Information on EC 3.2.1.139 - alpha-glucuronidase

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

EC NUMBER
COMMENTARY hide
3.2.1.139
-
RECOMMENDED NAME
GeneOntology No.
alpha-glucuronidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
an alpha-D-glucuronoside + H2O = an alcohol + D-glucuronate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
hydrolysis of O-glycosyl bond
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
alpha-D-glucosiduronate glucuronohydrolase
Considerable differences in the specificities of the enzymes from different fungi for alpha-D-glucosiduronates have been reported. Activity is also found in the snail.
CAS REGISTRY NUMBER
COMMENTARY hide
37259-81-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
strain 5-16
-
-
Manually annotated by BRENDA team
NRRL Y-2311-1
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
strain J-37
-
-
Manually annotated by BRENDA team
strain J-37
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
Clostridium stercorarium
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
no activity in Neocallimastix frontalis
-
-
-
Manually annotated by BRENDA team
no activity in Neocallimastix patriciarum
-
-
-
Manually annotated by BRENDA team
strain JDR-2
-
-
Manually annotated by BRENDA team
strain TH501b
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Polyporus versicolor
-
-
-
Manually annotated by BRENDA team
strain CBS 814.70
-
-
Manually annotated by BRENDA team
strain CBS 814.70
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain JW/SL-YS485
-
-
Manually annotated by BRENDA team
strain JW/SL-YS485
-
-
Manually annotated by BRENDA team
Tyromyces palustris
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
physiological function
-
alpha-glucuronidase enzymes play an essential role in the full enzymatic hydrolysis of hemicellulose
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,2-linked glucuronic acid of non-reducing xylose-oligosaccahrides + H2O
D-glucuronic acid + ?
show the reaction diagram
2-O-alpha-(4-O-methyl-alpha-D-glucopyranosyluronic)-D-xylobiose + H2O
4-O-methylglucuronic acid + D-xylobiose
show the reaction diagram
-
i.e. aldotriouronic acid
-
-
?
2-O-alpha-(4-O-methyl-alpha-D-glucuronosyl)-xylotriose + H2O
4-O-methyl-alpha-D-glucuronic acid + xylotriose
show the reaction diagram
2-O-alpha-D-glucopyranosyluronic acid-D-xylose + H2O
alpha-D-glucopyranosyluronic acid + D-xylose
show the reaction diagram
-
-
-
-
-
2-O-alpha-D-glucuronosyl-D-xylose + H2O
D-glucuronate + D-xylose
show the reaction diagram
-
-
-
-
-
3-O-alpha-D-glucuronosyl-D-xylose + H2O
D-glucuronate + D-xylose
show the reaction diagram
-
-
-
-
-
4-nitrophenyl 2-(4-O-methyl-alpha-D-glucopyranosyl)-beta-D-xylopyranoside + H2O
4-nitrophenyl beta-D-xylopyranoside + 4-O-methyl-alpha-D-glucopyranose
show the reaction diagram
4-nitrophenyl-2-O-(4-O-methyl-alpha-D-glucuronosyl)-beta-D-xylopyranoside + H2O
4-nitrophenyl-D-xylopyranoside + 4-O-methyl-alpha-D-glucuronate
show the reaction diagram
-
-
-
-
?
4-O-alpha-D-glucuronosyl-D-xylose + H2O
D-glucuronate + D-xylose
show the reaction diagram
-
-
-
-
-
4-O-Me-GlcAalpha(1-2)Xylbeta(1-4)Xylbeta(1-4)Xyl + H2O
4-O-methyl-alpha-D-glucuronic acid + Xylbeta(1-4)Xylbeta(1-4)Xyl
show the reaction diagram
from beechwood, de-estrification of the methyl ester of MeGlcA with glucuronoyl esterase
de-estrification of the methyl ester of MeGlcA with glucuronoyl esterase
-
?
4-O-Me-GlcAalpha(1-2)Xylbeta(1-4)Xylbeta(1-4)[4-O-Me-GlcAalpha(1-2)]Xylbeta(1-4)Xyl + H2O
4-O-methyl-alpha-D-glucuronic acid + Xylbeta(1-4)Xylbeta(1-4)[4-O-Me-GlcAalpha(1-2)]Xylbeta(1-4)Xyl
show the reaction diagram
hydrolysis product of endoxylanases on glucuronoxylan
de-estrification of the methyl ester of MeGlcA with glucuronoyl esterase
-
?
