Information on EC 2.4.1.20 - cellobiose phosphorylase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
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
2.4.1.20
-
RECOMMENDED NAME
GeneOntology No.
cellobiose phosphorylase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
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
computational analyses to elucidate the conformational itinerary along the reaction pathway of this enzyme using the substrate binding structure determined from crystal structure, predicted reaction pathway, overview
-
cellobiose + phosphate = alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
reaction proceeds through a random-ordered bi-bi mechanism, by which phosphate and cellobiose bind in random order and D-glucose is released before glucose 1-phosphate
-
cellobiose + phosphate = alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
reaction proceeds through a random-ordered bi-bi mechanism, by which phosphate and cellobiose bind in random order and D-glucose is released before glucose 1-phosphate
Ruminococcus albus NE1
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hexosyl group transfer
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Starch and sucrose metabolism
-
SYSTEMATIC NAME
IUBMB Comments
cellobiose:phosphate alpha-D-glucosyltransferase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Cbp
-
cellobiose phosphorylase
cellobiose:orthophosphate alpha-D-glucosyltransferase
-
-
-
-
CPase
Clostridium thermocellum YM4
Q8VP44
-
-
GH94 cellobiose phosphorylase
-
-
lactose phosphorylase
Q7WTR6
created through directed evolution
CAS REGISTRY NUMBER
COMMENTARY
9030-20-0
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
; DSM 20108
-
-
Manually annotated by BRENDA team
DSM 20108
UniProt
Manually annotated by BRENDA team
strain NCIB 11754
SwissProt
Manually annotated by BRENDA team
Clostridium stercorarium NCIB 11754
strain NCIB 11754
SwissProt
Manually annotated by BRENDA team
member of glycoside hydrolase family 94
UniProt
Manually annotated by BRENDA team
strain ATCC 27405
-
-
Manually annotated by BRENDA team
strain YM4 (AB013109). To convert cellobiose into amylose: the cellobiose phosphorylase catalyzes phosphorolysis of cellobiose into G1-P and glucose, then alpha-glucan phosphorylase catalyses the polymerization of alpha-1,4-glucan, using G1-P as a carbohydrate donor and maltotetraose as the initial carbohydrate acceptor. Thus, glucosyl moieties of cellobiose molecules are successively transferred to the non-reducing ends of glucan to elongate the chain. The optimal conditions are: thirty milligrams per milliliters (87.7 mM) cellobiose, 30 mM sodium phosphate (pH 7.0), 30 microg/ml cellobiose phosphorylase, 30 microg/ml alpha-glucan phosphorylase, and 75 microM maltotetraose are incubated at 45C
-
-
Manually annotated by BRENDA team
Clostridium thermocellum YM4
strain YM4
-
-
Manually annotated by BRENDA team
Clostridium thermocellum YM4
strain YM4
UniProt
Manually annotated by BRENDA team
Clostridium thermocellum YM4
strain YM4 (AB013109). To convert cellobiose into amylose: the cellobiose phosphorylase catalyzes phosphorolysis of cellobiose into G1-P and glucose, then alpha-glucan phosphorylase catalyses the polymerization of alpha-1,4-glucan, using G1-P as a carbohydrate donor and maltotetraose as the initial carbohydrate acceptor. Thus, glucosyl moieties of cellobiose molecules are successively transferred to the non-reducing ends of glucan to elongate the chain. The optimal conditions are: thirty milligrams per milliliters (87.7 mM) cellobiose, 30 mM sodium phosphate (pH 7.0), 30 microg/ml cellobiose phosphorylase, 30 microg/ml alpha-glucan phosphorylase, and 75 microM maltotetraose are incubated at 45C
-
-
Manually annotated by BRENDA team
Fomes annosus
-
-
-
Manually annotated by BRENDA team
Ruminococcus albus NE1
-
-
-
Manually annotated by BRENDA team
strain MSB8
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,5-anhydroglucitol + glucose-1-phosphate
?
show the reaction diagram
-
-
-
-
?
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
Ruminococcus albus, Ruminococcus albus NE1
-
-
-
-
r
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
?
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
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + 2-deoxy-D-glucose
?
show the reaction diagram
-
poor substrate
-
-
?
alpha-D-glucose 1-phosphate + 2-deoxy-D-glucose
?
show the reaction diagram
Clostridium thermocellum YM4
-
-
-
-
?
alpha-D-glucose 1-phosphate + 2-deoxy-D-glucose
? + phosphate
show the reaction diagram
Ruminococcus albus, Ruminococcus albus NE1
-
-
-
-
r
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
-
very poor substrate
-
-
?
alpha-D-glucose 1-phosphate + 5a-carba-beta-D,L-glucopyranose
?
show the reaction diagram
-
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
?
