Information on EC 2.4.1.14 - sucrose-phosphate synthase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
2.4.1.14
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RECOMMENDED NAME
GeneOntology No.
sucrose-phosphate synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
UDP-glucose + D-fructose 6-phosphate = UDP + sucrose 6F-phosphate
show the reaction diagram
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-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexosyl group transfer
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-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Metabolic pathways
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Starch and sucrose metabolism
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sucrose biosynthesis I (from photosynthesis)
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sucrose biosynthesis II
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sucrose biosynthesis III
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SYSTEMATIC NAME
IUBMB Comments
UDP-glucose:D-fructose-6-phosphate 2-alpha-D-glucosyltransferase
Requires Mg2+ or Mn2+ for maximal activity [2]. The enzyme from Synechocystis sp. strain PCC 6803 is not specific for UDP-glucose as it can use ADP-glucose and, to a lesser extent, GDP-glucose as substrates [2]. The enzyme from rice leaves is activated by glucose 6-phosphate but that from cyanobacterial species is not [2]. While the reaction catalysed by this enzyme is reversible, the enzyme usually works in concert with EC 3.1.3.24, sucrose-phosphate phosphatase, to form sucrose, making the above reaction essentially irreversible [3]. The F in sucrose 6F-phosphate is used to indicate that the fructose residue of sucrose carries the substituent.
CAS REGISTRY NUMBER
COMMENTARY hide
9030-06-2
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain 7119
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-
Manually annotated by BRENDA team
strain 7119
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-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
rape
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-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
cotton
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-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Oncidium Goldiana
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
several cultivars., e.g. ROC20 and RB72-454
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-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
Gandoger
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-
Manually annotated by BRENDA team
Gandoger
-
-
Manually annotated by BRENDA team
Synechococcus marinus
-
-
-
Manually annotated by BRENDA team
ladino clover
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
cultered mesophyll cells
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-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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a comparison of the sequences of the wheat SPSII orthologues present in the diploid progenitors Triticum monococcum, Triticum urartu, Triticum speltoides, Aegilops tauschii, and Triticum speltoides, as well as in the more distantly related species Hordeum vulgare, Oryza sativa, Sorghum and purple false brome, Brachypodium distachyon, demonstrates that intronic sequence is less well conserved than exonic. Comparative sequence and phylogenetic analysis of SPSII gene shows that false purple brome is more similar to Triticeae than to Oryza sativa
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ADP-glucose + D-fructose 6-phosphate
ADP + sucrose 6-phosphate
show the reaction diagram
GDP-glucose + D-fructose 6-phosphate
GDP + sucrose 6-phosphate
show the reaction diagram
UDP-alpha-D-glucose + D-fructose 6-phosphate
UDP + sucrose 6-phosphate
show the reaction diagram
UDP-glucose + D-fructose 6-phosphate
UDP + sucrose 6F-phosphate
show the reaction diagram
additional information
?