4-O-methyl-alpha-D-glucopyranuronosyl-(1-2)-beta-D-xylopyranosyl-(1-4)-beta-D-xylopyranose + H2O
4-O-methyl-alpha-D-glucuronic acid + beta-D-xylopyranose
show the reaction diagram
4-O-methyl-alpha-D-glucopyranuronosyl-(1-2)-beta-D-xylopyranosyl-(1-4)-beta-D-xylopyranosyl-(1-4)-beta-D-xylopyranose + H2O
4-O-methyl-alpha-D-glucuronic acid + beta-D-xylopyranosyl-(1-4)-beta-D-xylopyranosyl-(1-4)-beta-D-xylopyranose
show the reaction diagram
4-O-methyl-D-glucuronoxylan + H2O
4-O-methyl-D-glucuronic acid + ?
show the reaction diagram
-
-
-
?
4-O-methyl-glucuronosyl-xylan + H2O
4-O-methyl-glucuronate + xylan
show the reaction diagram
4-O-methyl-glucuronosyl-xylitol + H2O
4-O-methyl-glucuronate + xylitol
show the reaction diagram
4-O-methyl-glucuronosyl-xylobiose + H2O
4-O-methyl-glucuronate + xylobiose
show the reaction diagram
4-O-methyl-glucuronosyl-xyloheptaose + H2O
4-O-methyl-glucuronate + xyloheptaose
show the reaction diagram
-
-
-
-
-
4-O-methyl-glucuronosyl-xylohexaose + H2O
4-O-methyl-glucuronate + xylohexaose
show the reaction diagram
4-O-methyl-glucuronosyl-xylopentaose + H2O
4-O-methyl-glucuronate + xylopentaose
show the reaction diagram
4-O-methyl-glucuronosyl-xylose + H2O
4-O-methyl-glucuronate + xylose
show the reaction diagram
4-O-methyl-glucuronosyl-xylotetraose + H2O
4-O-methyl-glucuronate + xylotetraose
show the reaction diagram
4-O-methyl-glucuronosyl-xylotriose + H2O
4-O-methyl-glucuronate + xylotriose
show the reaction diagram
acetylated glucuronoxylan + H2O
?
show the reaction diagram
-
-
-
-
-
aldobiouronic acid + H2O
?
show the reaction diagram
aldobiouroniuc acid + H2O
?
show the reaction diagram
aldohexaouronic acid + H2O
?
show the reaction diagram
aldopentaouronic acid + H2O
4-O-methyl-D-glucuronic acid + Xylbeta(1-4)Xylbeta(1-4)Xylbeta(1-4)Xyl 2
show the reaction diagram
-
-
-
-
?
aldopentaouronic acid + H2O
?
show the reaction diagram
aldopentauronic acid + H2O
4-O-methyl-D-glucuronic acid + Xylbeta(1-4)Xylbeta(1-4)Xylbeta(1-4)Xyl
show the reaction diagram
aldotetraouronic acid + H2O
?
show the reaction diagram
aldotetrauronic acid + H2O
?
show the reaction diagram
aldotriouronic acid + H2O
?
show the reaction diagram
aldouronic acid + H2O
D-xylose + ?
show the reaction diagram
amylouronate + H2O
D-glucuronate + ?
show the reaction diagram
beechwood glucuronoxylan + H2O
?
show the reaction diagram
beechwood xylan + H2O
4-O-methyl-D-glucuronic acid + D-glucuronic acid
show the reaction diagram
-
-
-
?
beechwood xylan + H2O
?
show the reaction diagram
-
-
-
?
benzyl 4-O-alpha-D-glucuronosyl-beta-D-glucoside + H2O
?
show the reaction diagram
-
-
-
-
-
beta-D-glucuronyl alpha-D-glucuronic acid + H2O
D-glucuronate + alpha-D-glucuronic acid
show the reaction diagram
birchwood xylan + H2O
4-O-methyl-D-glucuronic acid + D-glucuronic acid
show the reaction diagram
-
-
-
?
