Information on EC 3.2.1.84 - glucan 1,3-alpha-glucosidase

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

EC NUMBER
COMMENTARY hide
3.2.1.84
-
RECOMMENDED NAME
GeneOntology No.
glucan 1,3-alpha-glucosidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of terminal (1->3)-alpha-D-glucosidic links in (1->3)-alpha-D-glucans
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
hydrolysis of O-glycosyl bond
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Metabolic pathways
-
-
N-Glycan biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
3-alpha-D-glucan 3-glucohydrolase
Does not act on nigeran.
CAS REGISTRY NUMBER
COMMENTARY hide
9073-99-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain RIB40
-
-
Manually annotated by BRENDA team
one isoform
-
-
Manually annotated by BRENDA team
soybean
-
-
Manually annotated by BRENDA team
strain MP-1
-
-
Manually annotated by BRENDA team
strain RM1
E16590
GenBank
Manually annotated by BRENDA team
sequence was found to be identical to previously published data; strain CECT2413
-
-
Manually annotated by BRENDA team
strain F-340, rich in alpha-(1-3)-glucans
-
-
Manually annotated by BRENDA team
strain Rut-C30
-
-
Manually annotated by BRENDA team
strain Rut-C30
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
mung bean
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
loss-of-function in the Arabidopsis thaliana glucosidaseII beta-subunit gene confers an polypeptide elf18-insensitive phenotype
metabolism
-
glucosidase-II has a promiscuous activity as a broad specificity hexosidase
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-methylumbelliferyl-alpha-D-glucopyranoside + H2O
4-methylumbelliferol + alpha-D-glucopyranose
show the reaction diagram
-
high affinity
-
-
?
4-methylumbelliferyl-alpha-D-glucopyranoside + H2O
4-methylumbelliferone + alpha-D-glucose
show the reaction diagram
4-methylumbelliferyl-alpha-D-glucoside + H2O
4-methylumbelliferone + alpha-D-glucose
show the reaction diagram
-
-
-
-
?
4-nitrophenyl alpha-D-glucopyranoside + H2O
4-nitrophenol + alpha-D-glucopyranose
show the reaction diagram
-
-
-
-
?
4-nitrophenyl alpha-D-glucopyranoside + H2O
4-nitrophenol + D-glucopyranose
show the reaction diagram
4-nitrophenyl alpha-D-glucopyranoside + H2O
4-nitrophenol + D-glucose
show the reaction diagram
alpha-1,3-glucan + H2O
alpha-D-glucose
show the reaction diagram
alpha-1,3-glucans + H2O
alpha-D-glucose
show the reaction diagram
-
exo-wise hydrolysis from non-reducing ends
-
-
?
alpha-1,3-mutan + H2O
alpha-D-glucose
show the reaction diagram
beta-glucan:chitin + H2O
?
show the reaction diagram
Glc2Man7GlcNAc2 + H2O
GlcMan7GlcNAc2 + D-glucose
show the reaction diagram
-
-
-
-
?
Glc2Man9GlcNAc2 (G2M9)-protein + H2O
?
show the reaction diagram
-
glucosidase II plays a key role in glycoprotein processing in the endoplasmic reticulum. This enzyme trims two alpha-1,3-linked glucose residues, Glcalpha1,3Glc (cleavage 1) and Glcalpha1,3Man (cleavage 2) from high-mannose type Glc2Man9GlcNAc2 (G2M9)-proteins. A crowded milieu that contains bovine serum albumin greatly enhances the second trimming step (cleavage 2), which deglucosylates Glc1Man9GlcNAc2, but not the first trimming step (cleavage 1), which removes the terminal glucose residue from Glc2Man9GlcNAc2
-
-
?
Glc2Man9GlcNAc2 + H2O
GlcMan9GlcNAc2 + D-glucopyranose
show the reaction diagram
Glc2Man9GlcNAc2 + H2O
GlcMan9GlcNAc2 + D-glucose
show the reaction diagram
-
-
-
-
?
