Information on EC 3.2.1.70 - glucan 1,6-alpha-glucosidase

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

EC NUMBER
COMMENTARY hide
3.2.1.70
-
RECOMMENDED NAME
GeneOntology No.
glucan 1,6-alpha-glucosidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of (1->6)-alpha-D-glucosidic linkages in (1->6)-alpha-D-glucans and derived oligosaccharides
show the reaction diagram
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-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
non-pathway related
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SYSTEMATIC NAME
IUBMB Comments
glucan 6-alpha-D-glucohydrolase
Hydrolysis is accompanied by inversion at C-1, so that new reducing ends are released in the beta-configuration. Dextrans and isomaltosaccharides are hydrolysed, as is isomaltose, but very slowly. The enzyme from some sources also possesses the activity of EC 3.2.1.59 (glucan endo-1,3-alpha-glucosidase).
CAS REGISTRY NUMBER
COMMENTARY hide
37288-48-5
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
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Manually annotated by BRENDA team
Bacillus sp. LMM 12
LMM 12
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Manually annotated by BRENDA team
Bacillus sp. SAM 1606
SAM 1606
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Manually annotated by BRENDA team
gene LBA0264
UniProt
Manually annotated by BRENDA team
gene LBA0264
UniProt
Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
gene dexB; serotype C, gene dexB
Uniprot
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
physiological function
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
4-nitrophenyl alpha-D-glucopyranoside + H2O
4-nitrophenol + alpha-D-glucopyranose
show the reaction diagram
-
-
-
-
?
4-nitrophenyl alpha-D-glucoside + H2O
4-nitrophenol + D-glucose
show the reaction diagram
377% of the activity with isomaltose, wild-type enzyme
-
-
?
4-nitrophenyl-alpha-D-glucopyranoside + H2O
4-nitrophenol + D-glucose
show the reaction diagram
4-nitrophenyl-alpha-isomaltoside + H2O
4-nitrophenol + isomaltose
show the reaction diagram
dextran + H2O
?
show the reaction diagram
dextran + H2O
alpha-D-glucose
show the reaction diagram
-
-
-
?
dextran + H2O
beta-D-glucose
show the reaction diagram
dextran + H2O
beta-D-glucose + ?
show the reaction diagram
-
-
-
-
?
dextran + H2O
D-glucose
show the reaction diagram
-
-
-
-
?
dextran + H2O
D-glucose + ?
show the reaction diagram
dextran + H2O
glucose
show the reaction diagram
dextrans + H2O
beta-D-glucose
show the reaction diagram
glycogen + H2O
?
show the reaction diagram
-
-
-
-
?
isomaltodextrins + H2O
beta-D-glucose
show the reaction diagram
isomaltoheptaose + H2O
D-glucose
show the reaction diagram
148% of the activity with isomaltose, wild-type enzyme
-
-
?
isomaltohexaose + H2O
D-glucose
show the reaction diagram
148% of the activity with isomaltose, wild-type enzyme
-
-
?
isomaltooligosaccharide + H2O
D-glucose + ?
show the reaction diagram
isomaltooligosacchrides + H2O
D-glucose
show the reaction diagram
-
-
-
-
?
isomaltopentaose + H2O
beta-D-glucose
show the reaction diagram
-
-
-
?
isomaltopentaose + H2O
D-glucose
show the reaction diagram
148% of the activity with isomaltose, wild-type enzyme
-
-
?
isomaltose + H2O
2 D-glucose
show the reaction diagram
isomaltose + H2O
beta-D-glucose
show the reaction diagram
isomaltose + H2O
D-glucose
show the reaction diagram
-
-
-
?
isomaltotetraose + H2O
?
show the reaction diagram
isomaltotetraose + H2O
beta-D-glucose
show the reaction diagram
-
-
-
?
isomaltotetraose + H2O
D-glucose
show the reaction diagram
148% of the activity with isomaltose, wild-type enzyme
-
-
?
isomaltotriose + H2O
?
show the reaction diagram
isomaltotriose + H2O
beta-D-glucose
show the reaction diagram
isomaltotriose + H2O
D-glucose
show the reaction diagram
kojibiose + H2O
D-glucose
show the reaction diagram
4.02% of the activity with isomaltose, wild-type enzyme
-
-
?