birchwood xylan + H2O
?
show the reaction diagram
GlcAalpha(1-2)Xylbeta(1-4)Xylbeta(1-4)[GlcAalpha(1-2)]Xylbeta(1-4)Xyl + H2O
alpha-D-glucuronic acid + Xylbeta(1-4)Xylbeta(1-4)[GlcAalpha(1-2)]Xylbeta(1-4)Xyl
show the reaction diagram
generated from the action of endoxylanases on polysaccharides
de-estrification of the methyl ester of MeGlcA with glucuronoyl esterase
-
?
glucuronosyl-xylotriose + H2O
glucuronate + xylotriose
show the reaction diagram
glucuronoxylan + H2O
4-O-methyl-D-glucuronic acid + ?
show the reaction diagram
glycyrrhizin + H2O
glycyrrhetinic acid + 2 D-glucuronate
show the reaction diagram
O-alpha-D-glucopyranosyluronic acid-alpha-D-glucopyranosiduronic acid + H2O
alpha-D-glucopyranosyluronic acid + alpha-D-glucopyranosiduronic acid
show the reaction diagram
-
-
-
-
-
O-alpha-D-glucosyluronic acid alpha-D-glucosiduronic acid
D-glucuronic acid
show the reaction diagram
-
the immobilized enzyme completely hydrolyzes 100 mM of the substrate
-
-
?
O-alpha-D-glucosyluronic acid alpha-D-glucosiduronic acid + H2O
alpha-D-glucosiduronic acid + beta-D-glucosiduronic acid
show the reaction diagram
-
-
alpha-D-glucosiduronic acid and beta-D-glucosiduronic acid are produced in an equimolar ratio
-
?
O-alpha-glucuronosyl alpha-D-glucuronide + H2O
D-glucuronate
show the reaction diagram
-
-
-
-
-
p-nitrophenyl alpha-D-glucuronide + H2O
p-nitrophenol + D-glucuronate
show the reaction diagram
-
-
-
-
-
p-nitrophenyl alpha-D-glucuronopyranoside + H2O
p-nitrophenol + D-glucuronate
show the reaction diagram
-
-
-
?
p-nitrophenyl-alpha-D-glucuronopyranoside + H2O
p-nitrophenol + D-glucuronate
show the reaction diagram
-
-
-
-
?
p-nitrophenyl-beta-D-glucuronide + H2O
p-nitrophenol + D-glucuronate
show the reaction diagram
reduced aldopentaouronic acid + H2O
4-O-methyl-D-glucuronic acid + ?
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
1,2-linked glucuronic acid of non-reducing xylose-oligosaccahrides + H2O
D-glucuronic acid + ?
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ag+
-
2 mM, slight activation
BaCl2
-
slight activation
Ca2+
-
2 mM, slight activation
CaCl2
-
slight activation
Cu2+
-
2 mM, slight activation
Fe3+
-
1 mM, 25% activation
FeCl3
-
slight activation
K+
-
2 mM, slight activation
Mg2+
-
2 mM, slight activation
Na+
-
2 mM, slight activation
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-O-Methyl-D-glucuronic acid
-
-
Ca2+
-
14% inhibition at 1 mM
Cd2+
-
1 mM, strong inhibition
D-glucose
D-glucuronic acid
DTT
-
50 mM, required for maximal activity
K+
-
1 mM, 15% inhibition
Monoiodoacetic acid
N-bromosuccinimide
Ni2+
-
1 mM, 10% inhibition
p-hydroxymercuribenzoate
-
-
PCMB
-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
maximal activation at 150 mM
L-cysteine
maximal activation at 0.6 mM, 15% of the activation with 2-mercaptoethanol
NAD+
maximal activation at 0.5 mM
solka floc
-
low levels of activity
xylan
-
induces activity, maximum activity on beechwood xylan, activity also on oat spelt and birchwood xylan, low levels of activity on wheat arabinoxylan and larchwood xylan
additional information
-
glutaraldehyde stabilizes
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.12
(3beta)-30-hydroxy-11,30-dioxoolean-12-en-3-yl 2-O-beta-D-glucopyranuronosyl-alpha-D-glucopyranosiduronic acid
-
-
17.6
2-O-(4-methyl-alpha-D-glucopyranosyluronic acid)-D-xylose
-
-
0.78
2-O-alpha-(4-O-methyl-alpha-D-glucopyranosyluronic)-D-xylobiose
-
40C, pH 6.5
0.33
2-O-alpha-D-Glucopyranosyluronic acid-D-xylose
-
-
0.21 - 0.58
4-nitrophenyl 2-(4-O-methyl-alpha-D-glucopyranosyl)-beta-D-xylopyranoside
0.19 - 2.1
4-nitrophenyl-2-O-(4-O-methyl-alpha-D-glucuronosyl)-beta-D-xylopyranoside
0.76 - 0.95
4-O-Methyl-glucuronosyl-xylobiose
0.145 - 0.47
4-O-Methyl-glucuronosyl-xylotriose
0.53