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
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + 6-deoxy-D-glucose
?
show the reaction diagram
-
much better substrate than 2-deoxy-D-glucose
-
-
?
alpha-D-glucose 1-phosphate + 6-deoxy-D-glucose
?
show the reaction diagram
Clostridium thermocellum YM4
-
-
-
-
?
alpha-D-glucose 1-phosphate + 6-deoxy-D-glucose
? + phosphate
show the reaction diagram
Ruminococcus albus, Ruminococcus albus NE1
-
-
-
-
r
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-altrose
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + D-arabinose
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + D-arabinose
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + D-arabinose
?
show the reaction diagram
-
very poor substrate
-
-
?
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
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + D-glucosamine
?
show the reaction diagram
-
-
-
-
-
alpha-D-glucose 1-phosphate + D-glucosamine
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + D-glucosamine
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + D-glucosamine
?
show the reaction diagram
-
poor substrate
-
-
?
alpha-D-glucose 1-phosphate + D-glucosamine
? + 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-glucose
cellobiose + phosphate
show the reaction diagram
-
-
-
-
r
alpha-D-glucose 1-phosphate + D-glucose
cellobiose + phosphate
show the reaction diagram
-
-
-
-
r
alpha-D-glucose 1-phosphate + D-glucose
cellobiose + phosphate
show the reaction diagram
Ruminococcus albus, Ruminococcus albus NE1
-
in synthesis reaction, glucose is best substrate
-
-
r
alpha-D-glucose 1-phosphate + D-glucose
? + phosphate
show the reaction diagram
-
-
-
-
r
alpha-D-glucose 1-phosphate + D-mannose
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + D-mannose
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + D-mannose
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + D-mannose
?
show the reaction diagram
-
-
-
-
-
alpha-D-glucose 1-phosphate + D-mannose
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + D-mannose
?
show the reaction diagram
-
poor substrate
-
-
?
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
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + D-xylose
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + D-xylose
?
show the reaction diagram
-
poor substrate
-
-
?
alpha-D-glucose 1-phosphate + D-xylose
?
show the reaction diagram
-
no activity with L-isomer
-
-
?
alpha-D-glucose 1-phosphate + D-xylose
?
show the reaction diagram
-
no activity with L-isomer
-
-
?
alpha-D-glucose 1-phosphate + D-xylose
?
show the reaction diagram
-
higher specific activity than with D-glucose as acceptor
-
-
?
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-fucose
?
show the reaction diagram
-
-
-
-
?
alpha-D-glucose 1-phosphate + L-galactose
?
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
-
-
-
-
r
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
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
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
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
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
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
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
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
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
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Fomes annosus
-
-
-
-
-
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Fomes annosus
-
-
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-, Q59316
-
-
-
-
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-, Q59316
-
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
O52504, -
-
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-, Q7WTR6
-
-
-
r
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Q8VP44
-
-
-
r
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-
ordered bi-bi mechanism
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-
beta-cellobiose preferred over alpha-anomer
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-
beta-cellobiose preferred over alpha-anomer
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-
random ordered bi-bi mechanism
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-
beta-D-cellobiose is phosphorylyzed faster than alpha-anomer, ordered bi-bi mechanism
beta-D-glucose was shown to be the 4 times better acceptor compared to alpha-D-glucose in the reverse reaction
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-
sequential bi-bi mechanism
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-
sequential bi-bi mechanism
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-
ternary-complex kinetic mechanism
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
O52504, -
nonconventional pathway for glucan utilization
-
-
-
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-
important for energy efficient catabolism of cellobiose in cytoplasm
-
-
-
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-
CBP catalyzes the phosphorolysis of cellobiose into alpha-D-glucose 1-phosphate and D-glucose with inversion of anomeric configuration, computational docking, overview
-
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Q8VP44
the enzyme shows beta-anomer selective glucosylation activity towards disaccharides
-
-
r
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Ruminococcus albus NE1
-
-
-
-
r
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Clostridium thermocellum YM4
-
-
-
-
-
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Clostridium thermocellum YM4
-
ordered bi-bi mechanism, beta-cellobiose preferred over alpha-anomer
-
?