maltose + H2O
D-glucose
show the reaction diagram
0.0135% of the activity with isomaltose, wild-type enzyme
-
-
?
maltotetraose + H2O
D-glucose
show the reaction diagram
0.0108% of the activity with isomaltose, wild-type enzyme
-
-
?
maltotriose + H2O
D-glucose
show the reaction diagram
0.146% of the activity with isomaltose, wild-type enzyme
-
-
?
nigerose + H2O
2 D-glucose
show the reaction diagram
1.99% of the activity with isomaltose, wild-type enzyme
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-
?
p-nitrophenyl-alpha-D-glucopyranoside + H2O
p-nitrophenol + alpha-D-glucopyranose
show the reaction diagram
panose + H2O
beta-D-glucose
show the reaction diagram
-
-
-
?
panose + H2O
D-glucose
show the reaction diagram
panose + H2O
D-glucose + ?
show the reaction diagram
sucrose + H2O
D-fructose + D-glucose
show the reaction diagram
4.29% of the activity with isomaltose, wild-type enzyme
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?
additional information
?
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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
dextran + H2O
beta-D-glucose + ?
show the reaction diagram
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-
-
-
?
dextran + H2O
D-glucose + ?
show the reaction diagram
dextrans + H2O
beta-D-glucose
show the reaction diagram
isomaltooligosaccharide + H2O
D-glucose + ?
show the reaction diagram
Q5FMB7
degradation
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-
?
isomaltose + H2O
2 D-glucose
show the reaction diagram
isomaltotetraose + H2O
?
show the reaction diagram
isomaltotriose + H2O
?
show the reaction diagram
panose + H2O
D-glucose + ?
show the reaction diagram
Q5FMB7
-
-
-
?
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
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the addition of Mg2+ (optimum 15 mM) enhances the thermostability of dextran glucosidase
additional information
-
K+ and Na+ do not enhance the thermostability of dextran glucosidase
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
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structural lid (acids 99-97) at the active site generated by the tetramerization is closely associated with the bifunctionality and in particular with the alpha-1,4-transferase activity
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.38
4-nitrophenyl-alpha-D-glucopyranoside
purified recombinant enzyme, pH 6.0, 37C
0.012 - 29.1
Dextran
12 - 81.7
isomaltoheptaose
7.88 - 76.9
isomaltohexaose
5.1 - 38.3
isomaltopentaose
8.91 - 22.5
isomaltose
5.58 - 20.9
isomaltotetraose
3.17 - 14.1
isomaltotriose
0.549 - 41.4
p-nitrophenyl alpha-D-glucoside
1.03 - 6.41
panose
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
597
4-nitrophenyl-alpha-D-glucopyranoside
Lactobacillus acidophilus
Q5FMB7
purified recombinant enzyme, pH 6.0, 37C
95.2 - 384
Dextran
9.81 - 449
isomaltoheptaose
3 - 401
isomaltohexaose
0.04 - 419
isomaltopentaose
8.88 - 517
isomaltose
0.04 - 501
isomaltotetraose
12.2 - 541
isomaltotriose
61.4 - 1202
p-nitrophenyl alpha-D-glucoside
22.4 - 612
panose
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
251
4-nitrophenyl-alpha-D-glucopyranoside
Lactobacillus acidophilus
Q5FMB7
purified recombinant enzyme, pH 6.0, 37C
4620
13.6
Dextran
Lactobacillus acidophilus
Q5FMB7
purified recombinant enzyme, pH 6.0, 37C
1045
23
isomaltose
Lactobacillus acidophilus
Q5FMB7
purified recombinant enzyme, pH 6.0, 37C
449
31
isomaltotetraose
Lactobacillus acidophilus
Q5FMB7
purified recombinant enzyme, pH 6.0, 37C
2848
34
isomaltotriose
Lactobacillus acidophilus
Q5FMB7
purified recombinant enzyme, pH 6.0, 37C
1469
157
panose
Lactobacillus acidophilus
Q5FMB7
purified recombinant enzyme, pH 6.0, 37C
1065
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
40
D-glucose
pH 6.0, 37C, recombinant enzyme
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
34.3 - 38.4
335
purified recombinant enzyme, pH 6.0, 37C
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 7.4
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more than 80% of maximal activity betwenn pH 5.5 and pH 7.4
6 - 7
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no activity below pH 5.0 or above pH 9.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
assay at; assay at
50
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50% of activity at 40C, 10% of activity at 60C
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
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50% of activity at 40C, 10% of activity at 60C
60
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not active above
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
recombinant His-tagged enzyme, isoelectric focusing
