Information on EC 2.4.1.20 - cellobiose phosphorylase

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The expected taxonomic range for this enzyme is: Bacteria

EC NUMBER
COMMENTARY hide
2.4.1.20
-
RECOMMENDED NAME
GeneOntology No.
cellobiose phosphorylase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
cellobiose + phosphate = alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexosyl group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Starch and sucrose metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
cellobiose:phosphate alpha-D-glucosyltransferase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9030-20-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
Cellvibrio gilvus
Clostridium stercorarium
strain NCIB 11754
SwissProt
Manually annotated by BRENDA team
Clostridium stercorarium NCIB 11754
strain NCIB 11754
SwissProt
Manually annotated by BRENDA team
Fomes annosus
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain MSB8
-
-
Manually annotated by BRENDA team
-
O52504
SwissProt
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,5-anhydroglucitol + glucose-1-phosphate
?
show the reaction diagram
Cellvibrio gilvus
-
-
-
-
?
alpha-D -glucose 1-fluoride + D-glucose
cellobiose + fluoride
show the reaction diagram
-
-
-
-
ir
alpha-D-glucose 1-phosphate + 1,5-anhydro-D-glucitol
? + phosphate
show the reaction diagram
alpha-D-glucose 1-phosphate + 2-deoxy-2-fluoro-D-glucose
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + 2-deoxy-D-glucose
?
show the reaction diagram
alpha-D-glucose 1-phosphate + 2-deoxy-D-glucose
? + phosphate
show the reaction diagram
alpha-D-glucose 1-phosphate + 5a-carba-alpha-D,L-glucopyranose
4-O-beta-D-glucopyranosyl-5a-carba-beta-D-glucopyranose + phosphate
show the reaction diagram
Cellvibrio gilvus
-
very poor substrate
-
-
?
alpha-D-glucose 1-phosphate + 5a-carba-beta-D,L-glucopyranose
?
show the reaction diagram
Cellvibrio gilvus
-
much better substrate than alpha-anomer, 22% of activity with D-glucose
-
-
?
alpha-D-glucose 1-phosphate + 6-deoxy-6-fluoro-D-glucose
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + 6-deoxy-D-glucose
?
show the reaction diagram
alpha-D-glucose 1-phosphate + 6-deoxy-D-glucose
? + phosphate
show the reaction diagram
alpha-D-glucose 1-phosphate + butyl beta-D-glucoside
? + phosphate
show the reaction diagram
-
no substrate for wild-type
-
-
r
alpha-D-glucose 1-phosphate + cellobiose
? + phosphate
show the reaction diagram
-
no substrate for wilalpha-D-type
-
-
r
alpha-D-glucose 1-phosphate + D-altrose
?
show the reaction diagram
alpha-D-glucose 1-phosphate + D-arabinose
?
show the reaction diagram
alpha-D-glucose 1-phosphate + D-gentobiose
? + phosphate
show the reaction diagram
-
-
-
-
r
alpha-D-glucose 1-phosphate + D-glucosamine
?
show the reaction diagram
alpha-D-glucose 1-phosphate + D-glucosamine
? + phosphate
show the reaction diagram
-
-
-
-
r
alpha-D-glucose 1-phosphate + D-glucose
? + phosphate
show the reaction diagram
-
-
-
-
r
alpha-D-glucose 1-phosphate + D-glucose
cellobiose + phosphate
show the reaction diagram
alpha-D-glucose 1-phosphate + D-mannose
?
show the reaction diagram
alpha-D-glucose 1-phosphate + D-mannose
? + phosphate
show the reaction diagram
-
-
-
-
r
alpha-D-glucose 1-phosphate + D-xylose
?
show the reaction diagram
alpha-D-glucose 1-phosphate + D-xylose
? + phosphate
show the reaction diagram
-
-
-
-
r
alpha-D-glucose 1-phosphate + ethyl beta-D-glucoside
? + phosphate
show the reaction diagram
-
no substrate for wild-type
-
-
r
alpha-D-glucose 1-phosphate + L-fucose
?
show the reaction diagram
alpha-D-glucose 1-phosphate + L-galactose
?
