Any feedback?
Information on EC 2.4.1.230 - kojibiose phosphorylase
for references in articles please use BRENDA:EC2.4.1.230Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
2.4.1.230 kojibiose phosphorylaseIUBMB Comments
The enzyme from Thermoanaerobacter brockii can act with alpha-1,2-oligoglucans, such as selaginose, as substrate, but more slowly. The enzyme is inactive when dissaccharides with linkages other than alpha-1,2 linkages, such as sophorose, trehalose, neotrehalose, nigerose, laminaribiose, maltose, cellobiose, isomaltose, gentiobiose, sucrose and lactose, are used as substrates.
Specify your search results
Word Map
- 2.4.1.230
- brockii
-
thermoanaerobacter
- maltose
- phosphorylases
-
phosphorolysis
- proteinase
-
saccharide
- trehalose
- oligosaccharides
-
keratinolytic
- 1-phosphate
- corneum
- glycoside
- pepstatin
- albicans
-
stratum
- trisaccharide
- nigerose
- synthesis
- saccharolyticus
- chymostatin
-
2d-nmr
- l-sorbose
- dermatophytosis
- beta-d-glucose
- canis
-
scalp
-
caldicellulosiruptor
-
hsqc-tocsy
- microsporum
- brevis
The expected taxonomic range for this enzyme is: Bacteria, Archaea
Synonyms
kpase, kojibiose phosphorylase, py04_1502, 
more
topPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
kojibiose phosphorylase
KPase
-
-
-
-
PsKP
Py04_1502
YcjT
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2-alpha-D-glucosyl-D-glucose + phosphate = D-glucose + beta-D-glucose 1-phosphate
the enzyme from Thermoanaerobacter brockii can act with alpha-1,2-oligoglucans, such as selaginose, as substrate, but more slowly. The enzyme is inactive when dissaccharides with linkages other than alpha-1,2 linkages, such as sophorose, trehalose, neotrehalose, nigerose, laminaribiose, maltose, cellobiose, isomaltose, gentiobiose, sucrose and lactose, are used as substrates
-
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hexosyl group transfer
hexosyl group transfer
-
-
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PATHWAY SOURCE
PATHWAYS
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SYSTEMATIC NAME
IUBMB Comments
2-alpha-D-glucosyl-D-glucose:phosphate beta-D-glucosyltransferase
The enzyme from Thermoanaerobacter brockii can act with alpha-1,2-oligoglucans, such as selaginose, as substrate, but more slowly. The enzyme is inactive when dissaccharides with linkages other than alpha-1,2 linkages, such as sophorose, trehalose, neotrehalose, nigerose, laminaribiose, maltose, cellobiose, isomaltose, gentiobiose, sucrose and lactose, are used as substrates.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CAS REGISTRY NUMBER
COMMENTARY
206566-36-1
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
3-O-alpha-D-glucopyranosyl-D-glucopyranose + phosphate
D-glucose + beta-D-glucose 1-phosphate
low activity, reaction of nigerose phosphorylase, EC 2.4.1.279
-
-
?
alpha,alpha-trehalose + phosphate
D-glucose + beta-D-glucose 1-phosphate
very low activity, reaction of trehalose phosphorylase, EC 2.4.1.64
-
-
?
beta-D-glucose 1-phosphate + D-glucose
2-alpha-D-glucosyl-D-glucose + phosphate
-
-
-
-
?
beta-D-glucose 1-phosphate + glycerol
O-alpha-D-glucopyranosyl-(1,1)-glycerol + O-alpha-glucopyranosyl-(1,2)-O-alpha-D-glucopyranosyl-(1,1)-glycerol + phosphate
-
0.27% of the activity with D-glucose
-
-
?
beta-D-glucose 1-phosphate + myo-inositol
O-alpha-D-glucopyranosyl-(1,1)-myo-inositol + O-alpha-D-glucopyranosyl-(1,5)-myo-inositol + O-alpha-D-glucopyranosyl-(1,2)-O-alpha-D-glucopyranosyl-(1,1)-myo-inositol + O-alpha-D-glucopyranosyl-(1,2)-O-alpha-D-glucopyranosyl-(1,1)-myo-inositol
-
6.2% of the activity with D-glucose
-
-
?