aldobiouronic acid
-
-
0.42
aldopentaouronic acid
-
-
0.2 - 0.36
aldotetraouronic acid
0.28
aldotriouronic acid
-
-
0.4 - 0.9
beechwood xylan
0.3 - 1.1
birchwood xylan
0.77 - 0.82
glucuronosyl-xylotriose
0.36
O-alpha-D-glucopyranosyluronic acid-alpha-D-glucopyranosiduronic acid
-
-
-
0.13
p-nitrophenyl alpha-D-glucopyranosyluronic acid
-
-
0.1
p-nitrophenyl alpha-D-glucuronopyranoside
-
1
p-nitrophenyl-alpha-D-glucuronopyranoside
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.014 - 220
4-nitrophenyl-2-O-(4-O-methyl-alpha-D-glucuronosyl)-beta-D-xylopyranoside
69.2
aldobiouronic acid
Cellvibrio japonicus
-
-
131.9
aldopentaouronic acid
Cellvibrio japonicus
-
-
54.9 - 102.5
aldotetraouronic acid
96
aldotriouronic acid
Cellvibrio japonicus
-
-
11.6 - 13.95
beechwood xylan
11.08 - 14.37
birchwood xylan
0.28
p-nitrophenyl alpha-D-glucuronopyranoside
Thermotoga maritima
P96105
-
0.0018
p-nitrophenyl-alpha-D-glucuronopyranoside
Thermotoga maritima
-
-
additional information
additional information
Thermoanaerobacterium sp.
-
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
15.5 - 119
beechwood xylan
13.06 - 36.95
birchwood xylan
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.54
4-O-methyl-glucuronic acid
-
37C, pH 7.0, wild-type enzyme
77.1
glucose
-
37C, pH 7.0, wild-type enzyme
3.84 - 78.3
glucuronic acid
69.5
xylose
-
37C, pH 7.0, wild-type enzyme
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.07
-
crude extract
0.29
-
recombinant His-tagged enzyme from Pichia pastoris, pH 5.0, 28C
1.09
-
fungus native culture supernatant, pH 5.0, 28C
1.647
Clostridium stercorarium
-
-
2.8
-
after 41.5fold purification
11.7
-
purified recombinant enzyme, pH 8.0, 45C
15.3
-
hydrolysis of 2-O-alpha-(4-O-methyl-alpha-D-glucopyranosyluronic)-D-xylobiose
61
-
partially purified enzyme
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2 - 4
-
pH 2: about 30% of maximal activity, pH 4: about 70% of maximal activity
3 - 5
-
pH 3.0: about 80% of maximal activity, pH 5.0: about 50% of maximal activity
3 - 5.8
-
about 45% of maximal activity at pH 3 and at pH 5.8
3.3 - 7
-
pH 3.3: about 60% of maximal activity, pH 7: about 50% of maximal activity
4 - 5.6
-
about 50% of maximal activity at pH 4.5 and 5.6
4 - 5.5
at pH 4.0 25.5% and at pH 5.5 51.6% activity
4.5 - 7.5
-
about 40% of maximal activity at pH 4.5 and pH 7.5
4.8 - 6.3
-
pH 5.8: about 80% of maximal activity, pH 6.3: about 50% of maximal activity
5 - 6.2
-
pH 5: about 85% of maximal activity, pH 6.2: about 65% of maximal activity
5 - 8
-
pH 5: about 35% of maximal activity, pH 8: about 45% of maximal activity
5.5 - 9.5
-
activity range, profile overview
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35
-
mutant enzyme W328E/R329T/R665N
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 60
-
20C: about 35% of maximal activity, 60C: about 60% of maximal activity
20 - 55
-
20C: about 50% of maximal activity, 55C: about 40% of maximal activity, mutant enzyme W328E/R329T/R665N
30 - 60
40 - 75
-
40C: about 35% of maximal activity, 75C: about 30% of maximal activity
45 - 70
-
45C: about 35% of maximal activity, 70C: about 50% of maximal activity
45 - 65
-
45C: about 80% of maximal activity, 65C: about 85% of maximal activity
60 - 70
-
40C: about 40% of maximal activity, 70C: about 90% of maximal activity, wild-type enzyme
60 - 100
-
about 45% of maximal activity at 60C and at 100C
60 - 90
60C: about 50% of maximal activity, 90C: about 80% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
isoelectrofocusing on Multiphor II system using SEVALYT PRECOTES gels and markers from 3-10
4.6
calculated from sequence
5.34
-
calculated from sequence
6.1
-
sequence analysis
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