Glc2Man9GlcNAc2 + H2O
Man9GlcNAc2 + alpha-D-glucose
show the reaction diagram
Glc2Man9GlcNAc2 + H2O
Man9GlcNAc2 + D-glucose
show the reaction diagram
-
enzyme expression in Schizosaccharomyces pombe mutants either glucosidase II-alpha or both glucosidase II-alpha and -beta minus after cells transformed with the cDNA sequences of Arabidopsis thaliana glucosidase II-alpha encoding gene
-
?
Glc2Man9GlcNAc2 + H2O
Man9GlcNAc2 + glucose
show the reaction diagram
GlcMan7GlcNAc2 + H2O
Man7GlcNAc2 + D-glucose
show the reaction diagram
-
-
-
-
?
GlcMan9GlcNAc + H2O
Man9GlcNAc + D-glucose
show the reaction diagram
-
enzyme expression in Schizosaccharomyces pombe mutants either glucosidase II-alpha or both glucosidase II-alpha and -beta minus after cells transformed with the cDNA sequences of Arabidopsis thaliana glucosidase II-alpha encoding gene
-
?
GlcMan9GlcNAc2 + H2O
Man9GlcNAc2 + D-glucopyranose
show the reaction diagram
GlcMan9GlcNAc2 + H2O
Man9GlcNAc2 + D-glucose
show the reaction diagram
-
-
-
-
?
GlcNAcalpha(1->3)GlcNAcalpha(1->3)Manalpha(1->2)Manalpha(1->2)Manalpha(1->3)[Manalpha(1->2)Manalpha(1->6)[Manalpha(1->2)Manalpha(1->3)]Manalpha(1->6)]Manbeta(1->4)GlcNAcbeta(1->4)GlcNAc + H2O
GlcNAcalpha(1->3)Manalpha(1->2)Manalpha(1->2)Manalpha(1->3)[Manalpha(1->2)Manalpha(1->6)[Manalpha(1->2)Manalpha(1->3)]Manalpha(1->6)]Manbeta(1->4)GlcNAcbeta(1->4)GlcNAc + D-glucose
show the reaction diagram
GlcNAcalpha(1->3)GlcNAcalpha(1->3)Manalpha(1->2)Manalpha(1->2)Manalpha(1->3)[Manalpha(1->2)Manalpha(1->6)[Manalpha(1->2)Manalpha(1->3)]Manalpha(1->6)]Manbeta(1->4)GlcNAcbeta(1->4)GlcNAc-Gly-BODIPY + H2O
GlcNAcalpha(1->3)Manalpha(1->2)Manalpha(1->2)Manalpha(1->3)[Manalpha(1->2)Manalpha(1->6)[Manalpha(1->2)Manalpha(1->3)]Manalpha(1->6)]Manbeta(1->4)GlcNAcbeta(1->4)GlcNAc-Gly-BODIPY + D-glucose
show the reaction diagram
-
-
-
-
?
GlcNAcalpha(1->3)Manalpha(1->2)Manalpha(1->2)Manalpha(1->3)[Manalpha(1->2)Manalpha(1->6)[Manalpha(1->2)Manalpha(1->3)]Manalpha(1->6)]Manbeta(1->4)GlcNAcbeta(1->4)GlcNAc + H2O
Manalpha(1->2)Manalpha(1->2)Manalpha(1->3)[Manalpha(1->2)Manalpha(1->6)[Manalpha(1->2)Manalpha(1->3)]Manalpha(1->6)]Manbeta(1->4)GlcNAcbeta(1->4)GlcNAc + D-glucose
show the reaction diagram
GlcNAcalpha(1->3)Manalpha(1->2)Manalpha(1->2)Manalpha(1->3)[Manalpha(1->2)Manalpha(1->6)[Manalpha(1->2)Manalpha(1->3)]Manalpha(1->6)]Manbeta(1->4)GlcNAcbeta(1->4)GlcNAc-Gly-BODIPY + H2O
Manalpha(1->2)Manalpha(1->2)Manalpha(1->3)[Manalpha(1->2)Manalpha(1->6)[Manalpha(1->2)Manalpha(1->3)]Manalpha(1->6)]Manbeta(1->4)GlcNAcbeta(1->4)GlcNAc-Gly-BODIPY + D-glucose
show the reaction diagram
-
-
-
-
?