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Clostridium thermocellum YM4
Q8VP44
-, the enzyme shows beta-anomer selective glucosylation activity towards disaccharides
-
-
r
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Clostridium stercorarium NCIB 11754
Q59316
-
-
-
-
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Clostridium stercorarium NCIB 11754
Q59316
-
-
?
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
-
-
-
-
r
D-glucal + D-glucose
2-deoxy-beta-D-arabino-hexopyranosyl-(1->4)-D-glucose + ?
show the reaction diagram
-
-
-
-
r
D-glucal + D-mannose
2-deoxy-beta-D-arabino-hexopyranosyl-(1->4)-D-mannose + ?
show the reaction diagram
-
-
-
-
r
D-glucal + D-xylose
2-deoxy-beta-D-arabino-hexopyranosyl-(1->4)-D-xylose + ?
show the reaction diagram
-
-
-
-
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
-
-
-
-
?
isomaltose + alpha-D-glucose 1-phosphate
?
show the reaction diagram
-
-
-
-
?
lactose + phosphate
alpha-D-galactose 1-phosphate + D-glucose
show the reaction diagram
-, Q7WTR6
-
-
-
r
melibiose + alpha-D-glucose 1-phosphate
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
no activity with D-xylose and D-mannose
-
-
-
additional information
?
-
-, Q59316
only disaccharide accepted as substrate
-
-
-
additional information
?
-
O52504, -
only disaccharide accepted as substrate
-
-
-
additional information
?
-
-
no activity detected with glucose-acceptor different from D-glucose
-
-
-
additional information
?
-
Clostridium thermocellum, Clostridium thermocellum YM4
Q8VP44
beta-glucosylation of alkyls by the enzyme using either cellobiose or alpha-D-glucose 1-phosphate as donors, active with methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, tert-butanol, n-pentanol, n-hexanol, n-heptanol, 1,2-butanediol, 1,3-butanediol, and 1,4-butanediol
-
-
-
additional information
?
-
Clostridium stercorarium NCIB 11754
Q59316
only disaccharide accepted as substrate
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
alpha-D-glucose 1-phosphate + D-glucose
cellobiose + phosphate
show the reaction diagram
-
-
-
-
r
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
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
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
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Cellulomonas sp., Fomes annosus
-
-
-
-
-
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-, Q59316
-
-
-
-
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-, Q7WTR6
-
-
-
r
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
O52504, -
nonconventional pathway for glucan utilization
-
-
-
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
-
important for energy efficient catabolism of cellobiose in cytoplasm
-
-
-
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Q8VP44
the enzyme shows beta-anomer selective glucosylation activity towards disaccharides
-
-
r
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Clostridium thermocellum YM4
-
-
-
-
-
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Clostridium thermocellum YM4
Q8VP44
the enzyme shows beta-anomer selective glucosylation activity towards disaccharides
-
-
r
cellobiose + phosphate
alpha-D-glucose 1-phosphate + D-glucose
show the reaction diagram
Clostridium stercorarium NCIB 11754
Q59316
-
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mg2+
-
required
Mg2+
-
not required
additional information
-
no influence of metal ions on reaction
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
substrate inhibition
alpha-oxogluconate
-
-
cellobiose
-
substrate inhibition, competitive with glucose-1-phosphate
D-cellobiose
-
-
D-glucosamine
-
substrate inhibition
D-glucose
-
product inhibition
D-glucose
-
substrate inhibition, competitive with glucose-1-phosphate
D-glucose
-
