Information on EC 2.4.1.99 - sucrose:sucrose fructosyltransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.4.1.99
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RECOMMENDED NAME
GeneOntology No.
sucrose:sucrose fructosyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2 sucrose = D-glucose + beta-D-fructofuranosyl-(2->1)-beta-D-fructofuranosyl alpha-D-glucopyranoside
show the reaction diagram
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-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
fructosyl group transfer
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-
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hexosyl group transfer
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-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
fructan biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
sucrose:sucrose 1'-beta-D-fructosyltransferase
For definition of the prime in the systematic name, see 2-Carb-36.2.
CAS REGISTRY NUMBER
COMMENTARY hide
73379-56-3
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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-
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Manually annotated by BRENDA team
Agave inaequidens
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-
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Manually annotated by BRENDA team
strain Weber var. azul
UniProt
Manually annotated by BRENDA team
Agave tequilana Weber var. azul
-
UniProt
Manually annotated by BRENDA team
Agave vera cruz
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-
-
Manually annotated by BRENDA team
Agropyron sp.
Agropyron cristatum Gaertn. * Agropyron desertorum Schult
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-
Manually annotated by BRENDA team
strain HLA-7
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-
Manually annotated by BRENDA team
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-
-
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
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-
-
Manually annotated by BRENDA team
Crinum longifolium
-
-
-
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
timothy grass
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-
Manually annotated by BRENDA team
Polymnia sonchifolia
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-
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Manually annotated by BRENDA team
strain LEB-V10
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-
Manually annotated by BRENDA team
strain LEB-V10
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-
Manually annotated by BRENDA team
cultivar Prima
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
fungal FOS-synthesizing enzymes are sucrose-sucrose 1-fructosyltransferases (1-SST) with a conserved catalytic cavity representing a chimeric sequence from plant invertases and transferases
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
1,1-nystose + 1,1-nystose
1,1,1-fructosylnystose + 1-kestose
show the reaction diagram
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self-transfructosylation at a low rate
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?
1-kestose
nystose + D-fructose
show the reaction diagram
1-kestose + 1-kestose
1,1-nystose + sucrose
show the reaction diagram
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self-transfructosylation at a low rate
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?
1-kestose + 1-kestose
D-fructose + sucrose + ?
show the reaction diagram
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and formation of some nystose
-
?
6-kestose
bifurcose
show the reaction diagram
-
-
-
?
neo-kestose
1G-kestotetraose + 6G-kestotetraose
show the reaction diagram
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-
-
?
sucrose + 1-kestose
D-glucose + ?
show the reaction diagram
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-
-
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sucrose + 6G(1-beta-fructofuranosyl)n-sucrose
D-glucose + 1F-beta-fructofuranosyl-6G(1-beta-fructofuranosyl)2-sucrose
show the reaction diagram
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-
-
?
sucrose + H2O
D-fructose + D-glucose
show the reaction diagram
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-
-
-
?
sucrose + neokestose
D-glucose + 1F,6G-di-beta-fructofuranosylsucrose
show the reaction diagram
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-
-
?
sucrose + sucrose
1-kestose
show the reaction diagram
sucrose + sucrose
1-kestose + D-glucose
show the reaction diagram
sucrose + sucrose
1-kestose + nystose + fructosylnystose
show the reaction diagram
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18.7% (w/w) 1-kestose, 35.6% (w/w) nystose and 6.4% (w/w) fructosylnystose. The rest of carbohydrates in the mixture are fructose (minoritary), glucose and remaining sucrose
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?
sucrose + sucrose
D-glucose + 1-kestose
show the reaction diagram
sucrose + sucrose
D-glucose + 1F-beta-D-fructosylsucrose
show the reaction diagram
sucrose + sucrose
D-glucose + 6-kestose
show the reaction diagram
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FT1 has high affinity for 6-kestose and low affinity for 1-kestose
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-
?
sucrose + sucrose
D-glucose + ?
show the reaction diagram
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-
-
?