show the reaction diagram
alpha-D-glucose 1-phosphate + methyl alpha-D-glucoside
? + phosphate
show the reaction diagram
-
no substrate for wild-type
-
-
r
alpha-D-glucose 1-phosphate + methyl beta-D-glucoside
? + phosphate
show the reaction diagram
-
no substrate for wild-type
-
-
r
alpha-D-glucose 1-phosphate + phenyl beta-D-glucoside
? + phosphate
show the reaction diagram
-
no substrate for wild-type
-
-
r
beta-D-glucose 1-phosphate + alpha-D-glucopyranosyl fluoride
beta-cellobiose + HF
show the reaction diagram
-
beta-D-glucopyranosyl fluoride is not ulilized
-
?
cellobiose + arsenate
D-glucose + alpha-D-glucose 1-arsenate
show the reaction diagram
-
-
alpha-D-glucose 1-arsenate hydrolyses spontaneously to form D-glucose and arsenate
ir
cellobiose + phosphate
alpha-D-glucose 1 phosphate + D-glucose
show the reaction diagram
-
-
-
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
cellobiose + phosphate
D-glucose 1-phosphate + D-glucose
show the reaction diagram
-
-
-
-
?
D-glucal + 2-deoxy-2-(hydroxymethyl)-beta-D-xylopyranose
2-deoxy-3-O-(2-deoxy-beta-D-arabino-hexopyranosyl)-2-(hydroxymethyl)-beta-D-xylopyranose + ?
show the reaction diagram
Cellvibrio gilvus
-
-
-
-
r
D-glucal + D-glucose
2-deoxy-beta-D-arabino-hexopyranosyl-(1->4)-D-glucose + ?
show the reaction diagram
Cellvibrio gilvus
-
-
-
-
r
D-glucal + D-mannose
2-deoxy-beta-D-arabino-hexopyranosyl-(1->4)-D-mannose + ?
show the reaction diagram
Cellvibrio gilvus
-
-
-
-
r
D-glucal + D-xylose
2-deoxy-beta-D-arabino-hexopyranosyl-(1->4)-D-xylose + ?
show the reaction diagram
Cellvibrio gilvus
-
-
-
-
r
D-glucal + phosphate
2-deoxy-alpha-D-glucose 1-phosphate
show the reaction diagram
-
D-glucal is a slow alternative donor substrate for stereospecific glycosyl transfer to phosphate. Reaction follows a stereochemical course where substrate becomes protonated from below its six-membered ring through stereoselective re side attack at C-2. The proposed catalytic mechanism involves direct protonation of D-glucal by the enzyme-bound phosphate, which then performs nucleophilic attack on the reactive C-1 of donor substrate. When offered D-glucose next to D-glucal and phosphate, the enzyme produces 2-deoxy-beta-D-glucosyl-(1->4)-D-glucose and 2-deoxy-alpha-D-glucose 1-phosphate in a ratio governed by mass action of the two acceptor substrates present. Enzymatic synthesis of 2-deoxy-beta-D-glucosyl-(1->4)-D-glucose is effectively promoted by catalytic concentrations of phosphate, suggesting that catalytic reaction proceeds through a quaternary complex of enzyme, D-glucal, phosphate,and D-glucose
-
-
?
gentiobiose + alpha-D-glucose 1-phosphate
?
show the reaction diagram
Cellvibrio gilvus
-
-
-
-
?
isomaltose + alpha-D-glucose 1-phosphate
?
show the reaction diagram
Cellvibrio gilvus
-
-
-
-
?
lactose + phosphate
alpha-D-galactose 1-phosphate + D-glucose
show the reaction diagram
-
-
-
r
melibiose + alpha-D-glucose 1-phosphate
?
show the reaction diagram
Cellvibrio gilvus
-
-
-
-
?