strong inhibition at high concentrations
D-glucose
-
mixed type inhibition against cellobiose, pure beta-D-glucose inhibits stronger than mixture of both anomers
D-glucose
-
substrate inhibition, competitive with glucose-1-phosphate
D-glucose
-
substrate inhibition, competitive with glucose-1-phosphate
D-glucose
-
substrate inhibition
D-glucose
-
inhibition of the synthetic reaction, binds competitively at the glucose 1-phosphate site and uncompetitvely at the ternary enzyme-glucose 1-phosphate-glucose complex
D-glucose 1-phosphate
-
substrate inhibition, competitive with cellobiose
D-glucose 1-phosphate
-
-
glucono-delta-lactone
-
very weak
N-ethylmaleimide
-
56% inhibition at 0.5 mM
Nojirimycin
-
competitive with respect to cellobiose
p-chloromercuribenzoate
-
complete inhibition at 0.02 mM
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
9.5
-
1,5-Anhydro-D-glucitol
-
pH 6.0, 37C
8
9
1,5-anhydroglucitol
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
17.2
-
2-deoxy-2-fluoro-D-glucose
-
-
24.3
-
2-deoxy-D-glucose
-
-
47
-
2-deoxy-D-glucose
-
-
56.9
-
2-deoxy-D-glucose
-
-
60.2
-
2-deoxy-D-glucose
-
pH 6.0, 37C
73
-
2-deoxy-D-glucose
-
-
168
-
2-deoxy-D-glucose
-
-
250
-
2-deoxy-D-glucose
-
-
7.4
-
6-deoxy-6-fluoro-D-glucose
-
-
4.1
-
6-deoxy-D-glucose
-
-
6.1
-
6-deoxy-D-glucose
-
-
9.2
-
6-deoxy-D-glucose
-
-
9.8
-
6-deoxy-D-glucose
-
pH 6.0, 37C
10.2
-
6-deoxy-D-glucose
-
-
24
-
6-deoxy-D-glucose
-
-
2.15
-
alpha-D-glucose 1-phosphate
-
-
2.15
-
alpha-D-glucose 1-phosphate
-
pH 6.6, 30C, native enzyme
3.28
-
alpha-D-glucose 1-phosphate
-
pH 6.6, 30C, recombinant His6-tagged enzyme
8
-
alpha-D-glucose 1-phosphate
-
-
33.7
-
butyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
0.29
-
cellobiose
-
-
1.25
-
cellobiose
-
-
1.42
-
cellobiose
O52504, -
-
2.7
-
cellobiose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant T508I/N667A
2.9
-
cellobiose
-
pH 6.6, 30C, recombinant His6-tagged enzyme
2.9
-
cellobiose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant T508A/N667T; pH 6.6, 37C, recombinant wild-type enzyme
3.3
-
cellobiose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant A397V/T508A/A512T/D557N/N667T/G681S
4.3
-
cellobiose
-
-
4.3
-
cellobiose
-
pH 6.6, 30C, native enzyme
6.2
-
cellobiose
-, Q59316
-
6.9
-
cellobiose
-
mutant S411G, pH 7.2, 40C
10.3
-
cellobiose
-
mutant A423S, pH 7.2, 40C
10.8
-
cellobiose
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
11
-
cellobiose
-
mutant A781 K, pH 7.2, 40C
14.7
-
cellobiose
-
mutant R48R/Q130H/K131Y/K142R/S411G/A423S/V526A/A781K, pH 7.2, 40C
15.9
-
cellobiose
-
mutant Q130H/K131Y/S411G/A423S/A781K, pH 7.2, 40C
16.5
-
cellobiose
-
wild-type, pH 7.2, 40C
240
-
D-arabinose
-
-
2.6
-
D-cellobiose
-
-
10.8
-
D-gentobiose
-
pH 6.0, 37C
-
5.7
-
D-glucosamine
-
-
5.9
-
D-glucosamine
-
-
9.5
-
D-glucosamine
-
-
10
-
D-glucosamine
-
-
13
-
D-glucosamine
-
-
13.3
-
D-glucosamine
-
pH 6.0, 37C
0.69
-
D-glucose
-
-
1.46
-
D-glucose
-
pH 6.0, 37C
2.1
-
D-glucose
-
-
2.26
-
D-glucose
-
-
2.26
-
D-glucose
-
pH 6.6, 30C, native enzyme
3.79
-
D-glucose
-
pH 6.6, 30C, recombinant His6-tagged enzyme
17.2
-
D-glucose
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
28.9
-
D-glucose
-
mutant T508I/E649C/N667A, pH 6.6, 37C
10.8
-
D-glucoseamine
-
-
-
23.5
-
D-mannose
-
pH 6.0, 37C
27.3
-
D-mannose
-
-
67
-
D-mannose
-
-
115
-
D-mannose
-
-
14
-
D-xylose
-
-
15.9
-
D-xylose
-
-
25.5
-
D-xylose
-
pH 6.0, 37C
84
-
D-xylose
-
-
138
-
ethyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
-
155
-
ethyl beta-D-glucoside
-
mutant T508I/E649C/N667A, pH 6.6, 37C
-
83
-
Gentiobiose
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
71
-
isomaltose
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
228
-
lactose
-, Q7WTR6
pH 6.6, 37C, recombinant wild-type enzyme
230
-