beta-D-glucose 1-phosphate + raffinose
2-alpha-D-glucopyranosyl-raffinose + 2G(2-alpha-D-glucopyranosyl)2-raffinose + 2G(2-alpha-D-glucopyranosyl)3-raffinose
-
-
-
-
?
beta-D-glucose 1-phosphate + stachyose
2-alpha-D-glucopyranosyl-stachyose + 2G(2-alpha-D-glucopyranosyl)2-stachyose + 2G(2-alpha-D-glucopyranosyl)3-stachyose
-
-
-
-
?
cyclo-[(-6)-alpha-D-Glcp-(1-3)-alpha-D-Glcp-(1-6)-alpha-D-Glcp-(1-3)-alpha-D-Glcp-(1-)] + beta-D-glucose 1-phosphate
cyclo-[(-6)-alpha-D-Glcp-(1-3)-alpha-D-Glcp-(1-6)-[alpha-D-Glcp-(1-2)]-alpha-D-Glcp-(1-3)-alpha-D-Glcp-(1-)] + phosphate
-
i.e. CTS
major product, i.e. 2-O-alpha-D-glucopyranosyl-CTS
-
?
D-glucose + O-beta-D-fructofuranosyl-(2-1)-O-beta-D-fructofuranosyl-(2-1)-O-beta-D-fructofuranosyl-(2-1)-glucopyranoside
[O-alpha-D-glucopyranosyl-1(1-2)]n-O-[beta-D-fructofuranosyl-(2-1)]3-alpha-D-glucopyranoside + phosphate
-
-
n = 1 or 2
?
maltose + phosphate
D-glucose + beta-D-glucose 1-phosphate
very low, activity, reaction of maltose phosphorylase, EC 2.4.1.8
-
-
?
O-alpha-D-xylopyranosyl-(1-2)-beta-D-fructofuranoside + 2 beta-D-glucose 1-phosphate
O-alpha-D-glucopyranosyl-(1-[2-O-alpha-D-glucopyranosyl-1]1-2)-O-alpha-D-xylopyranosyl-(1-2)-beta-D-fructofuranoside + 2 phosphate
-
-
-
r
O-alpha-D-xylopyranosyl-(1-2)-beta-D-fructofuranoside + 3 beta-D-glucose 1-phosphate
O-alpha-D-glucopyranosyl-(1-[2-O-alpha-D-glucopyranosyl-1]2-2)-O-alpha-D-xylopyranosyl-(1-2)-beta-D-fructofuranoside + 3 phosphate
-
-
-
r
O-alpha-D-xylopyranosyl-(1-2)-beta-D-fructofuranoside + beta-D-glucose 1-phosphate
O-alpha-D-glucopyranosyl-(1-[2-O-alpha-D-glucopyranosyl-1]0-2)-O-alpha-D-xylopyranosyl-(1-2)-beta-D-fructofuranoside + phosphate
-
-
-
r
palatinose + 2 beta-D-glucose 1-phosphate
2G(2-alpha-D-glucopyranosyl)2-palatinose + 2 phosphate
-
-
-
r
palatinose + 2 beta-D-glucose 1-phosphate
O-alpha-D-glucopyranosyl-(1-2)-O-alpha-D-glucopyranosyl-(1-2)-O-alpha-D-glucopyranosyl-(1-6)-D-fructofuranose + 2 phosphate
-
-
-
r
palatinose + beta-D-glucose 1-phosphate
2G-alpha-D-glucopyranosyl-palatinose + phosphate
-
-
-
r
palatinose + beta-D-glucose 1-phosphate
O-alpha-D-glucopyranosyl-(1-2)-O-alpha-D-glucopyranosyl-(1-6)-D-fructofuranose + phosphate
-
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
?
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
?
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
activity of a LaMP loop 3 maltose phosphorylase mutant, no activity with wild-type maltose phosphorylase, which is strictly specific for maltose
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
activity of a LaMP loop 3 maltose phosphorylase mutant, no activity with wild-type maltose phosphorylase, which is strictly specific for maltose
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
2-alpha-D-glucosyl-D-glucose i.e. kojibiose. The enzyme is involved in conversion of kojibiose into beta-D-glucose 6-phosphate, a substrate of the glycolytic pathway. Disaccharide utilization pathway via phosphorolysis in hyperthermophilic archaea
-
-
?