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-D-glucose 1-phosphate + D-glucose
cellobiose + phosphate
show the reaction diagram
Cellvibrio gilvus
-
-
-
-
r
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
no influence of metal ions on reaction
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-deoxy-D-glucose
-
competitive with D-glucose, dead end inhibitor
4-thio-cellobiose
-
weak inhibition in both directions of reaction
6-deoxy-D-glucose
Cellvibrio gilvus
-
substrate inhibition
alpha-oxogluconate
Cellvibrio gilvus
-
-
cellobiose
D-cellobiose
Cellvibrio gilvus
-
-
D-glucosamine
Cellvibrio gilvus
-
substrate inhibition
D-glucose
D-glucose 1-phosphate
glucono-delta-lactone
Cellvibrio gilvus
-
very weak
N-ethylmaleimide
Nojirimycin
p-chloromercuribenzoate
Cellvibrio gilvus
-
complete inhibition at 0.02 mM
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
9.5
1,5-Anhydro-D-glucitol
-
pH 6.0, 37C
8 - 9
1,5-anhydroglucitol
Cellvibrio gilvus
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
17.2
2-deoxy-2-fluoro-D-glucose
-
-
24.3 - 250
2-deoxy-D-glucose
7.4
6-deoxy-6-fluoro-D-glucose
-
-
4.1 - 24
6-deoxy-D-glucose
2.15 - 8
alpha-D-glucose 1-phosphate
33.7
butyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
0.29 - 16.5
cellobiose
2.6
D-cellobiose
Cellvibrio gilvus
-
-
10.8
D-gentobiose
-
pH 6.0, 37C
5.7 - 13.3
D-glucosamine
0.69 - 28.9
D-glucose
10.8
D-glucoseamine
-
-
-
23.5 - 170
D-mannose
14 - 84
D-xylose
138 - 155
ethyl beta-D-glucoside
83
gentiobiose
Cellvibrio gilvus
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
71
isomaltose
Cellvibrio gilvus
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
228 - 330
lactose
93
melibiose
Cellvibrio gilvus
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
1009
methyl alpha-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
56.6 - 79.8
methyl beta-D-glucoside
31.2
phenyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
0.15 - 2.9
phosphate
additional information
additional information
Cellvibrio gilvus
-
simulation of molecular dynamics of the enzyme, the 1S3 pre-transition state conformer is highly stable compared with other conformers, and a conformational change from 4C1 to 1,4B occurs
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.8
1,5-Anhydro-D-glucitol
Ruminococcus albus
-
pH 6.0, 37C
3.5
1,5-anhydroglucitol
Cellvibrio gilvus
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
8
2-deoxy-2-fluoro-D-glucose
Cellulomonas uda
-
-
11.6 - 21.5
2-deoxy-D-glucose
44.6
6-deoxy-6-fluoro-D-glucose
Cellulomonas uda
-
-
17 - 124
6-deoxy-D-glucose
43.2
alpha-D-glucose 1-phosphate
Cellulomonas uda
-
pH 6.6, 30C, native enzyme; pH 6.6, 30C, recombinant His6-tagged enzyme
9
butyl beta-D-glucoside
Cellulomonas uda
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
1.1 - 29.1
cellobiose
4
D-gentobiose
Ruminococcus albus
-
pH 6.0, 37C
5.2 - 9.56
D-glucosamine
5.3 - 92.3
D-glucose
5.6
D-glucoseamine
Cellulomonas uda
-
-
-
2.83 - 9.3
D-mannose
8.7 - 44.3
D-xylose
5.1 - 8.7
ethyl beta-D-glucoside
7.2
gentiobiose
Cellvibrio gilvus
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
1.9
isomaltose
Cellvibrio gilvus
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
0.109 - 1.11
lactose
2.3
melibiose
Cellvibrio gilvus
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
6
methyl alpha-D-glucoside
Cellulomonas uda
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
6.8 - 7.9
methyl beta-D-glucoside
4
phenyl beta-D-glucoside
Cellulomonas uda
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