lactose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant T508I/N667A
254
-
lactose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant T508A/N667T
330
-
lactose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant A397V/T508A/A512T/D557N/N667T/G681S
93
-
melibiose
-
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
-
methyl beta-D-glucoside
-
mutant T508I/E649C/N667A, pH 6.6, 37C
31.2
-
phenyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
0.15
-
phosphate
-
-
0.61
-
phosphate
-
-
0.77
-
phosphate
-
-
0.82
-
phosphate
-
pH 6.6, 30C, recombinant His6-tagged enzyme
1.2
-
phosphate
-
-
1.2
-
phosphate
-
pH 6.6, 30C, native enzyme
79.8
-
methyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
additional information
-
additional information
-
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]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2.8
-
1,5-Anhydro-D-glucitol
-
pH 6.0, 37C
3.5
-
1,5-anhydroglucitol
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
8
-
2-deoxy-2-fluoro-D-glucose
-
-
11.6
-
2-deoxy-D-glucose
-
-
16
-
2-deoxy-D-glucose
-
-
18.3
-
2-deoxy-D-glucose
-
-
21.5
-
2-deoxy-D-glucose
-
pH 6.0, 37C
44.6
-
6-deoxy-6-fluoro-D-glucose
-
-
17
-
6-deoxy-D-glucose
-
-
55.3
-
6-deoxy-D-glucose
-
-
59.5
-
6-deoxy-D-glucose
-
-
124
-
6-deoxy-D-glucose
-
pH 6.0, 37C
43.2
-
alpha-D-glucose 1-phosphate
-
pH 6.6, 30C, native enzyme; pH 6.6, 30C, recombinant His6-tagged enzyme
9
-
butyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
1.1
-
cellobiose
-, Q59316
-
1.3
-
cellobiose
-
mutant S411G, pH 7.2, 40C
1.5
-
cellobiose
-
mutant R48R/Q130H/K131Y/K142R/S411G/A423S/V526A/A781K, pH 7.2, 40C
2.01
-
cellobiose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant T508I/N667A
2.1
-
cellobiose
-
mutant Q130H/K131Y/S411G/A423S/A781K, pH 7.2, 40C
2.6
-
cellobiose
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
2.61
-
cellobiose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant T508A/N667T
2.9
-
cellobiose
-
mutant A423S, pH 7.2, 40C; mutant A781 K, pH 7.2, 40C
3.28
-
cellobiose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant A397V/T508A/A512T/D557N/N667T/G681S
3.4
-
cellobiose
-
wild-type, pH 7.2, 40C
5.4
-
cellobiose
-
-
13.2
-
cellobiose
-, Q7WTR6
pH 6.6, 37C, recombinant wild-type enzyme
29.1
-
cellobiose
-
-
29.1
-
cellobiose
-
pH 6.6, 30C, native enzyme; pH 6.6, 30C, recombinant His6-tagged enzyme
4
-
D-gentobiose
-
pH 6.0, 37C
-
5.2
-
D-glucosamine
-
-
7.7
-
D-glucosamine
-
-
9.56
-
D-glucosamine
-
pH 6.0, 37C
5.3
-
D-glucose
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
6.3
-
D-glucose
-
mutant T508I/E649C/N667A, pH 6.6, 37C
8
-
D-glucose
-
-
27.5
-
D-glucose
-
pH 6.6, 30C, native enzyme; pH 6.6, 30C, recombinant His6-tagged enzyme
43.2
-
D-glucose
-
-
92.3
-
D-glucose
-
pH 6.0, 37C
5.6
-
D-glucoseamine
-
-
-
2.83
-
D-mannose
-
pH 6.0, 37C
4.4
-
D-mannose
-
-
6.6
-
D-mannose
-
-
8.7
-
D-xylose
-
-
16.5
-
D-xylose
-
pH 6.0, 37C
40
-
D-xylose
-
-
5.1
-
ethyl beta-D-glucoside
-
mutant T508I/E649C/N667A, pH 6.6, 37C
-
8.7
-
ethyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
-
7.2
-
Gentiobiose
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
1.9
-
isomaltose
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
0.109
-
lactose
-, Q7WTR6
pH 6.6, 37C, recombinant wild-type enzyme
0.35
-
lactose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant T508I/N667A
0.53
-
lactose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant T508A/N667T
1.11
-
lactose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant A397V/T508A/A512T/D557N/N667T/G681S
2.3
-
melibiose
-
enzyme assay at 37C in 50 mM Tris/HCl (pH 7.0)
6
-
methyl alpha-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
6.8
-
methyl beta-D-glucoside
-
mutant T508I/E649C/N667A, pH 6.6, 37C
7.9
-
methyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
4
-
phenyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
15.4
-
phosphate
-
pH 6.6, 30C, native enzyme; pH 6.6, 30C, recombinant His6-tagged enzyme
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.29
-
1,5-Anhydro-D-glucitol
-
pH 6.0, 37C
769
0.36
-
2-deoxy-D-glucose
-
pH 6.0, 37C
2416
12.7
-
6-deoxy-D-glucose
-
pH 6.0, 37C
5639
0.266
-
butyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
8140
0.103
-
cellobiose
-
mutant R48R/Q130H/K131Y/K142R/S411G/A423S/V526A/A781K, pH 7.2, 40C
8399
0.134
-
cellobiose
-
mutant Q130H/K131Y/S411G/A423S/A781K, pH 7.2, 40C
8399
0.189
-
cellobiose
-
mutant S411G, pH 7.2, 40C
8399
0.2
-
cellobiose
-
wild-type, pH 7.2, 40C
8399
0.24
-
cellobiose
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
8399
0.263
-
cellobiose
-
mutant A781 K, pH 7.2, 40C
8399
0.287
-
cellobiose
-
mutant A423S, pH 7.2, 40C
8399
0.9
-
cellobiose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant T508A/N667T
8399
0.99
-
cellobiose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant A397V/T508A/A512T/D557N/N667T/G681S
8399
1.21
-
cellobiose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant T508I/N667A
8399
4.56
-
cellobiose
-, Q7WTR6
pH 6.6, 37C, recombinant wild-type enzyme
8399
0.37
-
D-gentobiose
-
pH 6.0, 37C
0
0.72
-
D-glucosamine
-
pH 6.0, 37C
9197
0.031
-
D-glucose
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
9202
0.217
-
D-glucose
-
mutant T508I/E649C/N667A, pH 6.6, 37C
9202
63.2
-
D-glucose
-
pH 6.0, 37C
9202
0.12
-
D-mannose
-
pH 6.0, 37C
9286
0.65
-
D-xylose
-
pH 6.0, 37C
9403
0.033
-
ethyl beta-D-glucoside
-
mutant T508I/E649C/N667A, pH 6.6, 37C
0
0.063
-
ethyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
0
0.00048
-
lactose
-, Q7WTR6
pH 6.6, 37C, recombinant wild-type enzyme
12479
0.0015
-
lactose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant T508I/N667A
12479
0.0021
-
lactose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant T508A/N667T
12479
0.0034
-
lactose
-, Q7WTR6
pH 6.6, 37C, recombinant mutant A397V/T508A/A512T/D557N/N667T/G681S
12479
0.006
-
methyl alpha-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
76411
0.099
-
methyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
13044
0.12
-
methyl beta-D-glucoside
-
mutant T508I/E649C/N667A, pH 6.6, 37C
13044
0.129
-
phenyl beta-D-glucoside
-
mutant N156D/N163D/T508I/E649G/N667A, pH 6.6, 37C
15403
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
-
1.2
-
D-glucose
-
product inhibition
2.2
-
D-glucose
-
-
202
-
D-glucose
-
pH 6.6, 30C, substrate inhibition, native enzyme
255
-
D-glucose
-
pH 6.6, 30C, substrate inhibition, recombinant His6-tagged enzyme
3.2
-
D-glucose 1-phosphate
-
-
0.045
-
Nojirimycin
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.033
-
-, Q7WTR6
purifed recombinant wild-type enzyme, substrate lactose
0.1
-
-, Q7WTR6
purifed recombinant mutant A397V/T508A/A512T/D557N/N667T/G681S, substrate lactose
0.171
-
-, Q7WTR6
purifed recombinant mutant T508I/N667A, substrate lactose
0.249
-
-, Q7WTR6
purifed recombinant mutant T508A/N667T, substrate lactose
1.35
-
-, Q7WTR6
purifed recombinant mutant A397V/T508A/A512T/D557N/N667T/G681S, substrate cellobiose
1.71
-
-, Q7WTR6
purifed recombinant mutant T508I/N667A, substrate cellobiose
2.61
-
-, Q7WTR6
purifed recombinant mutant T508A/N667T, substrate cellobiose
2.66
-
-
purified enzyme
4.7
-
-, Q59316
purified enzyme
7.78
-
-, Q7WTR6
purifed recombinant wild-type enzyme, substrate cellobiose
11.8
-
O52504, -
-
14.4
-
-
pH 6.6, 30C
20
-
-
purified enzyme
27.4
-
-
purified enzyme
33.3
-
-
purified enzyme
40.8
-
-
purified enzyme
additional information
-
-
-
additional information
-
-
the cellobiose phosphorylase activites in the concentrated cell extract is 84 IU/ml.
