Information on EC 3.2.1.167 - baicalin-beta-D-glucuronidase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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

EC NUMBER
COMMENTARY hide
3.2.1.167
-
RECOMMENDED NAME
GeneOntology No.
baicalin-beta-D-glucuronidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
baicalin + H2O = baicalein + D-glucuronate
show the reaction diagram
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
baicalein degradation (hydrogen peroxide detoxification)
-
-
baicalein metabolism
-
-
wogonin metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
5,6,7-trihydroxyflavone-7-O-beta-D-glucupyranosiduronate glucuronosylhydrolase
The enzyme also hydrolyses wogonin 7-O-beta-D-glucuronide and oroxylin 7-O-beta-D-glucuronide with lower efficiency [4]. Neglegible activity with p-nitrophenyl-beta-D-glucuronide [2].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
subsp. coagulans
-
-
Manually annotated by BRENDA team
Bge
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-nitrophenyl beta-D-glucuronide + H2O
4-nitrophenol + beta-D-glucuronic acid
show the reaction diagram
-
-
-
?
4-nitrophenyl beta-D-glucuronide + H2O
4-nitrophenol + D-glucuronate
show the reaction diagram
5,6,7-trihydroxyflavone-7-beta-D-glucuronide + H2O
5,6,7-trihydroxyflavone + D-glucuronate
show the reaction diagram
-
-
-
-
?
5,6,7-trihydroxyflavone-7-O-beta-D-glucoronate + H2O
5,6,7-trihydroxyflavone + D-glucuronate
show the reaction diagram
-
i.e. baicalin
i.e. baicalein
-
?
5,6,7-trihydroxyflavone-7-O-beta-D-glucoronate + H2O
5,6,7-trihydroxyflavone-7-O-beta-D-glucoronate + D-glucuronate
show the reaction diagram
5,7-dihydroxy-6-methoxyflavone-7-O-beta-D-glucoronate + H2O
5,7-dihydroxy-6-methoxyflavone + D-glucuronate
show the reaction diagram
-
i.e. oroxylin 7-O-beta-D-glucuronide, i.e. 5-hydroxy-6-methoxy-4-oxo-2-phenyl-4H-chromen-7-yl beta-D-glucopyranosiduronic acid
-
-
?
5,7-dihydroxy-8-methoxyflavone-7-O-beta-D-glucoronate + H2O
5,7-dihydroxy-8-methoxyflavone + D-glucuronate
show the reaction diagram
baicalin + H2O
baicalein + D-glucuronate
show the reaction diagram
estrone 3-(beta-D-glucuronide) + H2O
estrone + D-glucuronate
show the reaction diagram
wogonoside + H2O
?
show the reaction diagram
-
-
-
-
?
wogonoside + H2O
wogonin + ?
show the reaction diagram
0.125 mM of wogonoside is completely transformed into wogonin within 3 h at 37°C, pH 5.0
-
-
?
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
baicalin + H2O
baicalein + D-glucuronate
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
Ca2+ and Zn2+ have no significant effect on enzyme activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ag+
-
0.3 mM, 20-25% inhibition
D-glucose
-
10 mM, 15% inhibition
D-glucuronate
-
10 mM, 15% inhibition
Fe3+
-
10 mM: 50% inhibition
additional information
-
2-mercaptoethanol has no influence on activity up to 10 mM
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Mg2+
-
50 mM, 1.2 fold activation
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.154 - 0.156
4-nitrophenyl beta-D-glucuronide
0.0097 - 0.038
5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate
0.19 - 0.36
5,6,7-trihydroxyflavone-7-beta-D-glucuronide
0.0391
5,7-dihydroxy-6-methoxyflavone-7-O-beta-D-glucoronate
-
pH 6.5, 28°C
0.0307
5,7-dihydroxy-8-methoxyflavone-7-O-beta-D-glucoronate
-
pH 6.5, 28°C
1.5 - 2
Baicalin
2.02
wogonoside
-
recombinant His-tagged enzyme, pH 5.0, 37°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
31.8 - 53.7
4-nitrophenyl beta-D-glucuronide
0.011 - 639
5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate
18.5 - 21.3
Baicalin
6.37
wogonoside
Lactobacillus brevis
-
recombinant His-tagged enzyme, pH 5.0, 37°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
204 - 348
4-nitrophenyl beta-D-glucuronide
6225
4.2 - 52810
5,6,7-trihydroxyflavone 7-O-beta-D-glucoronate
5859
10.6 - 14
Baicalin
3678
2.58
estrone 3-(beta-D-glucuronide)
Lactobacillus brevis
-
recombinant His-tagged enzyme, pH 5.0, 37°C
202534
3.15
wogonoside
Lactobacillus brevis
-
recombinant His-tagged enzyme, pH 5.0, 37°C
165071
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.284
at 37°C and pH 5.0, using 4-nitrophenyl-beta-D-glucuronide as substrate
7.57
-
purified native enzyme, pH 5.0, 37°C, substrate baicalin
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.7
-
in 50 mM Na-citrate buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 7
-
pH 3.0: about 50% of maximal activity, pH 7.0: about 65% of maximal activity