of the enzyme grown in cultures containing filter paper, barley straw, birchwood xylan or birchwood sawdust as carbon source
Manually annotated by BRENDA team
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
91600
-
sequence analysis
91730
-
recombinant mature enzyme, sequence calculation
100000 - 300000
-
MW ranging from 100000 to 300000, the fraction with the highest activity has a MW of 103000, gel filtration
115000
-
native AUH-I, gel filtration
118000
120000
124000
Clostridium stercorarium
-
gel filtration
130000
-
gel filtration, native PAGE
150000
160000
-
gel filtration
161000
-
gel filtration
180000
-
gel filtration
199000
analytical ultracentrifugation, recombinant wild-type enzyme
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterodimer
homodimer
2 * 85000, about, sequence calculation
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant His6-tagged wild-type and selenomethionine-labeled enzyme or enzyme complexed with alpha-D-glucuronic acid, 10 mg/ml protein mixed with 19% PEG3350, 0.2 M sodium citrate, pH 5.5, soaking with 300 mM glucuronic acid for the complexed structure, use of mother liquor supplemented with 15% v/v PEG 400 or paratone N oil as cryoprotectant, X-ray diffraction structure determination and analysis at 2.14-3.0 A resolution
crystal structure of mutant E292A in complex with its substrate aldobiouronic acid
-
hanging-drop vapor diffusion method
-
hanging drop method, several high resolution crystal structures of the alpha-glucuronidase in complex with its substrate and products: structure of wild-type enzyme, structure of mutant enzyme E285N, mutant enzyme in complex with aldotetraouronic acid
-
hanging-drop vapor diffusion method. Two crystal forms: T1 and M1. T1 form: space group P4(1)2(1)2 or P4(3)2(1)2 with unit-cell dimensions a = b = 76.1 A, c = 331.2 A. The crystals are mechanically strong, are stable in the X-ray beam and diffract X-rays to better than 2.4 A resolution. M1 form: space group P2(1) with unit-cell dimensions a = 65.8, b = 127.4, c = 96.6 A and beta = 97.9. The crystals are quite strong and stable and diffract to better than 2.8 A resolution
-
sitting drop vapor diffusion method, using 20% (w/v) PEG 2000 MME and 0.1 M Tris pH 7.0
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2 - 4
-
immobilized TreDCase
680401
2 - 4.5
-
30C, stable
664379
3 - 7
-
30C, 2 h, stable
26106
3.5
-
50C, in absence of substrate, inactivated in 2 h
26105
4 - 9
-
almost no enzyme activity appears at pHs lower than 4.0 and higher than 9.0
703044
4 - 12
30 min, stable between pH 4 and pH 12
664349
4.5 - 7
-
50C, 1 h, stable
26100
4.8 - 5.5
-
40C, 24 h, stable
26109
5
-
AguA exhibits 85% of its maximal activity at pH 5.0
701796
5 - 11
-
room temperature, 30 min, stable
664926
6.2 - 7.9
-
60C, stable for 1 h
26108
6.6
-
40C, 24 h, 22% loss of activity
26109
11.5
-
room temperature, 30 min, 50% loss of activity
664926
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10 - 40
-
AUH-I is stable at 10-40C, but starts to lose the activity above 40C
30
-
pH 4.0, 24 h, stable below
45
-
4 h, 20% loss of activity
55
rapid inactivation above
55 - 75
-
partial inactivation, but thereafter the residual activity remains nearly constant for several days
62
-
half-life: 1 h
73
-
melting temperature of mutant enzyme W328E/R329T/R665N is 72.9C, melting temperature of wild-type enzyme is 73.4C
73.4
-
denaturation at
80
30 min, 10% loss of activity
90
30 min, complete inactivation
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
susceptible to proteinase attack, inactivation by trypsin is similar to the inactivation rate for the intracellular enzyme malate dehydrogenase
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-18C, 4 months, complete inactivation
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 enzym forms: CM-I and CM-II
