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Information on EC 2.4.1.82 - galactinol-sucrose galactosyltransferase and Organism(s) Arabidopsis thaliana and UniProt Accession Q8RX87
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EC Tree
2.4.1.82 galactinol-sucrose galactosyltransferaseIUBMB Comments
4-Nitrophenyl alpha-D-galactopyranoside can also act as donor. The enzyme also catalyses an exchange reaction between raffinose and sucrose (cf. EC 2.4.1.123, inositol 3-alpha-galactosyltransferase).
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Word Map
- 2.4.1.82
- oligosaccharide
- stachyose
- verbascose
-
monogastric
- agriculture
-
cold-responsive
-
antinutritional
- synthesis
The taxonomic range for the selected organisms is: Arabidopsis thaliana
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Synonyms
raffinose synthase, zmrafs, raffinose synthase 2, galactinol-sucrose galactosyltransferase, raffinose synthase 5, raffinose synthase 4, raffinose synthase 3, raffinose synthase 6, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
galactosyltransferase, galactinol-sucrose
-
-
-
-
raffinose synthase
RafS
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hexosyl group transfer
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
alpha-D-galactosyl-(1->3)-1D-myo-inositol:sucrose 6-alpha-D-galactosyltransferase
4-Nitrophenyl alpha-D-galactopyranoside can also act as donor. The enzyme also catalyses an exchange reaction between raffinose and sucrose (cf. EC 2.4.1.123, inositol 3-alpha-galactosyltransferase).
CAS REGISTRY NUMBER
COMMENTARY
62213-45-0
-
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
galactinol + sucrose
myo-inositol + raffinose
-
-
-
ir
galactinol + sucrose
myo-inositol + raffinose
-
-
-
ir
myo-inositol + raffinose
1-alpha-D-galactosyl-1D-myo-inositol + sucrose
-
-
-
-
r
alpha-D-galactosyl-(1->3)-1D-myo-inositol + sucrose
myo-inositol + raffinose
-
-
-
?
alpha-D-galactosyl-(1->3)-1D-myo-inositol + sucrose
myo-inositol + raffinose
-
-
-
?
additional information
?
-
the multifunctional enzyme is a raffinose and high affinity stachyose synthase, EC 2.4.1.82 and EC 2.4.1.67, as well as stachyose and galactinol specific galactosylhydrolase
-
-
?
additional information
?
-
-
the multifunctional enzyme is a raffinose and high affinity stachyose synthase, EC 2.4.1.82 and EC 2.4.1.67, as well as stachyose and galactinol specific galactosylhydrolase
-
-
?
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
alpha-D-galactosyl-(1->3)-1D-myo-inositol + sucrose
myo-inositol + raffinose
-
-
-
?
alpha-D-galactosyl-(1->3)-1D-myo-inositol + sucrose
myo-inositol + raffinose
-
-
-
?
additional information
?
-
the multifunctional enzyme is a raffinose and high affinity stachyose synthase, EC 2.4.1.82 and EC 2.4.1.67, as well as stachyose and galactinol specific galactosylhydrolase
-
-
?
additional information
?
-
-
the multifunctional enzyme is a raffinose and high affinity stachyose synthase, EC 2.4.1.82 and EC 2.4.1.67, as well as stachyose and galactinol specific galactosylhydrolase
-
-
?