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
sucrose + sucrose
D-glucose + 1-kestose
show the reaction diagram
sucrose + sucrose
D-glucose + 1F-beta-D-fructosylsucrose
show the reaction diagram
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
activates
Co2+
-
activates
Cu2+
Agave vera cruz
-
activates
CuSO4
-
121% activity at 10 mM
Li+
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activates
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
CaCl2
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69% residual activity at 10 mM
choline chloride
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Citric acid
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65% residual activity at 10 mM
Co2+
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0.25 mM CoCl2, 12.7% inhibition
CsCl2
Agropyron sp.
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iodoacetamide
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1 mM, 10% inhibition
KCl
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40% residual activity at 10 mM
MnCl2
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99% residual activity at 10 mM
NaCl
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74% residual activity at 10 mM
NEM
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1 mM, 10% inhibition
Sodium deoxycholate
Agave tequilana Weber var. azul
10 mM, 32% inhibition
UDP
Agave vera cruz
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ZnSO4
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68% residual activity at 10 mM
additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cysteine
Agave vera cruz
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activates
pyridoxal-HCl
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20 mM, stimulation to 147% of the control
pyridoxine
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20 mM, stimulation to 120% of the control
SDS
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1.5fold activation at 10 mM
Sodium deoxycholate
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1.4fold activation at 1 mM
Triton X-100
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1.4fold activation at 5% (w/v)
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.083 - 551
Sucrose
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4.3 - 16200
Sucrose
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.054
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in 50 mM Na-acetate, pH 5.2, and 0.02% Na-azide for 60 min at 30C
0.5
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crude extract
75.4
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after 150.8fold purification
231
pH 5.5, 30C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7
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substrate sucrose
5.7
Agave vera cruz
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.7 - 6.5
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about 50% of maximal activity at pH 2.7 and at pH 6.5
4 - 8
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pH 4.0: about 50% of maximal activity, pH 8.0: about 79% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 25
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synthesis of 1-kestose
37
Agave vera cruz
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65 - 70
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0 - 30
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the native enzyme shows minor sucrose:sucrose 1-fructosyltransferase side activities at 0C but higher sucrose:sucrose 1-fructosyltransferase side activities at 30C
5 - 37
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50% of maximal activity at 5C and at 37C, synthesis of 1-kestose
40 - 75
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40C: about 60% ofmaximal activity, 75C: about 80% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.02
calculated
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
; developing from anthesis until maturity. When sucrose is the sole substrate, 1-kestotriose formation is highest during the first 9 days after anthesis, as well as 1-SST activity for 1-kestotriose formation from sucrose and 6G-kestotriose, after which the latter activity suddenly decreases. Little 1-kestotriose formation is seen in wheat kernels harvested between 9 and 28 days after anthesis whereas no such activity is observed later on during kernel development. 1+6G-kestotetraose formation follows a similar pattern to 1-kestotriose synthesis throughout kernel development although the formed 1+6G-kestotetraose levels are always lower
Manually annotated by BRENDA team
the relative expression of the fructan biosynthetic genes 1-SST and 6-SFT, encoding sucrose:sucrose 1-fructosyltransferase and sucrose:fructan 6-fructosyltransferase, respectively, and 6G-FFT/1-FFT isoform 6G-FFT 2 correlates positively with the accumulation of fructans in both blades and pseudostems. Isoform 6G-FFT 2 is highly expressed in cultivar E, that accumulatss the highest levels of fructans in this study
Manually annotated by BRENDA team
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after 1 month of growth the activity is twice as high as in N-rich plants
Manually annotated by BRENDA team
the relative expression of the fructan biosynthetic genes 1-SST and 6-SFT, encoding sucrose:sucrose 1-fructosyltransferase and sucrose:fructan 6-fructosyltransferase, respectively, and 6G-FFT/1-FFT isoform 6G-FFT 2 correlates positively with the accumulation of fructans in both blades and pseudostems. Isoform 6G-FFT 2 is highly expressed in cultivar E, that accumulatss the highest levels of fructans in this study
Manually annotated by BRENDA team
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at the flowering stage
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
Polymnia sonchifolia
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higher activitiy values are found at the beginning of the tuberization, 3-months-old plants, and at the flowering phase, 7 months-old plants
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26000
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1 * 26000 + 1 * 59000, SDS-PAGE
49000
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SDS-PAGE