-
HiTrap column chromatography
-
Ni-NTA agarose column chromatography, DEAE-Sepharose column chromatography, and Superose 12 gel filtration
partially purified by gel filtration
-
recombinant
recombinant enzyme
-
recombinant enzyme from Saccharomyces cerevisiae strain MH1000pbk by ultrafiltration
-
recombinant His-tagged enzyme from Pichia pastoris by nickel affinity chromatography
-
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) to homogeneity by immobilized metal ion affinity chromatography and gel filtration
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3)pLysE by immobilized metal affinity chromatography and gell filtration
-
soluble phase onto HisTrap HP column, eluted with imidazole gradient in sodium phosphate buffer, pH 8, and 300 mM sodium chloride
supernatant concentrated on Amicon 10 kDa cut-off membranes, anion-exchange chromatography on HiTrap DEAE-FF column with NaCl gradient in 50 mM sodium-phosphate buffer, pH 7.0, fractions between 0.2 and 0.25 M NaCl pooled, concentrated, desalted, equilibrated in 50 mM acetate buffer, pH 4.0, with 2 M (NH4)2SO4, subjected to hydrophobic interaction chromatography on Butyl-FF column, eluted with (NH4)2SO4 gradient, active fractions between 1.1 and 0.61 M (NH4)2SO4 pooled, desalted, concentrated, subjected to 2 additional anion-exchange chromatography steps on Tricorn MonoQ 5/50GL column, first equilibrated with 50 mM sodium acetate buffer, pH 4.0, eluted with NaCl gradient, then equilibrated with 50 mM sodium phosphate buffer, pH 7.0 and NaCl gradient, concentration with Microcon 10 kDa cut-off membrane filter
Superose 12 gel filtration
Toyopearl SuperQ column chromatography, Toyopearl phenyl column chromatography, Resource Q column chromatography, and TSKgel G3000SWXL gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Escherichia coli BL21(DE3)pLysE, Luria-Bertani broth, 37°C, plasmid pET29-Gste-AG-his-S
expressed in Escherichia coli Rosetta(DE3) cells
-
expressed in Escherichia coli Tuner (DE3) cells
expression in Escherichia coli
expression in Saccharomyces cerevisiae
-
from strain A1163, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Pichia pastoris
-
gene deg75-AG, DNA and amino acid sequence determination and analysis, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3)pLysE
-
into pGEM-T Easy vector, plasmids maintained in Escherichia coli JM109
-
overexpression in Escherichia coli using the T7 polymerase expression system
-
Pichia stipitis CBS 6054 grown in YNB medium, cells washed and suspended in YNB medium with a xylooligosaccharide misture and methyl-beta-xylopyranoside, 30°C, 24 h, shaker
recombinant expression in Saccharomyces cerevisiae strain MH1000pbk, optimisation and scale-up of enzyme production in aerobic fed-batch culture with constant growth rate, strain growth parameters at at 30C and pH 5.5, and growth and production profiles , overview. The enzyme is secreted into the culture supernatant
-
recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3)
the gene is isolated from a rumen metagenomic library
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D192A
site-directed mutagenesis, the mutation has no effect on the enzyme activity
D206A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
D332A
site-directed mutagenesis, inactive mutant
D396N
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
D478A
site-directed mutagenesis, the mutation has no effect on the enzyme activity
E162A
site-directed mutagenesis, the mutation has no effect on the enzyme activity
E375A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
E782A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
E785A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
H275A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