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
2-alpha-D-glucosyl-D-glucose i.e. kojibiose
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
2-alpha-D-glucosyl-D-glucose i.e. kojibiose. The enzyme is involved in conversion of kojibiose into beta-D-glucose 6-phosphate, a substrate of the glycolytic pathway. Disaccharide utilization pathway via phosphorolysis in hyperthermophilic archaea
-
-
?
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
2-alpha-D-glucosyl-D-glucose i.e. kojibiose
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
-
?
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
-
?
3-O-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
1-alpha-D-glucosyl-D-glucose i.e. nigerose, at 0.23% of the activity with kojibiose
-
-
?
3-O-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
1-alpha-D-glucosyl-D-glucose i.e. nigerose, at 0.23% of the activity with kojibiose
-
-
?
beta-D-glucose + beta-D-glucose 1-phosphate
2-alpha-D-glucosyl-D-glucose + phosphate
-
-
-
-
r
beta-D-glucose + beta-D-glucose 1-phosphate
2-alpha-D-glucosyl-D-glucose + phosphate
-
-
-
-
r
beta-D-glucose-1-phosphate + 1,5-anhydro-D-glucitol
? + phosphate
-
-
-
?
beta-D-glucose-1-phosphate + 1,5-anhydro-D-glucitol
? + phosphate
-
-
-
?
D-glucose + beta-D-glucose 1-phosphate
2-alpha-D-glucosyl-D-glucose + phosphate
-
-
-
r
D-glucose + beta-D-glucose 1-phosphate
2-alpha-D-glucosyl-D-glucose + phosphate
-
-
kojioligosaccharides having only the alpha-1,2-linkage are synthesized and the average degree of polymerization of oligosaccharides increases with decreasing proportions of glucose
?
D-glucose + beta-D-glucose 1-phosphate
2-alpha-D-glucosyl-D-glucose + phosphate
-
-
[O-alpha-D-glucopyranosyl-(1-2)]m-O-[beta-D-fructofuranosyl-(2-1)]2-alpha-D-glucopyranoside with m = 1, 2 or 3
?
kojibiose + phosphate
beta-D-glucose-1-phosphate + D-glucose
-
-
-
-
?
kojibiose + phosphate
beta-D-glucose-1-phosphate + D-glucose
-
-
-
-
r
kojibiose + phosphate
beta-D-glucose-1-phosphate + D-glucose
-
-
-
-
r
kojitetraose + phosphate
beta-D-glucose 1-phosphate + kojitriose
-
-
-
-
r
kojitetraose + phosphate
beta-D-glucose 1-phosphate + kojitriose
-
-
-
-
r
kojitriose + phosphate
beta-D-glucose 1-phosphate + 2-alpha-D-glucosyl-D-glucose
-
-
-
-
r
kojitriose + phosphate
beta-D-glucose 1-phosphate + 2-alpha-D-glucosyl-D-glucose
-
-
-
-
r
additional information
?
-
-
the enzyme is absolutely specific for kojioligosaccharides in the phosphorolysis reaction, no activity with lactose, sucrose, trehalose, cellobiose, isomaltose, maltose, nigerose, and neotrehalose. For the synthesis reaction, the enzyme is specific for D-glucose, followed by kojibiose and kojitriose, overview
-
-
?
additional information
?
-
nigerose, maltose, and trehalose are minor substrates for the wild-type enzyme, substrate specificity of wild-type and mutant enzymes, overview
-
-
?
additional information
?
-
-
nigerose, maltose, and trehalose are minor substrates for the wild-type enzyme, substrate specificity of wild-type and mutant enzymes, overview
-
-
?
additional information
?
-
-
the enzyme is absolutely specific for kojioligosaccharides in the phosphorolysis reaction, no activity with lactose, sucrose, trehalose, cellobiose, isomaltose, maltose, nigerose, and neotrehalose. For the synthesis reaction, the enzyme is specific for D-glucose, followed by kojibiose and kojitriose, overview
-
-
?
additional information
?
-
no activity with trehalose or cellobiose. Activity with maltose or isomaltose es less than 0.001% of the activity with kojibiose
-
-
?
additional information
?