additional information
-
-
the activity on D-glucal is approximately 500times less than that on alpha-D-glucose 1-phosphate at 10 mM of each donor with 10 mM D-glucose by comparing the initial rate of the production
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
-
-
with D-xylose and 2-deoxyglucose as acceptor
6.6
-
-, Q7WTR6
assay at
7
-
-
forward and reverse reaction
7
-
Q8VP44
assay at
7.5
-
-
with D-glucosamine as acceptor
8
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
8
-, Q59316
50% activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-, Q7WTR6
assay at
38
-
-
assay at
40
-
Q8VP44
assay at
60
-
-
wild-type
80
-
-
mutant R48R/Q130H/K131Y/K142R/S411G/A423S/V526A/A781K
85
-
O52504, -
-
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
80
-
very low activity at 20 and 80C
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
72000
-
-
SDS-PAGE
85000
-
-
SDS-PAGE
85000
-
-
SDS-PAGE
85000
-
-
SDS-PAGE
90000
-
-
gel filtration
92000
-
-
SDS-PAGE, native PAGE
93000
-
-, Q59316
SDS-PAGE, gel filtration
93000
-
O52504, -
SDS-PAGE
93760
-
-
calculated from DNA-sequence
94320
-
-
calculated from DNA-sequence
150000
-
-
gel filtration
150000
-
-
gel filtration
180000
-
-
gel filtration
187000
-
-
gel filtration
280000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dimer
-
2 * 85000, gel filtration, SDS-PAGE
dimer
-
2 * 85000, gel filtration, SDS-PAGE
dimer
-
2 * 85000, gel filtration, SDS-PAGE
dimer
-
2 * 94000, SDS-PAGE
dimer
Ruminococcus albus NE1
-
2 * 94000, SDS-PAGE
-
monomer
-, Q59316
1 * 93000, SDS-PAGE, gel filtration
monomer
-
1 * 92000, SDS-PAGE, native PAGE
monomer
-
1 * 92000, SDS-PAGE
monomer
Clostridium stercorarium NCIB 11754
-
1 * 93000, SDS-PAGE, gel filtration
-
monomer
Clostridium thermocellum YM4
-
1 * 92000, SDS-PAGE, native PAGE
-
tetramer
-
4 * 72000, gel filtration, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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.
-
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
-
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
-, Q8VP44
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
9
-
most stable at pH 7, rapid loss of activity at different pH
5.5
8.8
-
stable within
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
40
-
-
inactivated above
40
-
-
stable up to
50
-
-
unstable above
60
-
-, Q59316
stable for at least 42 h
60
-
-
complete inactivation within 10 min
70
-
-
less than 10% loss of activity after heating for 30 min at pH 6-8
70
-
-
half-life of wild-type 8.3 min, mutant Q130H/K131Y/S411G/A423S/A781K 17.7 min, mutant R48R/Q130H/K131Y/K142R/S411G/A423S/V526A/A781K 24.6 min
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 ACCESSION NO.