isomaltose + H2O
?
show the reaction diagram
kojibiose + H2O
?
show the reaction diagram
maltose + H2O
?
show the reaction diagram
maltose + H2O
alpha-D-glucose
show the reaction diagram
-
-
-
-
?
maltose + H2O
alpha-D-glucose + 1,5-anhydrofructose
show the reaction diagram
mannose oligosaccharide + H2O
?
show the reaction diagram
-
substrate specificity of wild-type and mutant enzymes with different mannose oligosaccharides, overview
-
-
?
mutan
glucan
show the reaction diagram
nigerose + H2O
?
show the reaction diagram
nigerose + H2O
alpha-D-glucose
show the reaction diagram
p-nitrophenyl-2-deoxy-alpha-D-glucopyranoside + H2O
p-nitrophenol + 2-deoxy-alpha-D-glucose
show the reaction diagram
-
very effective substrate, other deoxy derivatives are not hydrolyzed
-
-
?
pseudonigeran + H2O
alpha-D-glucose
show the reaction diagram
sucrose + H2O
D-glucose + D-fructose
show the reaction diagram
synthetic high-mannose-type glycan + H2O
?
show the reaction diagram
-
-
-
-
?
trehalose + H2O
?
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
alpha-1,3-mutan + H2O
alpha-D-glucose
show the reaction diagram
Q96VT3
involved in cell wall synthesis and sexual development
-
-
?
Glc2Man9GlcNAc2 (G2M9)-protein + H2O
?
show the reaction diagram
-
glucosidase II plays a key role in glycoprotein processing in the endoplasmic reticulum. This enzyme trims two alpha-1,3-linked glucose residues, Glcalpha1,3Glc (cleavage 1) and Glcalpha1,3Man (cleavage 2) from high-mannose type Glc2Man9GlcNAc2 (G2M9)-proteins. A crowded milieu that contains bovine serum albumin greatly enhances the second trimming step (cleavage 2), which deglucosylates Glc1Man9GlcNAc2, but not the first trimming step (cleavage 1), which removes the terminal glucose residue from Glc2Man9GlcNAc2
-
-
?
Glc2Man9GlcNAc2 + H2O
GlcMan9GlcNAc2 + D-glucopyranose
show the reaction diagram
Glc2Man9GlcNAc2 + H2O
Man9GlcNAc2 + alpha-D-glucose
show the reaction diagram
GlcMan9GlcNAc2 + H2O
Man9GlcNAc2 + D-glucopyranose
show the reaction diagram
mutan
glucan
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
-
activates slightly
Na+
-
activates slightly
additional information
-
no or poor effects by Ba2+, K+, and Fe2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-deoxynojirimycin
2,6-anhydro-1-benzamide-D-glycero-D-ido-heptitol
-
-
-
2,6-Dideoxy-2,6-imino-7-O-(beta-D-glucopyranosyl)-D-glycero-L-glucoheptitol
-
2-propanol
-
-
6-epicastanospermine
-
poor inhibitor
bromoconduritol
calreticulin
-
slower hydrolysis of Glc1Man9-residue to Glc0Man9-residue
-
castanospermine
Co2+
-
moderate inhibition at 1 mM
Cu2+
-
moderate inhibition at 1 mM
D-glucono-1,5-lactone
-
-
deoxynojirimycin
Epicatechin gallate
-
-
epigallocatechin
-
-
epigallocatechin gallate
-
-
gallocatechin
-
-
gallocatechin gallate
-
-
glycerol
-
-
isobutanol
-
-
maltose
Man7GlcNAc2
-
-
Man9GlcNAc2
-
-
methanol
-
-
Mn2+
-
moderate inhibition at 1 mM
N-5-Carboxypentyl-1-deoxynojirimycin
-
-
n-butanol
-
-
N-butyldeoxynojirimycin
-
-
n-Propanol
-
-
nigerose
p-chloromercuribenzenesulfonate
-
-
p-nitrophenyl-alpha-D-glucoside
-
-
p-nitrophenyl-alpha-D-mannoside
-
-
p-nitrophenyl-beta-D-glucoside
-
-
p-nitrophenyl-beta-D-mannoside
-
-
propyl gallate
-
-
Sn2+
-
moderate inhibition at 1 mM
turanose
validamine
-
-
valienamine
-
-
valiolamine
-
-
Zn2+
-
moderate inhibition at 1 mM
additional information
-
not inhibited by australine
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-deoxy-D-glucose
-
activation substrate p-nitrophenyl-alpha-D-glucoside
alpha-1,3-glucan
-