-
-
no activity with trehalose or cellobiose. Activity with maltose or isomaltose es less than 0.001% of the activity with kojibiose
-
-
?
additional information
?
-
no activity with trehalose or cellobiose. Activity with maltose or isomaltose es less than 0.001% of the activity with kojibiose
-
-
?
additional information
?
-
-
enzyme also catalyzes the chain-extending reaction of the side chain of 2-O-alpha-D-glucopyranosyl-CTS
-
-
?
additional information
?
-
-
no activity with ethanol, ethyleneglycol, 1-propanol, propyleneglycol, 1-butanol and 1,2-butanediol as acceptor
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
2-alpha-D-glucosyl-D-glucose i.e. kojibiose. The enzyme is involved in conversion of kojibiose into beta-D-glucose 6-phosphate, a substrate of the glycolytic pathway. Disaccharide utilization pathway via phosphorolysis in hyperthermophilic archaea
-
-
?
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
2-alpha-D-glucosyl-D-glucose i.e. kojibiose. The enzyme is involved in conversion of kojibiose into beta-D-glucose 6-phosphate, a substrate of the glycolytic pathway. Disaccharide utilization pathway via phosphorolysis in hyperthermophilic archaea
-
-
?
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
-
?
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
r
2-alpha-D-glucosyl-D-glucose + phosphate
D-glucose + beta-D-glucose 1-phosphate
-
-
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
DISEASE
TITLE OF PUBLICATION
LINK TO PUBMED
Erythema
Detection of keratinolytic proteinase in skin tissues from guinea pigs infected with Microsporum canis by an immunoperoxidase technique.
Infections
Detection of keratinolytic proteinase in skin tissues from guinea pigs infected with Microsporum canis by an immunoperoxidase technique.
Infections
Isolation and characterization of proteinase from Candida albicans: substrate specificity.
Tinea
Detection of keratinolytic proteinase in skin tissues from guinea pigs infected with Microsporum canis by an immunoperoxidase technique.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
9.3
2-alpha-D-glucosyl-D-glucose
-
pH 6.2, 45°C, recombinant loop 3 mutant 3 of maltose phosphorylase
16
2-alpha-D-glucosyl-D-glucose
-
pH 6.2, 45°C, recombinant loop 3 mutant 2 of maltose phosphorylase
84
2-alpha-D-glucosyl-D-glucose
-
pH 6.2, 45°C, recombinant loop 3 mutant 1 of maltose phosphorylase
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.097
2-alpha-D-glucosyl-D-glucose
-
pH 6.2, 45°C, recombinant loop 3 mutant 1 of maltose phosphorylase
0.12
2-alpha-D-glucosyl-D-glucose
-
pH 6.2, 45°C, recombinant loop 3 mutant 2 of maltose phosphorylase
2.4
2-alpha-D-glucosyl-D-glucose
-
pH 6.2, 45°C, recombinant loop 3 mutant 3 of maltose phosphorylase
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0011
2-alpha-D-glucosyl-D-glucose
-
pH 6.2, 45°C, recombinant loop 3 mutant 1 of maltose phosphorylase
0.0075
2-alpha-D-glucosyl-D-glucose
-
pH 6.2, 45°C, recombinant loop 3 mutant 2 of maltose phosphorylase
0.26
2-alpha-D-glucosyl-D-glucose
-
pH 6.2, 45°C, recombinant loop 3 mutant 3 of maltose phosphorylase
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
191.3
-
purified recombinant enzyme, pH 6.0, 50°C, synthesis
31.1
-
purified recombinant enzyme, pH 6.0, 50°C, phosphorolysis
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.5
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
brenda
strain ATCC35047, expression in Escherichia coli
-
-
brenda
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
physiological function
additional information
evolution
-
the enzyme belongs to the glycoside hydrolase family 65, GH65
evolution
-
the enzyme is a member of glycoside hydrolase family 65, GH65
evolution
-
the enzyme is a member of glycoside hydrolase family 65, GH65
evolution
the kojibiose phosphorylase belongs to the glycoside hydrolase (GH) family 65, that contains phosphorylases acting on maltose (Glc-alpha1,4-Glc), kojibiose (Glc-alpha1,2-Glc), trehalose (Glc-alpha1,alpha1,-Glc), and nigerose (Glc-alpha1,3-Glc). The loop 3 region comprising the active site of kojibiose phosphorylase is significantly longer than the active sites of other enzymes, and three residues around this loop, Trp391, Glu392, and Thr417, recognize kojibiose
evolution
-
the enzyme is a member of glycoside hydrolase family 65, GH65
-
evolution
-
the enzyme belongs to the glycoside hydrolase family 65, GH65
-
evolution
-
the enzyme is a member of glycoside hydrolase family 65, GH65
-
physiological function