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
heat shock transcription factor A2
in HsfA2 overexpressing plants transcription levels of RS2 increases 5fold
-
additional information
-
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS6 4fold after 3 hours and 6fold after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS6 4fold after 3 hours and 6fold after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS6 4fold after 3 hours and 6fold after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS6 4fold after 3 hours and 6fold after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS6 4fold after 3 hours and 6fold after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS6 4fold after 3 hours and 6fold after 6 hours
-
additional information
-
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS2 5fold after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS2 5fold after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS2 5fold after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS2 5fold after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS2 5fold after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS2 5fold after 6 hours
-
additional information
-
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS4 3fold after 3 and 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS4 3fold after 3 and 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS4 3fold after 3 and 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS4 3fold after 3 and 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS4 3fold after 3 and 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS4 3fold after 3 and 6 hours
-
additional information
-
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS5 twice after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS5 twice after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS5 twice after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS5 twice after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS5 twice after 6 hours
-
additional information
50 microM methylviologen, an enhancer of oxidative stress, increases transcription levels of RS5 twice after 6 hours
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.17
alpha-D-galactosyl-(1->3)-1D-myo-inositol
pH 7.0, 25°C, recombinant His-tagged enzyme
additional information
additional information
Michaelis-Menten kinetics
-
additional information
additional information
-
Michaelis-Menten kinetics
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.5
-
assay at, RafS enzyme activity shows neutral pH optima, corresponding to the pH of the cytosol
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
heat shock transcription factor A2 HsfA2 overexpressing plants
UniProt
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
raffinose synthase 6 plays a role in protection from oxidative stress
malfunction
metabolism
physiological function
malfunction
leaves of RFS5 mutant plants fail to accumulate any raffinose under diverse abiotic stresses including water-deficit, high salinity, heat shock, and methyl viologen-induced oxidative stress. Correlated to the lack of raffinose under these abiotic stress conditions, both mutant plants lack the typical stress-induced raffinose synthase activity increase observed in the leaves of wild-type plants
malfunction
metabolite phenotype of DELTAAtRS4 mutant seeds, overview
metabolism
biosynthesis of RFOs proceeds by stepwise transfer of galactosyl units. The second step involves raffinose synthase, AtRS5, EC 2.4.1.82, which transfers the galactosyl unit from galactinol to the C6 position of the glucose unit in sucrose forming an alpha-1,6-galactosidic linkage to yield the trisaccharide raffinose. In a third step, stachyose synthase, AtRS4, EC 2.4.1.67, transfers the galactosyl moiety from galactinol to the C6 position of the galactose unit in raffinose to yield the tetrasaccharide stachyose
metabolism
the enzyme catalyzes the second step in raffinose biosynthesis
physiological function
raffinose synthase 2 plays a role in heat shock reaction and protection from oxidative stress
physiological function
raffinose synthase 4 plays a role in protection from oxidative stress
physiological function
raffinose synthase 5 plays a role in protection from oxidative stress
physiological function
AtRS4 is the only stachyose synthase in the genome of Arabidopsis thaliana. It represents a key regulation mechanism in the raffinose family oligosaccharide physiology of Arabidosis thaliana due to its multifunctional enzyme activity. AtRS4 is possibly the second seed-specific raffinose synthase beside AtRS5, which is responsible for raffinose accumulation under abiotic stress
physiological function
the enzyme is responsible for low temperature-induced raffinose accumulation in Arabidopsis thaliana leaves
physiological function
in seeds of the galactinol synthase GS1/GS2 and raffinose synthase RFS5 mutant the timing of desiccation tolerance acquisition is delayed as compared to wild type. Seeds from GS1/GS2 overexpressing plants with high levels of galactinol, raffinose, and stachyose, and RS5 overexpressing plants possess more raffinose and stachyose but less galactinol compared to wild type. These lines show greater germination percentage and shorter time to 50% germination after desiccation treatment at 11 and 15 days after flower. The role of raffinose family oligosaccharides is time limited and mainly affects the middle stage of seed development by enhancing seed viability and the ratio of GSH to GSSH in cells, but there is no significant difference in desiccation tolerance of mature seeds