15.4
phosphate
Cellulomonas uda
-
pH 6.6, 30C, native enzyme; pH 6.6, 30C, recombinant His6-tagged enzyme
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.29
1,5-Anhydro-D-glucitol
Ruminococcus albus
-
pH 6.0, 37C
2926
0.36
2-deoxy-D-glucose
Ruminococcus albus
-
pH 6.0, 37C
531
12.7
6-deoxy-D-glucose
Ruminococcus albus
-
pH 6.0, 37C
1527
0.266
butyl beta-D-glucoside
Cellulomonas uda
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
20171
0.103 - 4.56
cellobiose
0.37
D-gentobiose
Ruminococcus albus
-
pH 6.0, 37C
42200
0.72
D-glucosamine
Ruminococcus albus
-
pH 6.0, 37C
699
0.031 - 63.2
D-glucose
0.12
D-mannose
Ruminococcus albus
-
pH 6.0, 37C
216
0.65
D-xylose
Ruminococcus albus
-
pH 6.0, 37C
115
0.033 - 0.063
ethyl beta-D-glucoside
0.00048 - 0.0034
lactose
0.006
methyl alpha-D-glucoside
Cellulomonas uda
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
5684
0.099 - 0.12
methyl beta-D-glucoside
0.129
phenyl beta-D-glucoside
Cellulomonas uda
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
4858
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4.4
4-deoxy-D-glucose
-
-
13
4-thio-cellobiose
-
in presence of saturating concentrations of phosphate
14.8
cellobiose
-
-
6.8
D-cellobiose
Cellvibrio gilvus
-
-
1.2 - 255
D-glucose
3.2
D-glucose 1-phosphate
-
-
0.045
Nojirimycin
Cellvibrio gilvus
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.033
purifed recombinant wild-type enzyme, substrate lactose
0.1
purifed recombinant mutant A397V/T508A/A512T/D557N/N667T/G681S, substrate lactose
0.171
purifed recombinant mutant T508I/N667A, substrate lactose
0.249
purifed recombinant mutant T508A/N667T, substrate lactose
1.35
purifed recombinant mutant A397V/T508A/A512T/D557N/N667T/G681S, substrate cellobiose
1.71
purifed recombinant mutant T508I/N667A, substrate cellobiose
2.61
purifed recombinant mutant T508A/N667T, substrate cellobiose
2.66
-
purified enzyme
4.7
Clostridium stercorarium
purified enzyme
7.78
purifed recombinant wild-type enzyme, substrate cellobiose
14.4
-
pH 6.6, 30C
20
-
purified enzyme
27.4
Cellvibrio gilvus
-
purified enzyme
33.3
-
purified enzyme
40.8
Cellvibrio gilvus
-
purified enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 7
Clostridium stercorarium
-
6.5
-
with D-xylose and 2-deoxyglucose as acceptor
7.5
-
with D-glucosamine as acceptor
7.6
Cellvibrio gilvus
-
-
8
Cellvibrio gilvus
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
Clostridium stercorarium
50% activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38
Cellvibrio gilvus
-
assay at
65
Clostridium stercorarium
-
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 80
-
very low activity at 20 and 80C
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
72000
Cellvibrio gilvus
-
SDS-PAGE
90000
-
gel filtration
92000
-
SDS-PAGE, native PAGE
93760
-
calculated from DNA-sequence
94320
-
calculated from DNA-sequence
150000
180000
Cellvibrio gilvus
-
gel filtration
187000
-
gel filtration
280000
Cellvibrio gilvus
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
tetramer
Cellvibrio gilvus
-
4 * 72000, gel filtration, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
at 25C using the sitting-drop vapour diffusion method. Cellobiose phosphorylase-SO4 and cellobiose phosphorylase-PO4 structures are determined at 2.0 and 2.1 A resolution to R factor (Rfree) of 17.6% (21.3%) and 18.0% (22%) respectively. The pH values for crystallization are pH 8.2 for cellobiose phosphorylase-PO4 and pH 6.5 for cellobiose phosphorylase-SO4.