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Streptococcus mutans serotype c (strain ATCC 700610 / UA159)
Streptococcus mutans serotype c (strain ATCC 700610 / UA159)
Streptococcus mutans serotype c (strain ATCC 700610 / UA159)
Streptococcus mutans serotype c (strain ATCC 700610 / UA159)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
29000
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gel filtration
63000
wild-type enzyme, gel filtration
70000
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gel filtration
109135
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x * 109135, calculated from sequence
120000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
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crystallography
monomer
tetramer
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crystallography
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
determination of the structure of glucodextranase and of the complex of glucodextranase with acarbose at 2.42 A resolution
unliganded form and in complex with acarbose
purified recombinant His-tagged enzyme, sitting drop vapour diffusion method, mixing of 16 mg/ml protein in 20 mM MES-NaOH, pH 6.5, 100 mM NaCl, and 2 mM CaCl2, with reservoir solution containing 20% glycerol, 16% PEG 8000, and 0.1 M MES, pH 6.5, in a 2:1 ratio, room temperature, X-ray diffraction structure determination and analysis at 2.05 A resolution, molecular replacement and modelling
by hanging drop vapour-diffusion method, using polyethylene glycol 6000 as precipitant
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crystal structures of DGase in uncomplexed form and a mutant in complex with isomaltotriose, to 2.2 A resolution, by the hanging-drop, vapor-diffusion method at 20C. The enzyme is composed of three domains, A, B and C, and has a (beta/alpha)8-barrel in domain A. Three catalytic residues are located at the bottom of the active site pocket and the bound isomaltotriose occupies subsites -1 to +2. Hydrogen bonds between Asp60 and Arg398 and O4 atom of the glucose unit at subsite -1 accomplish recognition of the non-reducing end of the bound substrate. The side-chain atoms of Glu371 and Lys275 form hydrogen bonds with the O2 and O3 atoms of the glucose residue at subsite +1
at a resolution of 2.8 A, TreX crystallized in the dimeric and tetrameric forms, in space groups P3121 and P321, respectively and in complex with an acarbose ligand. Acarbose intermediate is covalently bound to Asp363, occupying subsites -1 to -3
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 8
purified recombinant His6-tagged enzyme, over 50% activity remaining
732004
4.5 - 6
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208769
5.5 - 7.5
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208767
5.5 - 7
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the enzyme is stable between pH 5.5 and 7.0 in the presence of 5 mM Ca2+ after 15 min at 37C
714533
5.5 - 8.4
4C, stable for 18 h
664233
5.8 - 6.7
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the enzyme is stable between pH 5.8 and 6.7 in the absence of Ca2+ after 15 min at 37C
714533
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
purified recombinant His6-tagged enzyme, pH 6.0, inactivation rate constant is 0.0001/min, T1/2 at 37C is 4.8 days
40
15 min, pH 6.0, stable below
46
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the enzyme activity drops to 50% in 5 min at 46C at 0-1 mM Ca2+, but the presence of 2 mM Ca2+ prolongs this to 30 min and 3-30 mM Ca2+ stabilizes the enzyme so that it retains 70% activity or higher even after 60 min incubation at 46C
60
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pH 7.2, stable up to, wild type and several deletion mutants
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by Ni-chelating column chromatography
by nickel-chelating chromatography with Chelating Sepharose FF
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recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, anion exchange chromatography, and ultrafiltration, to homogeneity
wild-type enzyme and mutant enzymes E458Q and E656Q
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
dexB, sequence comparisons; dexB, sequence comparisons, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) CodonPlus RIL; dexB, sequence comparisons, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) CodonPlus RIL
expressed in Escherichia coli BL21(DE3) CodonPlus RIL cells
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expression in Escherichia coli
gene LBA0264, DNA and amino acid sequence determination and analysis, genetic organization, sequence comparisons and phylogenetic tree, the enzyme is encoded in the maltooligosaccharide operon, semiquantitative PCR expression analysis, cloning in Escherichia coli strain TOP10, expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