streptococcal mutan, induction
calreticulin
-
75 micromol, conversion of Glc2Man7-residue to Glc1Man7-residue; in higher concentrations faster hydrolysis of Glc2Man9-residue
-
cell wall material from fruiting bodies of Laetiporus sulphureus
-
induction
-
GlcMan9
-
faster reaction
isomaltose
-
lowering Km, 142% activation at 50 mM
mannose
-
increasing Vmax, 172% activation at 10 mM
starch
-
increasing Vmax, 160% activation at 7 mg/ml
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.013 - 0.019
4-methylumbelliferyl alpha-D-glucopyranoside
0.0003
4-Methylumbelliferyl-alpha-D-glucopyranoside
-
in 25 mM HEPES buffer, pH 7.2, at 37°C
0.0552
4-methylumbelliferyl-alpha-D-glucoside
-
37°C
2.2
4-nitrophenyl alpha-D-glucopyranoside
-
37°C
0.43 - 57.7
maltose
0.78 - 481
p-nitrophenyl alpha-D-glucopyranoside
0.76
p-nitrophenyl-2-deoxy-alpha-D-glucopyranoside
-
pH 6.8, 37°C
0.5 - 0.92
p-nitrophenyl-alpha-D-glucopyranoside
7.1 - 13
pseudonigeran
-
additional information
additional information
-
Km is 0.73 mg/ml with lyophilized Laetiporus sulphureus cell walls, pH 5.5, 45°C
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.015 - 0.017
castanospermine
0.003
deoxynojirimycin
pH 7.5, 37°C
0.008763 - 0.0131
Epicatechin gallate
0.06815 - 0.07599
epigallocatechin
0.02948 - 0.03281
epigallocatechin gallate
0.07062 - 0.07893
gallocatechin
0.002027 - 0.002545
gallocatechin gallate
0.002247 - 0.00519
N-butyldeoxynojirimycin
0.03628 - 0.04312
propyl gallate
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00024 - 0.00074
1-deoxynojirimycin
0.041 - 0.2616
bromoconduritol
0.0011 - 0.0068
castanospermine
0.00082 - 0.0112
deoxynojirimycin
4
Man7GlcNAc2
Rattus norvegicus
-
-
40
Man9GlcNAc2
Rattus norvegicus
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0535
-
soluble enzyme extract, using 4-methylumbelliferyl-alpha-D-glucopyranoside as substrate, in 25 mM HEPES buffer, pH 7.2, at 37°C
0.08
-
measured with substrate p-nitrophenyl-alpha-D-pyranosile
0.096
-
4-methylumbelliferyl alpha-D-glucopyranoside as substrate
0.151
-
4-methylumbelliferyl alpha-D-glucopyranoside as substrate
0.178
-
after incubation for 1 h at 45°C, medium supplementation with 2.0% cell wall preparation of Laetiporus sulphureus
0.442
-
p-nitrophenyl alpha-D-glucopyranoside as substrate
0.457
-
after 8.5fold purification, using 4-methylumbelliferyl-alpha-D-glucopyranoside as substrate, in 25 mM HEPES buffer, pH 7.2, at 37°C
0.501
-
-
0.617
-
p-nitrophenyl-alpha-D-glucopyranoside as substrate
0.782
-
active peptide fragment after trypsinization
1.72
-
after incubation for 1 h at 45°C, medium supplementation with 0.4% cell wall preparation of Laetiporus sulphureus
5.6
E16590
incubation at 35°C for 10 min, stop reaction by adding 0.4 ml of dinitrosalicylic acid solution, N-terminal domain with mutan-binding activity, C-terminal domain with mutanase activity and only low mutan-binding activity
115.6
-
purified enzyme with lyophilized Laetiporus sulphureus cell walls, pH 5.5, 45°C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 7.5
-
-
6.5 - 7
6.5
-
-
6.6 - 7
-
depending on substrate
6.7
-
-
6.8 - 7.5
-
-
7 - 8
-
30 mM maltose
7.4
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 7.5
-
citrate/phosphate buffer
4 - 7
E16590
80% of residual activity in comparison to the activity at ph 4.0
6.8 - 8.2
-
HEPES buffer
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.9
calculated from amino acid sequence
5.9
calculated from sequence
7.1
-
isoelectric focusing
7.5
-
basic chromatofocusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