Cellvibrio gilvus
-
crystallization of recombinant enzyme by the sitting-drop vapour-diffusion method using 10 mg/ml purified enzyme, 1.5 M ammonium sulfate, 0.1 M MES buffer pH 7.0 and 5 mM glucose. A suitable crystal is obtained after 10 d incubation at 298 K. The crystal belongs to space group P2(1), with unit-cell parameters a = 84.77, b = 98.31, c = 104.04 A, beta = 102.73. X-ray diffraction data to 2.1 A resolution collected at KEK-PF BL-5A
Cellvibrio gilvus
-
in complex with phosphate, to 2.4 A resolution. Enzyme is composed of four distinct domains, an N-terminal domain with residues 1-279, a helical linker with residues 280-314, an (alpha/beta)6-barrel domain with residues 321-734, and a C-terminal beta-sandwich domain with residues 315-320 and 735-811. The side chains of His653, Gln699 and Thr718 and the backbone N-atom of Gly719 coordinate the phosphate bound in the interior of the (alpha/beta)6-barrel domain. Two short helical extensions, residues 158-169, from the N-terminal domain of the adjacent subunit extend into the cleft which runs along the face of the (alpha/beta)6-barrel domain, forming an active-site pocket. An additional loop, residues 488-507, lays on top of the active-site pocket, forming a restrictive active-site pocket that is large enough to bind a disaccharide but not large enough to bind an oligosaccharide
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 9
-
most stable at pH 7, rapid loss of activity at different pH
637863
5.5 - 8.8
-
stable within
719128
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
85
O52504
active for 2 h in presence of substrate, without substrate little activity after 15 min
90
-
25% loss of activity after heating for 30 min at pH 6-8
100
-
complete loss of activity after heating for 30 min at pH 6-8
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
storage of purified enzyme at -20C without thiol reagents causes rapid loss of activity
Cellvibrio gilvus
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ni-NTA agarose slurry, Superdex 200 column and Mono Q column
-
recombinant enzyme
-
recombinant enzyme from E. coli
recombinant enzyme with a cellobiose-binding intein tag, development of a simple, low-cost, and scalable protein purification method using a biodegradable regenerated amorphous cellulose, affinity adsorption on regenerated amorphous cellulose followed by intein self-cleavage, method evaluation, overview
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by immobilized metal ion affinity chromatography
use of thiol reagent necessary during purification to maintain activity
Cellvibrio gilvus
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
CP gene, expression of His-tagged wild-type and mutant enzymes in Escherichia coli
expressed in Escherichia coli
expressed in Escherichia coli BL21 (DE3)
Cellvibrio gilvus
-
expressed in Escherichia coli strain BL21 (DE3)
-
expressed in Escherichia coli, part of oligosaccharide catabolic cluster
O52504
expression in Escherichia coli
expression of a functional recombinant enzyme containing an N-terminal metal affinity fusion peptide in Escherichia coli. Catalytic properties of the natural enzyme are retained completely in the recombinant cellobiose phosphorylase
-
expression of intein-tagged enzyme in Escherichia coli strain BL21, subcloning in Escherichia coli strain DH5alpha
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A397V
random, site-saturation, and site-directed mutagenesis, the mutant enzyme shows reduced activity with lactose compared to the wild-type enzyme
A397V/T508A/A512T/D557N/N667T/G681S
random, site-saturation, and site-directed mutagenesis, the mutant enzyme shows 3fold increased activity with lactose and 6fold decreased activity with cellobiose compared to the wild-type enzyme
A512T
random, site-saturation, and site-directed mutagenesis, the mutant enzyme shows similar activity with lactose compared to the wild-type enzyme
D557N
random, site-saturation, and site-directed mutagenesis, the mutant enzyme shows similar activity with lactose compared to the wild-type enzyme
E649C
-
mutant accepts methyl beta-D-glucoside and ethyl beta-D-glucoside as substrates
G681S
random, site-saturation, and site-directed mutagenesis, the mutant enzyme shows reduced activity with lactose compared to the wild-type enzyme
N156D/N163D/T508I/E649G/N667A
-
mutant with very broad acceptor specificity
T508A/N667T
random, site-saturation, and site-directed mutagenesis, the mutant enzyme shows increased activity with lactose compared to the wild-type enzyme
T508I/E649C/N667A
-
triple mutation broadens acceptor specificity
T508I/N667A
A423S
-
mutation introduced to increase thermostability, mutant displays a half-life of 11.3 min at 70C
A781K
-
mutation introduced to increase thermostability, mutant displays a half-life of 15.3 min at 70C
Q130H/K131Y/S411G/A423S/A781K
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mutations introduced to increase thermostability, mutant displays a half-life of 17.7 min at 70C
R48R/Q130H/K131Y/K142R/S411G/A423S/V526A/A781K
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mutations introduced to increase thermostability, mutant displays a half-life of 24.6 min at 70C
S411G
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mutation introduced to increase thermostability, mutant displays a half-life of 13.6 min at 70C
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
Cellvibrio gilvus
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quantitative determination of cellobiose in presence of glucose or glucose-1-phosphate
synthesis