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). RFS6_ARATH
749
0
83116
Swiss-Prot
other Location (Reliability: 4)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
100000
x * 98010, about, sequence calculation, x * 100000, recombinant His-tagged enzyme, SDS-PAGE
98010
x * 98010, about, sequence calculation, x * 100000, recombinant His-tagged enzyme, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
x * 98010, about, sequence calculation, x * 100000, recombinant His-tagged enzyme, SDS-PAGE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
construction of two independent loss-of-function mutants for gene RFS5, i.e. rs 5-1 and rs 5-2. Leaves of mutant plants fail to accumulate any raffinose under diverse abiotic stresses including water-deficit, high salinity, heat shock, and methyl viologen-induced oxidative stress. Correlated to the lack of raffinose under these abiotic stress conditions, both mutant plants lack the typical stress-induced raffinose synthase activity increase observed in the leaves of wild-type plants. The seeds of both mutant plants still contain raffinose Phenotypes, overview
additional information
isolation of a T-DNA insertional mutant in the AtRS4 gene, only semi-quantitative PCR from wild-type siliques shows a specific transcriptional AtRS4PCR product. Metabolite measurements in seeds of DELTAAtRS4 mutant plants reveal a total loss of stachyose in DELTAAtRS4 mutant seeds
additional information
-
isolation of a T-DNA insertional mutant in the AtRS4 gene, only semi-quantitative PCR from wild-type siliques shows a specific transcriptional AtRS4PCR product. Metabolite measurements in seeds of DELTAAtRS4 mutant plants reveal a total loss of stachyose in DELTAAtRS4 mutant seeds
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
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene RFS4, cloning from seeds and functional recombinant expression of His-tagged enzyme in Escherichia coli
gene RFS5, functional recombinant expression in Escherichia coli strain BL21 codon plus
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
agriculture
in seeds of the galactinol synthase GS1/GS2 and raffinose synthase RFS5 mutant the timing of desiccation tolerance acquisition is delayed as compared to wild type. Seeds from GS1/GS2 overexpressing plants with high levels of galactinol, raffinose, and stachyose, and RS5 overexpressing plants possess more raffinose and stachyose but less galactinol compared to wild type. These lines show greater germination percentage and shorter time to 50% germination after desiccation treatment at 11 and 15 days after flower. The role of raffinose family oligosaccharides is time limited and mainly affects the middle stage of seed development by enhancing seed viability and the ratio of GSH to GSSH in cells, but there is no significant difference in desiccation tolerance of mature seeds
synthesis
use of an in vitro multienzyme system using sucrose synthase (SUS), UDP-glucose 4-epimerase (GalE), galactinol synthase (GS), raffinose synthase (RS), and stachyose synthase (STS) and intermedia UDP and inositol, which can be recycled, to synthesize raffinose,. The reaction system produces 11.1mM raffinose using purified enzymes under optimal reaction conditions and substrate concentrations. Optimization further increases raffinose production to 86.6 mM and enables the synthesis of 61.1 mM stachyose. The UDP turnover number reaches 337. Inositol in the reaction system is recycled five times, and 255.8 mM raffinose (128.9 g/liter) is obtained
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Nishizawa, A.; Yabuta, Y.; Shigeoka, S.
Galactinol and raffinose constitute a novel function to protect plants from oxidative damage
Plant Physiol.
147
1251-1263
2008
Arabidopsis thaliana, Arabidopsis thaliana (Q84VX0), Arabidopsis thaliana (Q8RX87), Arabidopsis thaliana (Q94A08), Arabidopsis thaliana (Q9FND9), Arabidopsis thaliana (Q9SYJ4)
Schneider, T.; Keller, F.
Raffinose in chloroplasts is synthesized in the cytosol and transported across the chloroplast envelope
Plant Cell Physiol.
50
2174-2182
2009
Ajuga reptans, Arabidopsis thaliana, Spinacia oleracea
Egert, A.; Keller, F.; Peters, S.
Abiotic stress-induced accumulation of raffinose in Arabidopsis leaves is mediated by a single raffinose synthase (RS5, At5g40390)
BMC Plant Biol.
13
218
2013
Arabidopsis thaliana (Q9FND9)
Gangl, R.; Behmueller, R.; Tenhaken, R.
Molecular cloning of AtRS4, a seed specific multifunctional RFO synthase/galactosylhydrolase in Arabidopsis thaliana
Front. Plant Sci.
6
789
2015
Arabidopsis thaliana (Q9SYJ4), Arabidopsis thaliana, Arabidopsis thaliana Col-0 (Q9SYJ4)
Tian, C.; Yang, J.; Zeng, Y.; Zhang, T.; Zhou, Y.; Men, Y.; You, C.; Zhu, Y.; Sun, Y.
Biosynthesis of raffinose and stachyose from sucrose via an in vitro multienzyme system
Appl. Environ. Microbiol.
85
e02306
2019
Arabidopsis thaliana (Q9SYJ4)
EXTERNAL LINKS (specific for EC-Number 2.4.1.82)
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