the enzyme is involved in conversion of kojibiose into beta-D-glucose 6-phosphate, a substrate of the glycolytic pathway. Disaccharide utilization pathway via phosphorolysis in hyperthermophilic archaea
physiological function
-
the enzyme is involved in conversion of kojibiose into beta-D-glucose 6-phosphate, a substrate of the glycolytic pathway. Disaccharide utilization pathway via phosphorolysis in hyperthermophilic archaea
-
additional information
-
structure-function relationship analysis, homology modelling, overview. Substitution of maltose phosphorylase, EC 2.4.1.8, His413-Glu421, His413-Ile418 and His413-Glu415 from loop 3, that include His413 and Glu415 presumably recognising the alpha-anomeric O-1 group of the glucose moiety at subsite 1+, by corresponding to segments from Ser426-Ala431 in trehalose phosphorylase, EC 2.4.1.64, and Thr419-Phe427 in kojibiose phosphorylase, EC 2.4.1.230, thus confers the maltose phosphorylase with phosphorolytic activity towards trehalose and kojibiose, respectively. The loop 3 in GH65 disaccharide phosphorylase is a key determinant for specificity both in phosphorolysis and in regiospecific reverse phosphorolysis
additional information
-
structure-function relationship analysis, overview. The sequence Thr419-Phe427 is responsible for substrate specificity of the enzyme for kojibiose. The loop 3 in GH65 disaccharide phosphorylase is a key determinant for specificity both in phosphorolysis and in regiospecific reverse phosphorolysis
additional information
the loop 3 region comprises the active site of kojibiose, three residues around this loop, Trp391, Glu392, and Thr417, recognize kojibiose. Trp391 and Glu392, especially the latter, are required for the kojibiose activity, active site structure and substrate binding, overview. Comparison of substrate recognition by kojibiose phosphorylase CsKP from Caldicellulosiruptor saccharolyticus and maltose phosphorylase LbMP from Lactobacillus brevis, modeling, overview
additional information
-
the loop 3 region comprises the active site of kojibiose, three residues around this loop, Trp391, Glu392, and Thr417, recognize kojibiose. Trp391 and Glu392, especially the latter, are required for the kojibiose activity, active site structure and substrate binding, overview. Comparison of substrate recognition by kojibiose phosphorylase CsKP from Caldicellulosiruptor saccharolyticus and maltose phosphorylase LbMP from Lactobacillus brevis, modeling, overview
additional information
-
structure-function relationship analysis, homology modelling, overview. Substitution of maltose phosphorylase, EC 2.4.1.8, His413-Glu421, His413-Ile418 and His413-Glu415 from loop 3, that include His413 and Glu415 presumably recognising the alpha-anomeric O-1 group of the glucose moiety at subsite 1+, by corresponding to segments from Ser426-Ala431 in trehalose phosphorylase, EC 2.4.1.64, and Thr419-Phe427 in kojibiose phosphorylase, EC 2.4.1.230, thus confers the maltose phosphorylase with phosphorolytic activity towards trehalose and kojibiose, respectively. The loop 3 in GH65 disaccharide phosphorylase is a key determinant for specificity both in phosphorolysis and in regiospecific reverse phosphorolysis
-
additional information
-
structure-function relationship analysis, overview. The sequence Thr419-Phe427 is responsible for substrate specificity of the enzyme for kojibiose. The loop 3 in GH65 disaccharide phosphorylase is a key determinant for specificity both in phosphorolysis and in regiospecific reverse phosphorolysis
-
Show AA Sequence and Transmembrane Helices (603 entries)
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PDB
SCOP
CATH
UNIPROT
ORGANISM
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified enzyme in complex with glucose and phosphate and in complex with kojibiose and sulfate, Glc-PO4 crystals are obtained at 25°C by sitting drop vapor diffusion method, mixing 0.001 ml of 5.9 m/ml protein in 5 mM Tris/HCl, pH 7.5, with an equal volume of reservoir solution containing 5 mM D-glucose, 5 mM sodium phosphate, pH 8.5, 10% v/v 2-propanol, 10% w/v PEG 3350, and 0.1 M Tris/HCl, pH 8.5, the kojibiose-SO4 crystals are obtained at 25°C by hanging drop vapor diffusion method, mixing of 0.001 ml of 5.4 mg/ml protein solution in 5 mM Tris/HCl, pH 7.5, with an equal volume of reservoir solution containing 5 mM kojibiose, 0.2 M NaCl, 2.0 M (NH4)2SO4, and 0.1 M sodium cacodylate, pH 6.5, X-ray diffraction structure determination and analysis at 2.05-2.80 A resolution, structures PDB IDs 3WIR and 3WIQ, molecular replacement with the maltose phosphorylase structure from Lactobacillus brevis LbMP, PDB ID 1H54, as the search model
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