50000
x * 61800, calculated, x * 50000 and x * 52000, SDS-PAGE
52000
x * 61800, calculated, x * 50000 and x * 52000, SDS-PAGE
59000
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1 * 26000 + 1 * 59000, SDS-PAGE
61500
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x * 61500, calculation from nucleotide sequence
61800
x * 61800, calculated, x * 50000 and x * 52000, SDS-PAGE
62000
Agave vera cruz
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gel filtration
67000
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gel filtration
68000
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gel filtration
75000
x * 75000, SDS-PAGE
77000
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2 * 77000, SDS-PAGE
170000
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gel filtration
180000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
computational model based on fructan 1-exohydrolase IIa, PDB entry 1st8
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5
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24 h, about 65% loss of activity
674979
4 - 8
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30C, stable
489628
4
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24 h, about 50% loss of activity
674979
5.5
optimal stability at
689660
5.5 - 7.5
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24 h, stable, pH stability is assayed in the presence of high concentration of a carbohydrate (600 g/l alpha-D-methylglucopyranoside)
674979
8
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24 h, about 20% loss of activity
674979
9
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24 h, about 30% loss of activity
674979
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 37
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10 min, stable
25
optimal stability at
30
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pH 4.0-8.0, 30 min, stable
35
rapid inactivation above
50 - 60
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10 min, inactivation
55
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24 h, stable up to
65
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24 h, about 50% loss of activity
66
-
very stable at temperatures below 66C, the half-life of the enzyme decreases very quickly at temperatures above 66C
100
-
the enzyme is inactivated at 100C after 10 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
operational stability in continous culture is 30 days
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operational stability in semibatch culture 20-60 days, operational stability in continous culture is 100 days
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several attempts to purify 1-SST are unsuccessful, probably due to the instability of the enzyme and/or the rather low concentration in the yeast's expression medium
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, several attempts (concanavalin A, Mono Q, Mono S) to further purify 1-SST are unsuccessful, probably due to the instability of the enzyme and/or the rather low concentration in the yeast's expression medium
partial
Q sepharose column chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli DH5alpha cells
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expressed in leaves of Nicotiana benthamiana
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expressed in Lolium perenne
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expressed in Oryza sativa
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expressed in Pichia pastoris
expression in Pichia pastoris
expression in Saccharomyces cerevisiae
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expression in tobacco protoplasts and in Pichia pastoris
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gene 1-FFT, semi-quantitative reverse transcriptase-PCR enzyme expression analysis
Agave inaequidens
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gene 1-SST, semi-quantitative reverse transcriptase-PCR enzyme expression analysis
subcloned into a pUC18 derivative and expressed in tobacco protoplasts under the control of the cauliflower mosaic virus 35S RNA promotor
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
0.2 mM jasmonic acid methyl ester induces a 85fold accumulation of fructooligosaccharide (FOS) in the stems of Agave inaequidens. Significant FOS accumulation in this species also occurs in response to treatments with 1 mM salicylic acid, 8% sucrose, and 10% polyethylene glycol
Agave inaequidens
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exogenous application of 1 mM salicylic acid induces a 36fold accumulation of fructooligosaccharide (FOS) of various degrees of polymerization in stems. 50 mM abscisic acid, 8% sucrose, and 1.0 mg/L kinetin also lead to a significant accumulation of low and high degrees of polymerization FOS in this species
FT1 transcription is readily induced under high sucrose or cold conditions associated with increased sucrose:sucrose 6-fructosyltransferase activity
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mRNA levels of the 1-SST gene first increases, but then decreases before increasing again during a longer period of cold treatment
plants at the vegetative stage present high 1-sucrose:sucrose fructosyltransferase in the distal regionof the rhizophores
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SST-1 is drastically diminished when cold (5C) and/or excision are imposed
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the enzyme is down-regulated during the 2 to 4 days after imbibition germination
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C203N
the mutant has an almost doubled relative hydrolytic activity in comparison to wild type enzyme
L322F
the mutant shows wild type activity
M173L
the mutant shows wild type activity
W362Y
the mutant has an almost doubled relative hydrolytic activity in comparison to wild type enzyme
Y248V
the mutant shows no activity
Y317W
the mutant shows wild type activity
Y408W
the mutant shows low activity
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
nutrition
synthesis
-
high transferase/hydrolase ratio confers it a great potential for the industrial production of prebiotic fructooligosaccharides