LITERATURE
storage of purified enzyme at -20C without thiol reagents causes rapid loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant enzyme
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by immobilized metal ion affinity chromatography
-, Q7WTR6
use of thiol reagent necessary during purification to maintain activity
-
Ni-NTA agarose slurry, Superdex 200 column and Mono Q column
-
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 enzyme from E. coli
O52504, -
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
CP gene, expression of His-tagged wild-type and mutant enzymes in Escherichia coli
-, Q7WTR6
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
-
expressed in Escherichia coli
-
expressed in Escherichia coli BL21 (DE3)
-
expression in Escherichia coli
O66264
expressed in Escherichia coli
-
expressed in Escherichia coli strain BL21 (DE3)
-
expression in Escherichia coli
-
expression of intein-tagged enzyme in Escherichia coli strain BL21, subcloning in Escherichia coli strain DH5alpha
-
expression in Escherichia coli
-
expressed in Escherichia coli
-
expressed in Escherichia coli, part of oligosaccharide catabolic cluster
O52504, -
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
A397V
-, Q7WTR6
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
-, Q7WTR6
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
-, Q7WTR6
random, site-saturation, and site-directed mutagenesis, the mutant enzyme shows similar activity with lactose compared to the wild-type enzyme
D557N
-, Q7WTR6
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
N156D/N163D/T508I/E649G/N667A
-
mutant with very broad acceptor specificity
T508A/N667T
-, Q7WTR6
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
-, Q7WTR6
random, site-saturation, and site-directed mutagenesis, the mutant enzyme shows 7.5fold increased activity with lactose compared to the wild-type enzyme
T508I/N667A
-
mutant accepts methyl beta-D-glucoside and ethyl beta-D-glucoside as substrates
A423S
-
mutation introduced to increase thermostability, mutant displays a half-life of 11.3 min at 70C
Q130H/K131Y/S411G/A423S/A781K
-
mutations introduced to increase thermostability, mutant displays a half-life of 17.7 min at 70C
R48R/Q130H/K131Y/K142R/S411G/A423S/V526A/A781K
-
mutations introduced to increase thermostability, mutant displays a half-life of 24.6 min at 70C
S411G
-
mutation introduced to increase thermostability, mutant displays a half-life of 13.6 min at 70C
G681S
-, Q7WTR6
random, site-saturation, and site-directed mutagenesis, the mutant enzyme shows reduced activity with lactose compared to the wild-type enzyme
additional information
-, Q7WTR6
random, site-saturation, and site-directed mutagenesis for directed evolution in order to create enzyme variants with significantly increased lactose phosphorylase activity, useful for the production of alpha-D-galactose 1-phosphate, overview. Location of mutations in the 3D structure, overview
A781K
-
mutation introduced to increase thermostability, mutant displays a half-life of 15.3 min at 70C
additional information
-
method development using the enzyme for production of D-glucose 1-phosphate, which is an expensive substrate for synthesis of amylose by alpha-glucan phosphorylase, overview
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
synthesis
-
ability of the phosphorylase to utilize alpha-D -glucose 1-fluoride as alternate glucosyl donor in place of alpha-D-glucose 1-phosphate for the synthesis of alpha-1,4-glucosides under thermodynamic control in close to 100% yield
analysis
-
quantitative determination of cellobiose in presence of glucose or glucose-1-phosphate
synthesis
-
enzymatic cellobiose synthesis from starch using two phosphorylases: glucan phosphorylase, from Klebsiella pneumoniae, converts glucose residues in the starch into glucose-1-phosphate, removal of phosphate, and glucose-1-phosphate is incubated with alpha-D-glucose and is converted into cellobiose by recombinant cellobiose phosphorylase, removal of phosphate, 60% cellobiose yield from glucose 1-phosphate and, at least, 23.7% cellobiose yield from starch, method optimization, overview
synthesis
O66264
expression of enzyme in Escherichia coli and study on the role of molecular chaperones and growth temperature on the solubilization of enzyme overexpressed in Escherichia coli. The growth of host at low temperature enhances enzyme in soluble fraction. Similarly, induction of target gene at low level of IPTG also yields more enzyme in the soluble fraction. Coexpression of the target gene with molecular chaperones GroESL and KODH does not enhance the solubilization under in vivo conditions
synthesis
-
the cellobiose phosphorylases in the cell extract is used to synthesize radiolabeled cellodextrins with a degree of polymerization (DP=2-6) from non-radioactive glucose-1-phosphate and radioactive glucose. For cellobiose synthesis, the reaction is carried out at 60C for 30 min with crude enzyme. After an impluse feed of radiolabeled cellobiose to a Clostridium thermocellum culture, the intracellular sugar levels are measured: the largest amount of radioactivity is cellobiose with lesser amounts of glucose, cellotriose, and cellotetraose, and an average degree of polymerization of intracellular cellodextrins is ca. 2
synthesis
-
the enzyme is useful for production of D-glucose 1-phosphate, an expensive substrate for other enzymatic syntheses
synthesis
Q8VP44
the enzyme is useful for synthesis of alkyl beta-glucosides, overview
synthesis
Clostridium thermocellum YM4
-
the enzyme is useful for synthesis of alkyl beta-glucosides, overview
-