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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
UDP-alpha-D-glucose + D-fructose 6-phosphate
UDP + sucrose 6-phosphate
show the reaction diagram
UDP-glucose + D-fructose 6-phosphate
UDP + sucrose 6F-phosphate
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
no effect
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5-azido-UDP-glucose
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nearly complete inhibition at 1 mM
citrate
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activity can by restored with Mg2+
cycloheximide
D-fructose 6-phosphate
-
inhibitory at high concentrations
delta-gluconolactone
diphosphate
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slight inhibition
fluoride
fructose-1,6-bisphosphate
-
-
ITP
-
reversible by addition of Mg2+
NaCl
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10% of activity remaining above 0.2 M
NaF
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strong inhibition at 20 mM
okadaic acid
-
-
p-chloromercuribenzoate
phosphate
phosphoenolpyruvate
-
slight inhibition
Sucrose
sucrose-6-phosphate
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sulfate
Tris-HCl buffer
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slight inhibition
UDP-glucose
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-
vanadate
-
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,5-Anhydroglucitol 6-phosphate
benzyladenine
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the phytohormone increases the specific activity of the enzyme practically during the entire growing period, except for the youngest leaves
D-fructose-1,6-diphosphate
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D-fructose-1-phosphate
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slight activation
D-glucose
D-glucose 6-phosphate
157-165% activation, the activation effect is reduced in plants infected with an N-fixation deficient Sinorhizobium meliloti strain
D-glucose-1-phosphate
D-Glucose-6-phosphate
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the enzyme is activated in light, with an increased affinity for its substrates and the activator glucose-6-phosphate, reduces sensitivity to inhibition by phosphate, but no change in maximal catalytic activity
fluoride
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gibberellic acid
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the phytohormone increases the specific activity of the enzyme practically during the entire growing period, except for the youngest leaves
glucosamine
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glucose-6-phosphate
IAA
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the phytohormone increases specific activity of the enzyme practically during the entire growing period, except for the youngest leaves
lactose
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maltose
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mannose
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pretreatment lowers sensitivity to phosphate inhibition
phosphate
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stimulating at low concentration particularly at high fructose-6-phosphate concentrations
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2 - 2.7
ADP-glucose
0.2 - 9.3
D-fructose 6-phosphate
1.5 - 2.4
GDP-glucose
1.3 - 77
UDP-glucose
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.8
fructose-1,6-bisphosphate
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-
1.75 - 11
phosphate
50
Sucrose
-
-
0.4
sucrose-6-phosphate
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0.7 - 3.6
UDP
9.4
UDP-glucose
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0025
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Vmax in vivo
0.003
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value about, mesocarp tissue, day of anthesis
0.0067
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value about, mesocarp tissue, 2 days before anthesis
0.01
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value about, mesocarp tissue, 20 days after anthesis
0.01167
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value about, leaf, 20 days after anthesis
0.013
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value about, mesocarp tissue, 16 days after anthesis; value about, mesocarp tissue, 4 days after anthesis
0.015
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value about, leaf, 2 days before anthesis
0.0167
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value about, leaf, 16 days after anthesis; value about, leaf, day of anthesis; value about, mesocarp tissue, 8 days and 12 days after anthesis
0.023
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value about, leaf, 12 days after anthesis
0.025
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value about, leaf, 4 days after anthesis
0.03
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value about, leaf, 8 days after anthesis
0.0316
wild-type leaf isozyme from plants infected with an N-fixation deficient Sinorhizobium meliloti strain
0.0322
wild-type leaf isozyme from plants infected with a wild-type Sinorhizobium meliloti strain
0.04
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purified SPS-I
0.0767
wild-type nodule isozyme from plants infected with an N-fixation deficient Sinorhizobium meliloti strain
0.0861
wild-type nodule isozyme from plants infected with a wild-type Sinorhizobium meliloti strain
0.25
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purified SPS-II
0.68
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purified enzyme
1.1
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purified enzyme
2.9
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purified enzyme
4.22
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purified enzyme
25
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purified enzyme
28
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purified enzyme, specific activity in crude extract is extremely low compared to spinach
57
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purified enzyme
79.5
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purified enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
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broad optimum
6.5
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broad optimum
6.5 - 7.5
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broad optimum
7.4