T-lymphoma cell line
Manually annotated by BRENDA team
-
CD45-negative variant of BW5147
Manually annotated by BRENDA team
-
cultured T-lymphoma cells
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
; predominantly
Manually annotated by BRENDA team
additional information
-
subcellular localisation
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
11000
-
alpha, beta, 1 * 11000 + 1 * 80000, catalytic alpha subunit and beta subunit of unknown function
36000
-
gel filtration
47000
-
gel filtration
48000
calculated from amino acid sequence
58000
-
alphabeta 1 * 104000 + 1 * 58000
63800
-
calculated from amino acid sequence
73000
12 * 73000, calculated from sequence
75400
-
2 * 75400, SDS-PAGE
84000
-
SDS-PAGE, beta subunit with unknown function
100000 - 123000
100000
-
4 * 100000, SDS-PAGE
104000
106000
-
alpha4, 4 * 106000
107000
-
may be a proteolytic degradation product of the 112000 protein
110000
112000
-
-
116000
117000
molecular mass of protein extract of both glucosidase II-alpha and -beta minus Schizosaccharomyces pombe mutant cells after cells transformed with the cDNA sequences of Arabidopsis thaliana glucosidase II-alpha encoding gen and after deglycosylation
119000
western blot analysis of protein extract of both glucosidase II-alpha and -beta minus Schizosaccharomyces pombe mutant cells after cells transformed with the cDNA sequences of Arabidopsis thaliana glucosidase II-alpha encoding gene
135000
E16590
by MALDI-TOF mass spectrometry
152700
-
gel filtration
161000
-
analytical ultracentrifugation
220000
-
gel filtration
250000
native PAGE
260000 - 290000
400000
-
non denaturing PAGE
425000
-
gel filtration
914000
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dodecamer
heterodimer
homodimer
monomer
tetramer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
-
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5 - 7
-
at 4°C
136906
3.5 - 6
-
at 30°C
136907
3.5 - 7
-
at 4°C
136907
4 - 7
-
at 30°C
136906
4.5 - 6
-
purified enzyme, 45°C, 1 h, stable at
731044
6 - 8
-
with 5 mM mercaptoethanol
136904
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
non-permissive stability
38
-
at pH 7.0
43
-
up to at pH 3.5
50
-
up to
60
E16590
above unstable, stable for 10 min
95
half-life: 33.8 h
100
half-life: 10.6 h
105
half-life: 1.8 h
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
alpha-1,3-glucan stabilizes the enzyme
-
glycerol, stabilization
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 100 mM acetate buffer, pH 5.5
-
-20°C, purified enzyme, 4 days, 50% loss of activity
-
-70°C, 0.9% NaCl, several months
-
-70°C, 100 mM phosphate buffer, pH 7, 100 mM maltose, 5 mM mercaptoethanol, several weeks
-
4-6°C, 100 mM phosphate buffer, pH 7, 10% glycerol, 5 mM mercaptoethanol, several days
-
4°C, 10 mM potassium phosphate buffer, pH 7, 0.1% Triton X-100, 5 mM 2-mercaptoethanol, 14 days
-
4°C, 10 mM potassium phosphate buffer, pH 7, 0.1% Triton X-100, 5 mM 2-mercaptoethanol, 8 days ethanol treated rats
-
4°C, purified enzyme, 2 days, 50% loss of activity
-
4°C, soluble extract, 6 days, 50% loss of activity
-
4°C, stored for at least 6 months without any loss of activity
below 60°C, 10 min
E16590
freezing inactivates the enzyme
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2-fold purification and 98% residual enzyme activity after precipitation with propanol, around 2-fold purification and 77% enzyme recovery after salting out with ammonium sulfate at 50% saturation, 10-fold concentrated preparation of the enzyme with a yield of 98% after ultrafiltration, mutanase recovery of 97% after lyophilization and concentration of the culture broth in a vacuum evaporator