3.5 - 7
-
activity range, profile overview
4.5
-
activity completely disappeares below pH 4.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 80
-
activity range, profile overview
40 - 70
-
40°C: about 60% of maximal activity, 70°C: about 50% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.8
-
isoelectric focusing
5.4
-
chromatofocusing
5.5
-
isoelectric focusing
5.52
calculated from sequence
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
55000
-
4 * 55000, SDS-PAGE
56027
x * 56027, calculated from sequence
58400
-
x * 58400, SDS-PAGE
60000
-
x * 60000, native enzyme, SDS-PAGE
71000
recombinant enzyme, SDS-PAGE
200000 - 220000
-
gel filtration
230000
-
gel filtration
290000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homotetramer
tetramer
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3
-
purified enzyme, inactivation
731207
3.5 - 9
-
purified enzyme, stable at
731207
4
-
30 min, about 50% loss of activity
706477
5 - 8
at 37°C, the GUS remains stable for 80 min at pH values ranging from 5.0 to 8.0. Activity decreases quickly above pH 7.0
709499
5 - 5.5
-
30 min, stable
706477
6
-
30 min, about 35% loss of activity
706477
7
-
30 min, about 40% of maximal activity
706477
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 60
-
purified enzyme, stable at
30 - 70
50 - 60
the purified enzyme exhibits a half-life of 1 h at 60°C and more than 2 h at 50°C
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
no appreciable loss in activity is found during 10 repeated reaction cycles of GUS encapsulated in biomimetic alginate/protamine/silica capsules
-
the calcium alginate-encapsulated GUS retains up to 88% of its free-form activity with an encapsulation efficiency of 77%. Conversion of baicalin by free and calcium alginate-encapsulated GUS results in the baicalein productivities of 80% and 65%, respectively. The calcium alginate-encapsulated GUS shows no appreciable loss in activity after four repeated cycles, and 90% of its initial activity remains after 26-day storage at 4°C
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 10 mM NaH2PO4/Na2HPO4 buffer, 1 mM mercaptoethanol, pH 7.0, 50% glycerol, 2 months, 10-20% loss of activity
-
4°C, 10 mM NaH2PO4/Na2HPO4 buffer, 1 mM mercaptoethanol, pH 7.0, 50% glycerol, 2 weeks, complete loss of activity
-
4°C, 30 mM Tris-HCl pH 7.0, 11 days, 26% loss of activity
-
4°C, 30 mM Tris-HCl pH 7.0, 5 days, 19% loss of activity
-
4°C, free enzyme in 30 mM Tris–HCl (pH 7.0), 11 days, about 25% loss of activity
-
4°C, free enzyme in 30 mM Tris–HCl (pH 7.0), 26 days, 96% loss of activity
-
4°C, free enzyme in 30 mM Tris–HCl (pH 7.0), 5 days, about 20% loss of activity
-
4°C, GUS encapsulated in biomimetic alginate/protamine/silica capsules in 30 mM Tris–HCl (pH 7.0), 11 days, no loss of activity
-
4°C, GUS encapsulated in biomimetic alginate/protamine/silica capsules in 30 mM Tris–HCl (pH 7.0), 26 days, 10% loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ni-NTA column chromatography
recombinant His-tagged enzyme from Escherichia coli strain Rosetta 2 (DE3) by nickel affinity chromatography and dialysis, native enzyme 1800fold from cell-free extract by repeated anion exchange chromatography and gel filtration, followed by dialysis
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain Rosetta 2 (DE3)
-
expressed in Escherichia coli strain GMS407 (beta-glucuronidase-deficient)
expression in Escherichia coli. Escherichia coli having pET28a/sGUS shows much higher beta-glucuronidase activity using baicalein 7-O-beta-D-glucuronide as a substrate than Escherichia coli carrying pET28a. The minor enzyme activity observed in the latter is considered to be due to endogenous beta-glucuronidase
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E194A
beta-glucuronidase activity of the mutant enzyme with baicalein 7-O-beta-D-glucuronide is 86% of wild-type activity
E212A
beta-glucuronidase activity of the mutant enzyme with baicalein 7-O-beta-D-glucuronide is 1.2% of wild-type activity
E225A
beta-glucuronidase activity of the mutant enzyme with baicalein 7-O-beta-D-glucuronide is 94% of wild-type activity
E272A
beta-glucuronidase activity of the mutant enzyme with baicalein 7-O-beta-D-glucuronide is 70% of wild-type activity
E329A
beta-glucuronidase activity of the mutant enzyme with baicalein 7-O-beta-D-glucuronide is 0.3% of wild-type activity
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis