Information on EC 2.4.1.67 - galactinol-raffinose galactosyltransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.4.1.67
-
RECOMMENDED NAME
GeneOntology No.
galactinol-raffinose galactosyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
alpha-D-galactosyl-(1->3)-1D-myo-inositol + raffinose = myo-inositol + stachyose
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexosyl group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Galactose metabolism
-
-
stachyose biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
alpha-D-galactosyl-(1->3)-1D-myo-inositol:raffinose galactosyltransferase
This enzyme also catalyses galactosyl transfer from stachyose to raffinose (shown by labelling) [4]. For synthesis of the substrate, see EC 2.4.1.123, inositol 3-alpha-galactosyltransferase. See also EC 2.4.1.82, galactinol---sucrose galactosyltransferase.
CAS REGISTRY NUMBER
COMMENTARY hide
37277-70-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
frost.hardy species
-
-
Manually annotated by BRENDA team
ecotype Columbia; gene RFS4, isozyme RS4
-
-
Manually annotated by BRENDA team
gene RFS4, isozyme RS4
-
-
Manually annotated by BRENDA team
cv. inodorus
-
-
Manually annotated by BRENDA team
var. melopepo f. torticolis Bailey
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
no activity in Spodoptera frugiperda
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
cv. Wunder von Kelvedon
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-alpha-D-galactosyl-myo-inositol + D-ononitol
myo-inositol + alpha-D-galactosyl-D-ononitol
show the reaction diagram
1-alpha-D-galactosyl-myo-inositol + D-pinitol
myo-inositol + galactopinitol A
show the reaction diagram
1-alpha-D-galactosyl-myo-inositol + galactopinitol A
myo-inositol + ciceritol
show the reaction diagram
-
-
i.e. O-alpha-D-galactopyranosyl-(1,6)-O-alpha-D-galactopyranosyl-(1,2)-4-O-methyl-D-chiro-inositol
?
1-alpha-D-galactosyl-myo-inositol + H2O
myo-inositol + D-galactose
show the reaction diagram
1-alpha-D-galactosyl-myo-inositol + melibiose
myo-inositol + alpha-D-galactosyl-6-O-D-galactosyl-alpha-D-glucose
show the reaction diagram
1-alpha-D-galactosyl-myo-inositol + myo-inositol
myo-inositol + 1-alpha-D-galactosyl-myo-inositol
show the reaction diagram
1-alpha-D-galactosyl-myo-inositol + raffinose
myo-inositol + stachyose
show the reaction diagram
1-alpha-D-galactosyl-myo-inositol + sequoyitol
myo-inositol + alpha-D-galactosyl-sequoyitol
show the reaction diagram
1-alpha-D-galactosyl-myo-inositol + stachyose
myo-inositol + verbascose
show the reaction diagram
12.1% activity compared to raffinose
-
?
1-alpha-D-galactosyl-myo-inositol + verbascose
myo-inositol + ajugose
show the reaction diagram
-
-
?
alpha-D-galactosyl-(1->3)-1D-myo-inositol + raffinose
myo-inositol + stachyose
show the reaction diagram
alpha-D-galactosyl-D-ononitol + raffinose
D-ononitol + stachyose
show the reaction diagram
alpha-D-galactosyl-D-ononitol + stachyose
D-ononitol + verbascose
show the reaction diagram
9.9% activity compared to 1-alpha-D-galactosyl-myo-inositol
-
?
galactinol + raffinose
?
show the reaction diagram
-
assay at pH 7.0, 25°C, 90 min, reaction terminated by addition of NaOH
-
-
?
galactinol + raffinose
myo-inositol + stachyose
show the reaction diagram
-
assay at pH 7.0, 25°C, 90 min, reaction terminated by addition of NaOH
-
-
?
p-nitrophenyl-alpha-D-galactopyranoside + raffinose
p-nitrophenol + stachyose
show the reaction diagram
-
about 10% activity compared to 1-alpha-D-galactosyl-myo-inositol
-
r
stachyose + stachyose
raffinose + verbascose
show the reaction diagram
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
1-alpha-D-galactosyl-myo-inositol + D-ononitol
myo-inositol + alpha-D-galactosyl-D-ononitol
show the reaction diagram
-
biosynthesis of galactosylononitol
-
-
-
1-alpha-D-galactosyl-myo-inositol + raffinose
myo-inositol + stachyose
show the reaction diagram
alpha-D-galactosyl-(1->3)-1D-myo-inositol + raffinose
myo-inositol + stachyose
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dithiothreitol
-
activation
EDTA
-
slight stimulation
additional information
-
K-phosphate buffer is preferred
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
abscisic acid
-
in concentration of 250 micromol/l
Ag+
-
weak
Ca2+
-
weak
D-ononitol
-
competitive inhibition when the enzyme is incubated with both raffinose and D-ononitol simultaneously
melibiose
-
i.e. 6-O-alpha-D-galactosyl-alpha-D-glucose, non-competitive to raffinose, weak
Mg2+
-
weak
myo-inositol
-
strong, competitive
raffinose
-
competitive inhibition when the enzyme is incubated with both raffinose and D-ononitol simultaneously
Tris buffer
-
0.1 M
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
abscisic acid
-
in concentration of 50 and 150 micromol/l
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.33 - 7.7
1-alpha-D-galactosyl-myo-inositol
31.3
alpha-D-galactosyl-D-ononitol
-
-
12.4
D-pinitol
-
-
5.2
melibiose
-
+ 1-alpha-D-galactosyl-myo-inositol
5.5
myo-inositol
-
with 10 mM 1-alpha-D-galactosyl-myo-inositol in the exchange reaction
0.2592 - 38.6
raffinose
additional information
additional information
-
Michaelis-Menten kinetics
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
18.7
D-ononitol
-
competitive against raffinose
2
myo-inositol
-
-
17.9
raffinose
-
competitive against D-ononitol
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00011
-
minor-vein-enriched fraction
0.0004167
-
value about, after 6 days of cold treatment and 250 micromol/l exogenous abscisic acid
0.000833
-
leaf
0.00167
-
value about, cold-sensitve cultivar, Guonong No. 41, grown under low temperature for 9 days; value about, cold-tolerant cultivar, Guonong No. 25, grown under normal temperature
0.0033
-
value about, cold-sensitve cultivar, Guonong No. 41, grown under low temperature for 3 days; value about, cold-tolerant cultivar, Guonong No. 25, grown under low temperature for 3 days
0.004167
-
value about, after 6 days of cold treatment and 50 micromol/l exogenous abscisic acid
0.00458
-
value about, after 6 days of cold treatment and 150 micromol/l exogenous abscisic acid
0.0067
0.0083
-
value about, mesocarp tissue, 8 days after anthesis
0.01
-
value about, leaf, day of anthesis; value about, mesocarp tissue, 4 days after anthesis
0.01167
-
value about, mesocarp tissue, day of anthesis
0.0125
-
value about, leaf, 4 days after anthesis
0.013
-
value about, mesocarp tissue, 2 days before anthesis
0.0167
-
value about, leaf, 8 days after anthesis
0.0185
-
partially purified enzyme
0.02
recombinant enzyme in Sf21 cells, substrates galactosylononitol and raffinose
0.02083
-
value about, leaf, 20 days after anthesis
0.025
-
value about, leaf, 16 days after anthesis
0.02917
-
value about, leaf, 12 days after anthesis
0.04167
-
value about, mesocarp tissue, 20 days after anthesis
0.045
recombinant enzyme in Sf21 cells, substrates 1-alpha-D-galactosyl-myo-inositol and D-ononitol
0.05
recombinant enzyme in Sf21 cells, substrates 1-alpha-D-galactosyl-myo-inositol and raffinose
0.28
-
purified recombinant enzyme, pH 7.0, 25°C
0.5
purified enzyme
0.55
-
purified enzyme
0.67
-
purified enzyme
1.3
-
purified enzyme, substrates: 1-alpha-D-galactosyl-myo-inositol + D-ononitol
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 7
-
-
6.5 - 6.9
-
plateau
6.5 - 7
-
-
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.4 - 7.6
-
about half-maximal activity at pH 5.4 and pH 7.6
6 - 8
-
tested range, maximal activity at pH 6.0, 65.8% of maximal activity at pH 8.0, 89.5% at pH 7.0
7 - 8
-
89.5% and 65.8% of activity at pH 7.0 and pH 8.0, respectively
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 35
-
activity range, inactive at 0°C and 40°C; at 5°C, 22.2% of enzyme activity is sustained, whereas 98.3% of the activity is lost at 40°C caused by protein inactivation
6 - 25
-
Q10 of 2.2
23 - 35
-
half-maximal activity at 23°C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
activity is highly increased during cold-adaptation
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
mesophyll
-
Manually annotated by BRENDA team
additional information
-
extravacuolar
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
88600
-
1 * 88600, SDS-PAGE
90100
-
1 * 90100, SDS-PAGE
94900
DNA and amino acid sequence determination
95000
1 * 95000, about, SDS-PAGE
98000
1 * 98000, recombinant enzyme, SDS-PAGE
98001
-
1 * 98001, calculated from amino acid sequence
98010
-
x * 98010, about, sequence calculation, x * 100000, recombinant His-tagged enzyme, SDS-PAGE
99000
-
gel filtration
100000
-
gel filtration
110000
-
gel filtration
additional information
-
enzyme forms a 283 kDa aggregate during gel filtration step
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
unstable at low pH-values
638265
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Mg2+, bovine serum albumin, Triton, Tween 20, and DTT stabilize the enzyme, the latter is used in the enzyme assay
-
utilization of Na-phosphate and MES buffer (pH 7.0) leads to 86.5% and 82.5% of enzyme activity, respectively, compared with 100% enzyme activity in K-phosphate buffer at the same pH
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-196°C, stored in liquid N2, stable for at least 4 months
-
-20°C, loss of 85% activity within 4 months
-
4°C, 0.1 M sodium phosphate buffer, 20 mM 2-mercaptoethanol, up to 45 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
desalted
-
nickel affinity column chromatography; recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography
-
partial
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloned from maturing seeds, DNA sequence determination and analysis, functional expression in Spodoptera frugiperda Sf9 insect cells via baculovirus infection
DNA and amino acid determination, functional expression in Spodoptera frugiperda Sf21 insect cells via baculovirus infection
expressed in Arabidopsis thaliana
-
expressed in Escherichia coli; gene RFS4, cloning from seeds and functional recombinant expression of His-tagged enzyme in Escherichia coli
-
transformation in Agrobacterium tumefaciens, infiltratet in Arabidopsis thaliana
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
enhanced expression when grown on agar plates supplemented with 4% sucrose under high light growth conditions
enzyme expression decreases strongly during 20 days of dehydration but increases again following 24 h rehydration
-
highest expression levels in mature leaves and in cotyledons, followed by young leaves; no expression detected in stem, root, flower and fruit
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information