H275A/H422A
site-directed mutagenesis, inactive mutant
H422A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
K374A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
N205A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
N398A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
N462A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
R328A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
W169A
site-directed mutagenesis, the mutation has no effect on the enzyme activity
W249A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Y373A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Y420A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
Y425A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Y788A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Y792A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
D365A
-
activity is 0.0001% of wild-type activity
D365C
-
activity is 0.0001% of wild-type activity
E292C
-
activity is 0.0001% of wild-type activity
E393A
-
activity is 0.0001% of wild-type activity
E393C
-
activity is22% of wild-type activity
K288A
-
kcat/KM for the substrate 4-nitrophenyl-2-O-(4-O-methyl-alpha-D-glucuronosyl)-beta-D-xylopyranoside is 0.003% of wild-type value
K360A
-
kcat/KM for the substrate 4-nitrophenyl-2-O-(4-O-methyl-alpha-D-glucuronosyl)-beta-D-xylopyranoside is 0.019% of wild-type value
R325A
-
kcat/KM for the substrate 4-nitrophenyl-2-O-(4-O-methyl-alpha-D-glucuronosyl)-beta-D-xylopyranoside is 0.000096% of wild-type value
V210A
-
kcat/KM for the substrate 4-nitrophenyl-2-O-(4-O-methyl-alpha-D-glucuronosyl)-beta-D-xylopyranoside is 38% of wild-type value
V210G
-
kcat/KM for the substrate 4-nitrophenyl-2-O-(4-O-methyl-alpha-D-glucuronosyl)-beta-D-xylopyranoside is identical to wild-type value
V210N
-
kcat/KM for the substrate 4-nitrophenyl-2-O-(4-O-methyl-alpha-D-glucuronosyl)-beta-D-xylopyranoside is 9.4% of wild-type value
V210S
-
kcat/KM for the substrate 4-nitrophenyl-2-O-(4-O-methyl-alpha-D-glucuronosyl)-beta-D-xylopyranoside is 4.3% of wild-type value
W160A
-
kcat/KM for the substrate 4-nitrophenyl-2-O-(4-O-methyl-alpha-D-glucuronosyl)-beta-D-xylopyranoside is 0.0064% of wild-type value
W543A
-
kcat/KM for the substrate 4-nitrophenyl-2-O-(4-O-methyl-alpha-D-glucuronosyl)-beta-D-xylopyranoside is 0.069% of wild-type value
D274A
-
as active as wild-type enzyme
D364A
-
0.000015% of wild-type activity
D364A/E392C
-
0.0000004% of wild-type activity
E158N
-
0.07% of wild-type activity
E285N
-
0.0002% of wild-type activity
E386Q
-
0.07% of wild-type activity
E392C
-
0.00002% of wild-type activity
E510A
-
20% of wild-type activity
W328E/R329T
-
activity of the monomeric mutant enzyme is significantly lower than activity of dimeric wild-type enzyme
W328E/R329T/R665N
-
activity of the monomeric mutant enzyme is significantly lower than activity of dimeric wild-type enzyme, melting temperature is 0.5C lower than. OPtimal temperature is around 35C, compared to 65 for the wild-type enzym
D274A
-
as active as wild-type enzyme
-
D364A
-
0.000015% of wild-type activity
-
E158N
-
0.07% of wild-type activity
-
E510A
-
20% of wild-type activity
-
W328E/R329T
-
activity of the monomeric mutant enzyme is significantly lower than activity of dimeric wild-type enzyme
-
W328E/R329T/R665N
-
activity of the monomeric mutant enzyme is significantly lower than activity of dimeric wild-type enzyme, melting temperature is 0.5C lower than. OPtimal temperature is around 35C, compared to 65 for the wild-type enzym
-
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
molecular biology
synthesis
additional information
-
immobilized enzyme is effective in high yield production of D-glucuronic acid
Show AA Sequence (250 entries)
Please use the Sequence Search for a certain query.