E392R
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
E392R/T417A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
E392R/T417I
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T417A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T417F
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T417I
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
W391M/E392V
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
W391M/E392V/T417A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
W391M/E392V/T417I
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D459N
-
increase in Km-values for kojibiose, beta-D-glucose 1-phosphate and glucose
additional information
additional information
-
substitution of maltose phosphorylase, EC 2.4.1.8, His413-Glu421, His413-Ile418 and His413-Glu415 from loop 3, that include His413 and Glu415 presumably recognising the alpha-anomeric O-1 group of the glucose moiety at subsite 1+, by corresponding to segments from Ser426-Ala431 in trehalose phosphorylase, EC 2.4.1.64, and Thr419-Phe427 in kojibiose phosphorylase, EC 2.4.1.230, thus confers the maltose phosphorylase with phosphorolytic activity towards trehalose and kojibiose, respectively
additional information
-
substitution of maltose phosphorylase, EC 2.4.1.8, His413-Glu421, His413-Ile418 and His413-Glu415 from loop 3, that include His413 and Glu415 presumably recognising the alpha-anomeric O-1 group of the glucose moiety at subsite 1+, by corresponding to segments from Ser426-Ala431 in trehalose phosphorylase, EC 2.4.1.64, and Thr419-Phe427 in kojibiose phosphorylase, EC 2.4.1.230, thus confers the maltose phosphorylase with phosphorolytic activity towards trehalose and kojibiose, respectively
-
additional information
-
chimeric phosphorylases are constructed of the kojibiose phosphorylase gene and the trehalose phosphorylase gene from Thermoanaerobacter brockii. Chimera V-III show not trehalose phosphorylase, but kojibiose phosphorylase activity, although only 125 amino acid residues in 785 residues of chimera VIII are from that of kojibiose phosphorylase. Chimera V-III have 1% of the specific activity of the wild-type kojibiose phosphorylase. The temperature profile and kinetic parameters of chimera V-III are remarkably changed as compared to those of the wild-type kojibiose phosphorylase. Chimera VIII protein exists as a monomer in solution, whereas wild-type kojibiose phosphorylase and trehalose phosphorylase are hexamer and dimer structures, respectively. The chimera acts on nigerose, sophorose and laminaribiose, in addition to kojibiose. Chimera VIII is also able to act on sophorose and laminaribiose in the absence of inorganic phosphate, and produces two trisaccharides, beta-D-glucosyl-(1,6)-laminaribiose and laminaritriose, from laminaribiose
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
3.5 - 10
-
recombinant enzyme, stable at, phosphorolysis
721869
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
70
75
70
-
50% residual activity, mutant D513N 110 min, wild-type 16 min
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
kojibiose phosphorylase is purified from a cell-free extract of Thermoanaerobacter brockii
recombinant His-tagged LaMP loop 3 mutants from Escherichia coli strain BL21(DE3) by affinity chromatography and dialysis
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, expression in Escherichia coli
-
expression of His-tagged LaMP loop 3 mutants in Escherichia coli strain BL21(DE3)
-
kojibiose phosphorylase gene and trehalose phosphorylase gene are intracellularly hyperexpressed under the control of the Bacillus amyloliquefaciens alpha-amylase promoter in Bacillus subtilis
-
recombinant expression of wild-type and mutant enzymes
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
synthesis
synthesis
-
synthesis of selaginose by kojibiose phosphorylase gene and trehalose phosphorylase gene intracellularly hyperexpressed under the control of the Bacillus amyloliquefaciens alpha-amylase promoter in Bacillus subtilis
synthesis
the phosphorylase can efficiently catalyze the reverse reaction with high specificity, and thus can be applied to the practical synthesis of alpha-glucosyl oligosaccharides
synthesis
-
synthesis of selaginose by kojibiose phosphorylase gene and trehalose phosphorylase gene intracellularly hyperexpressed under the control of the Bacillus amyloliquefaciens alpha-amylase promoter in Bacillus subtilis
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Chaen, H.; Yamamoto, T.; Nishimoto, T.; Nakada, T.; Fukuda, S.; Sugimoto, T.; Kurimoto, M.; Tsujisaka, Y.