-
by chromatographic procedures
E16590
Mono Q column chromatography and Sephacryl S-300 gel filtration
-
native extracellular enzyme 18.4fold by ultrafiltration, anion exchange and hydrophobic interaction chromatography, and chromatofocusing to homogeneity
-
native protein and alpha subunit
-
purification of recombinant protein using His-tag
recombinant proteins from Escherichia coli
-
yield of 56.3% of alkali-soluble and water-insoluble D-glucan
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning and characterization of the glucosidase II alpha subunit gene
-
cloning of alpha- and beta-subunits and expression in Escherichia coli strains DH5alpha and JA226
-
construction of a cDNA library
expressed in CHO cells
-
expressed in Sulfolobus acidocaldarius. Attempts to obtain soluble enzyme from Escherichia coli strains are unsuccessful
expressed with a baculovirus expression system in Sf9 cells, expressed in Cos7 cells
expression in Cos-1 cells
expression in Escherichia coli
-
expression in Schizosaccharomyces pombe mutants either glucosidase II-alpha or both glucosidase II-alpha and -beta minus, vector pTOTO, vector pREP3X, vector pSGP72
expression of wild-type and mutant beta-subunits and enzymes in HeLaS3 cells, overexpression in HEK 293T cells
-
expression of wild-type and mutant fusion forms of the enzyme in Sf9 insect cells
fragments and full length beta-subunit expressed as GST-fusion protein in Escherichia coli
-
genes Aogls2alpha and/or Aogls2beta encdoing the alpha- und beta-subunit, cloning and expression in Escherichia coli strain DH5alpha
-
recombinant proteins in E. coli strains NV522 an B221
E16590
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
ethanol-sensitive GIIbeta is upregulated by about 70% in ethanol-exposed cerebral cortex
-
in live Schizosaccharomyces pombe cells, a decrease in the number of mannoses in the glycan results in decreased glucosidase II activity
the transcription level of malA is increased 3fold upon the addition of maltose or starch to the medium
the water-insoluble fraction of Laetiporus sulphureus fruiting bodies from 0.15 to 0.2% (w/v) induces mutanase activity in Paenibacillus sp. strain MP-1
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
rsw3
Substitution of Ser599 by a Phe residue, inactivation of the alpha-subunit under non-permissive temperature because mutant is strongly temperature-sensitive
D542N
D542 required for catalytic activity
D564N
D564 required for catalytic activity
Q420E
-
site-directed mutagenesis of the mannose 6-phosphate receptor homology domain of the beta-subunit, GIIbeta-MRH, leading to reduced activity with substrates G1M9 and G2M9
Y410A
-
site-directed mutagenesis of the mannose 6-phosphate receptor homology domain of the beta-subunit, GIIbeta-MRH, leading to reduced activity with substrates G1M9 and G2M9
E114A
-
inactive towards Glc1Man9GlcNAc2
E73A
-
inactive towards Glc1Man9GlcNAc2
D564E
42.7% of wild-type activity
D564N
no activity detectable
E567D
53.2% of wild-type activity
E567Q
no activity detectable
F571A
74.8% of wild-type activity
E114A
-
inactive towards Glc1Man9GlcNAc2
E457Q
-
site-directed mutagenesis
E73A
-
inactive towards Glc1Man9GlcNAc2
Q408E
-
site-directed mutagenesis
R438K
-
site-directed mutagenesis
Y463F
-
site-directed mutagenesis
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
medicine