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assay at
8.5
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recombinant enzyme expressed in Escherichia coli
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 9
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 55
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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maximal activity of SPS in illumination grown calli appears on the 14th day after the culture is transferred into a new medium. Very low activity in dark grown cells
Manually annotated by BRENDA team
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although the level of SPS mRNA and protein is lower in embryos than in leaf, enzymatic activity is higher
Manually annotated by BRENDA team
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the level of SPS transcript is 10fold lower in endosperm than in leaf but the level of SPS protein is comparable and the activity is 2fold higher
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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low activity
Manually annotated by BRENDA team
high expression level of SPSA, nodules of plants inoculated with Sinorhizobium meliloti
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Halothermothrix orenii (strain H 168 / OCM 544 / DSM 9562)
Halothermothrix orenii (strain H 168 / OCM 544 / DSM 9562)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
46000
-
gel filtration
52000
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SDS-PAGE, minor band, degradation product
116000
117000
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calculated from DNA sequence
117600
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calculated from DNA sequence
118500
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calculated from DNA sequence
120000
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SDS-PAGE
130000 - 140000
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SDS-PAGE
138000
240000
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native PAGE
253000
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sucrose density gradient centrifugation
270000 - 280000
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gel filtration
270000
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gel filtration after Mono Q affinity column
380000 - 390000
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gel filtration, glycerol density gradient centrifugation
413000
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gel filtration of crude leaf extracts
420000
-
gel filtration
443000
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gel filtration
456000
-
gel filtration
460000
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gel filtration
480000
540000
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native PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 82000, SDS-PAGE
dimer
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2 * 120000, sucrose density gradient centrifugation, SDS-PAGE
monomer
tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging-drop vapor diffusion technique at 25°C. Crystal structure of SPS and its complexes with the substrate D-fructose 6-phosphate and the product D-sucrose-6'-phosphate. SPS has two distinct Rossmann-fold domains with a large substrate binding cleft at the interdomain interface. Structures of two complexes show that both the substrate D-fructose 6-phosphate and the product D-fructose 6'-phosphate bind to the A-domain of SPS. Halothermothrix orenii may represent a valid model for the catalytic domain of plant SPSs and thus may provide useful insight into the reaction mechanism of the plant enzyme
spsA protein crystallized in the monocyclic space group C2, with unit-cell parameters a = 154.2, b = 47.9, c = 103.16°, using hanging-drop vapour-diffusion method. Crystals diffract X-rays to a resolution limit of 3.01 A
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 9
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15 min, approximately 80% of the activity can be maintained in the range
658354
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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stable for at least 20 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
2-mercaptoethanol and phenol absorbing agents stabilize activity
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20% ethylene glycol and 0.2 M KCl stabilize activity
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KF prevents inactivation at room temperature
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Mg2+ stabilizes activity
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sensitive to freezing and thawing
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SPS-I loses more activity during purification than SPS-II
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-10°C, 2 months, 10% loss of activity
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-20°C, 1 month, no loss of activity, 1 year, 50-60% loss of activity
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-80°C, purified enzyme stable for at least 4 months
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0-4°C, stable in presence of 20% glycerol or 5 mM fructose-6-phosphate
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0°C, 4 weeks, 50% loss of activity
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4°C, stable for at least 1 week
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liquid nitrogen, stable for at least 11 months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
complete separation from sucrose synthetase
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enzyme becomes extremely unstable during the course of purification
-
gel filtration is carried out in presence of 20% ethylene glycol and 0.2 M KCl to stabilize activity; partial
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in presence of 2-mercaptoethanol and a phenol absorbing agent under N2; partial
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native isozymes partially by anion exchange chromatography
partial
partial, at least 3 additional proteins could not be removed
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partial, two forms of the enzyme identified, enzyme becomes extremely unstable during the course of purification
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partial, two forms of the enzyme identified, SPS-I loses more activity during purification than SPS-II
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transgenic SPS from tobacco and rice
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two forms of the enzyme identified
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
analysis of transient expression of SPS genes, functional significance of the various cis-acting regulatory elements present in banana SPS promoter in regulating SPS expression during ripening
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antisence transformants with reduced activity were produced
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expressed in tomato
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expressed in tomato and Escherichia coli, both reveal active enzyme, reduced amount of starch in leaves of transformed tomato
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expression in Escherichia coli
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expression in Escherichia coli and tobacco, active enzyme
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expression in Escherichia coli. The Synechocystis sps gene is introduced into tobacco, rice and tomato under the control of constitutive promoters. The Synechocystis SPS protein is expressed at high level. However SPS activities and carbon partitioning in leaves from transgenic and wild-type plants are not significantly different. The purified enzymes have full catalytic activity. It is proposed that some other protein in plant cells binds to the Synechocystis SPS resulting in inhibition of the enzyme
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expression in Populus alba x Populus grandidentata causing an altered phenotype compared to the wild-type trees with altered timing of bud flush and leaf senescence. Tree height and stem diameter are similar to the wild-type, but differences in the length of xylem fibres occur. Elevated concentrations of intracellular sucrose in both leaf and stem tissue of the transgenic trees are associated with a prolonged onset of senescence and an advancement in bud flush in the following spring, phenotypes, overview
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expression of a 26000 Da fragment in Escherichia coli
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gene SPS, cloning of SPS promoter region
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gene SPS, qualitative and quantitative realtime PCR analysis, overview
gene SPS1, real-time quantitative RT-PCR analysis of SPS1 expression in wild-type and transgenic plants
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gene SPS11, phylogenetic tree of the SPS gene family, quantitative expression analysis by real-time RT-PCR analysis; gene SPS1, phylogenetic tree of the SPS gene family, quantitative expression analysis by real-time RT-PCR analysis; gene SPS2, phylogenetic tree of the SPS gene family, quantitative expression analysis by real-time RT-PCR analysis; gene SPS6, phylogenetic tree of the SPS gene family, quantitative expression analysis by real-time RT-PCR analysis; gene SPS8, phylogenetic tree of the SPS gene family, quantitative expression analysis by real-time RT-PCR analysis
gene SPSII, semiquantitative PCR expression analysis
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genetic structure of SPSII genes, sequence comparisons and genetic mapping, phylogenetic tree, expression analysis of the SPSII family, overview
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Nicotiana tabacum cv. Xanthi plants are transformed with an arabidopsis SPS gene under the regulation of the ubiquitously expressed tandem repeat of the 35S cauliflower mosaic virus promoter. All transformed plants have significantly increased stem height, which is ascribed to internode elongation, and greater stem diameters, longer fibers and increased total dry biomass relative to the control plants. Difference in the chemical composition of either the storage or structural carbohydrates of the wild-type and SPS transgenic lines are only minor
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overexpression of maize sucrose-phosphate synthase gene in Nicotiana tabacum increases the sucrose synthesis and carbon assimilation, particularly in older leaves, accelerates the whole plant development and increases the abundance of flowers without substantial changes in the overall shoot biomass
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SPSA and SPSB, DNA and amino acid sequence determination and analysis
transgenic cotton over-producing spinach sucrose phosphate synthase shows enhanced leaf sucrose synthesis and improved fiber quality under controlled environmental conditions
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
ethylene treatment differentially stimulates SPS gene expression, a reverse GCCbox, ACCGCCG, ethylene responsive element is located within the SPS promoters of the three cultivars
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SPS activity is positively correlated with sucrose and negatively correlated with hexose sugars
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sucrose induces the enzyme, high sugar cultivars show increased transcript expression and enzyme activity of SPS compared to low sugar cultivars at all developmental stages. SPS activity is positively correlated with sucrose and negatively correlated with hexose sugars
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
sucrose phosphate synthase gene SPS is a suitable endogenous reference gene for genetically modified rice detection, method development and validation of the SPS gene as an endogenous reference gene and its optimized qualitative and quantitative PCR systems, overview
analysis
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sucrose-phosphate synthase is a biochemical marker of high sucrose accumulation in sugarcane
diagnostics
sucrose phosphate synthase gene SPS is a suitable endogenous reference gene for genetically modified rice detection, method development and validation of the SPS gene as an endogenous reference gene and its optimized qualitative and quantitative PCR systems, overview
additional information
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expression of the Arabidosis thaliana SPS gene in Populus alba x Populus grandidentata as model system for tree biology with substantial industrial relevance in the context of short rotation forestry and a target bioenergy crop
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