Purification and characterization of a novel phosphorylase, kojibiose phosphorylase, from Thermoanaerobium brockii
J. Appl. Glycosci.
46
423-429
1999
Thermoanaerobacter brockii
-
brenda
Chaen, H.; Nishimoto, T.; Nakada, T.; Fukuda, S.; Kurimoto, M.; Tsujisaka, Y.
Enzymatic synthesis of kojioligosaccharides using kojibiose phosphorylase
J. Biosci. Bioeng.
92
177-182
2001
Thermoanaerobacter brockii
brenda
Okada, H.; Fukushi, E.; Onodera, S.; Nishimoto, T.; Kawabata, J.; Kikuchi, M.; Shiomi, N.
Synthesis and structural analysis of five novel oligosaccharides prepared by glucosyltransfer from beta-D-glucose 1-phosphate to isokestose and nystose using Thermoanaerobacter brockii kojibiose phosphorylase
Carbohydr. Res.
338
879-885
2003
Thermoanaerobacter brockii
brenda
Watanabe, H.; Higashiyama, T.; Aga, H.; Nishimoto, T.; Kubota, M.; Fukuda, S.; Kurimoto, M.; Tsujisaka, Y.
Enzymatic synthesis of a 2-O-alpha-D-glucopyranosyl cyclic tetrasaccharide by kojibiose phosphorylase
Carbohydr. Res.
340
449-454
2005
Thermoanaerobacter brockii
brenda
Yamamoto, T.; Mukai, K.; Yamashita, H.; Kubota, M.; Fukuda, S.; Kurimoto, M.; Tsujisaka, Y.
Enhancement of thermostability of kojibiose phosphorylase from Thermoanaerobacter brockii ATCC35047 by random mutagenesis
J. Biosci. Bioeng.
100
212-215
2005
Thermoanaerobacter brockii
brenda
Yamamoto, T.; Maruta, K.; Mukai, K.; Yamashita, H.; Nishimoto, T.; Kubota, M.; Fukuda, S.; Kurimoto, M.; Tsujisaka, Y.
Cloning and sequencing of kojibiose phosphorylase gene from Thermoanaerobacter brockii ATCC35047
J. Biosci. Bioeng.
98
99-106
2004
Thermoanaerobacter brockii, Thermoanaerobacter brockii ATCC 35047
brenda
Takahashi, N.; Okada, H.; Fukushi, E.; Onodera, S.; Nishimoto, T.; Kawabata, J.; Shiomi, N.
Structural analysis of six novel oligosaccharides synthesized by glucosyl transfer from beta-D-glucose 1-phosphate to raffinose and stachyose using Thermoanaerobacter brockii kojibiose phosphorylase
Tetrahedron
16
57-63
2005
Thermoanaerobacter brockii
-
brenda
Yamamoto, T.; Yamashita, H.; Mukai, K.; Watanabe, H.; Kubota, M.; Chaen, H.; Fukuda, S.
Construction and characterization of chimeric enzymes of kojibiose phosphorylase and trehalose phosphorylase from Thermoanaerobacter brockii
Carbohydr. Res.