wild-type and mutant enzymes produced in Escherichia coli
wild-type and mutant expressed in Escherichia coli BL21 (DE3) CodonPlus RIL containing the expression plasmid derived from pET23d
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
besides the isolated locus encoding a GH13_31 enzyme, the ABC transporter and another GH13 in the maltooligosaccharide operon are induced in response to IMO or maltotetraose suggesting coregulation of alpha-1,6- and alpha-1,4-glucooligosaccharide utilization loci in Lactobacillus acidophilus NCFM
the enzyme is induced by isomaltooligosaccharides and maltotetraose
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E458Q
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mutant enzyme shows 0.0005% of the wild-type activity
E656Q
-
mutant enzyme shows 0.0003% of the wild-type activity
E458Q
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mutant enzyme shows 0.0005% of the wild-type activity
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E656Q
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mutant enzyme shows 0.0003% of the wild-type activity
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C129S/C532S
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme, the mutant predominantly catalyzes hydrolysis; site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme, the mutant predominantly catalyzes hydrolysis
D194A/C129S/C532S
site-directed mutagenesis; site-directed mutagenesis
D194C/C129S/C532S
site-directed mutagenesis, the mutant shows highly reduced activity compared with the wild-type enzyme and mutant C129S/C532S, oxidation with KI activates the mutant by 330fold which is 0.27% activity of the activity of mutant C129S/C532S, the oxidized mutant Ox-D194C-2CS catalyzes both hydrolysis and transglucosylation; site-directed mutagenesis, the mutant shows highly reduced activity compared with the wild-type enzyme and mutant C129S/C532S, oxidation with KI activates the mutant by 330fold which is 0.27% activity of the activity of mutant C129S/C532S, the oxidized mutant Ox-D194C-2CS catalyzes both hydrolysis and transglucosylation
E236Q
catalysis is compromised in complex with isomaltotriose
W238A
mutant shows lower preference for kojibiose, nigerose, and sucrose and higher preference for maltooligosaccharides, trehalose, and p-nitrophenyl alpha-D-glucoside. Altough trehalose is not a substrate for the wild-type, the mutant enzyme hydrolyzes trehalose
W238N
mutant shows lower preference for kojibiose, nigerose, and sucrose and higher preference for maltooligosaccharides, trehalose, and p-nitrophenyl alpha-D-glucoside. Altough trehalose is not a substrate for the wild-type, the mutant enzyme hydrolyzes trehalose
W238P
mutant shows lower preference for kojibiose, nigerose, and sucrose and higher preference for maltooligosaccharides, trehalose, and p-nitrophenyl alpha-D-glucoside. Altough trehalose is not a substrate for the wild-type, the mutant enzyme hydrolyzes trehalose
C129S/C532S
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site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme, the mutant predominantly catalyzes hydrolysis
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D194A/C129S/C532S
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site-directed mutagenesis
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D194C/C129S/C532S
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site-directed mutagenesis, the mutant shows highly reduced activity compared with the wild-type enzyme and mutant C129S/C532S, oxidation with KI activates the mutant by 330fold which is 0.27% activity of the activity of mutant C129S/C532S, the oxidized mutant Ox-D194C-2CS catalyzes both hydrolysis and transglucosylation
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D318A
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shows an activity level identical to that of the wild-type
E94A
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shows a relatively minor change in the alpha-1,6-glucosidase activity, but a sharply increased alpha-1,4-transferase activity compared with the wild-type
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
medicine
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the use of iminosugars is applied in preventing dental caries by inhibiting the activity of the dextran glucosidases
additional information
-
bifunctional mechanism of the archaeal glycogen-debranching enzyme TreX, showing alpha-1,4-transferase and alpha-1,6-glucosidase activities
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