobilized on graphite electrodes by physical entrapment behind a dialysis membrane. A high urate selectivity with a detection limit of about 8 microM is found
medicine
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treatment od tumor lysis syndrome, recombinant urate oxidase is effective in reducing uric acid and preventing uric acid accumulation in patients with hematologic malignancies with hyperuricemia or at high risk of developing it, rasburicase represents an effective alternative to allopurinol to promptly reduce uric acid levels, improve patients electrolyte status, and reverse renal insufficiency
medicine
P16164
the enzyme might be useful in the treatment of patients with refractory gout, overview