341
2350-2359
2006
Thermoanaerobacter brockii
brenda
Yamamoto, T.; Mukai, K.; Maruta, K.; Watanabe, H.; Yamashita, H.; Nishimoto, T.; Kubota, M.; Chaen, H.; Fukuda, S.
Hyper expression of kojibiose phosphorylase gene and trehalose phosphorylase gene from Thermoanaerobacter brockii ATCC35047 in Bacillus subtilis and selaginose synthesis utilizing two phosphorylases
J. Biosci. Bioeng.
100
343-346
2005
Thermoanaerobacter brockii, Thermoanaerobacter brockii TCC35047
brenda
Yamamoto, T.; Watanabe, H.; Nishimoto, T.; Aga, H.; Kubota, M.; Chaen, H.; Fukuda, S.
Acceptor recognition of kojibiose phosphorylase from Thermoanaerobacter brockii: syntheses of glycosyl glycerol and myo-inositol
J. Biosci. Bioeng.
101
427-433
2006
Thermoanaerobacter brockii
brenda
Takahashi, N.; Fukushi, E.; Onodera, S.; Benkeblia, N.; Nishimoto, T.; Kawabata, J.; Shiomi, N.
Three novel oligosaccharides synthesized using Thermoanaerobacter brockii kojibiose phosphorylase
Chem. Cent. J.
1
18
2007
Thermoanaerobacter brockii (Q8L163), Thermoanaerobacter brockii
brenda
Takahashi, N.; Fukushi, E.; Onodera, S.; Nishimoto, T.; Kawabata, J.; Shiomi, N.
Isolation and identification of novel tri- and tetra-saccharides synthesized by Thermoanaerobacter brockii kojibiose phosphorylase
J. Appl. Glycosci.
54
195-200
2007
Thermoanaerobacter brockii (Q8L163)
-
brenda
Nakai, H.; Petersen, B.O.; Westphal, Y.; Dilokpimol, A.; Abou Hachem, M.; Duus, J.?.; Schols, H.A.; Svensson, B.
Rational engineering of Lactobacillus acidophilus NCFM maltose phosphorylase into either trehalose or kojibiose dual specificity phosphorylase
Protein Eng. Des. Sel.
23
781-787
2010
Thermoanaerobacter brockii, Lactobacillus acidophilus, Thermoanaerobacter brockii ATCC 35047
brenda
Yamamoto, T.; Nishio-Kosaka, M.; Izawa, S.; Aga, H.; Nishimoto, T.; Chaen, H.; Fukuda, S.
Enzymatic properties of recombinant kojibiose phosphorylase from Caldicellulosiruptor saccharolyticus ATCC43494
Biosci. Biotechnol. Biochem.
75
1208-1210
2011
Caldicellulosiruptor saccharolyticus, Caldicellulosiruptor saccharolyticus ATCC 43494
brenda
Jung, J.H.; Seo, D.H.; Holden, J.F.; Park, C.S.
Identification and characterization of an archaeal kojibiose catabolic pathway in the hyperthermophilic Pyrococcus sp. strain ST04
J. Bacteriol.
196
1122-1131
2014
Pyrococcus sp. (I3RFR4), Pyrococcus sp., Pyrococcus sp. ST04 (I3RFR4)
brenda
Okada, S.; Yamamoto, T.; Watanabe, H.; Nishimoto, T.; Chaen, H.; Fukuda, S.; Wakagi, T.; Fushinobu, S.
Structural and mutational analysis of substrate recognition in kojibiose phosphorylase
FEBS J.
281
778-786
2014
Caldicellulosiruptor saccharolyticus (A4XGP2), Caldicellulosiruptor saccharolyticus
brenda
Mukherjee, K.; Narindoshvili, T.; Raushel, F.M.
Discovery of a kojibiose phosphorylase in Escherichia coli K-12
Biochemistry
57
2857-2867
2018
Escherichia coli (P77154), Escherichia coli MG1655 (P77154)
brenda
Select items on the left to see more content.
EXTERNAL LINKS (specific for EC-Number 2.4.1.230)
html completed
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
About BRENDA
Social media
Help
Survey
Download
Contribution
Legal
Curated at
Member of
