Information on EC 2.4.1.100 - 2,1-fructan:2,1-fructan 1-fructosyltransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.4.1.100
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RECOMMENDED NAME
GeneOntology No.
2,1-fructan:2,1-fructan 1-fructosyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
[beta-D-fructosyl-(2->1)-]m + [beta-D-fructosyl-(2->1)-]n = [beta-D-fructosyl-(2->1)-]m-1 + [beta-D-fructosyl-(2->1)-]n+1
show the reaction diagram
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
fructan biosynthesis
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-
SYSTEMATIC NAME
IUBMB Comments
(2->1)-beta-D-fructan:(2->1)-beta-D-fructan 1-beta-D-fructosyltransferase
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CAS REGISTRY NUMBER
COMMENTARY hide
73379-55-2
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Agave inaequidens
-
-
-
Manually annotated by BRENDA team
var. azul
UniProt
Manually annotated by BRENDA team
Agave tequilana Weber var. azul
-
UniProt
Manually annotated by BRENDA team
burdock
-
-
Manually annotated by BRENDA team
strain HLA-7
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-
Manually annotated by BRENDA team
strain HLA-7
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
perennial ryegrass
SwissProt
Manually annotated by BRENDA team
Polymnia sonchifolia
-
-
-
Manually annotated by BRENDA team
Weber
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
physiological function
fructan accumulation occurs mainly in the first 2 weeks after anthesis, and a maximal fructan concentration of 2.5 mg fructan per kernel is reached at 16 days after anthesis. Fructan synthesis is catalyzed by 1-SST (sucrose:sucrose 1-fructosyltransferase) and 6-SFT (sucrose:fructan 6-fructosyltransferase), and to a lesser extent by 1-FFT (fructan:fructan 1-fructosyltransferase). Despite the presence of 6G-kestotriose in wheat kernel extracts, the measured 6G-FFT (fructan:fructan 6G-fructosyltransferase) activity levels are low
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
1,1,1-kestopentaose
inulin
show the reaction diagram
-
-
oligomers of up to DP 7
-
?
1,1-kestotetraose
inulin
show the reaction diagram
-
-
oligomers of up to DP 7
-
?
1,1-nystose + 1,1-nystose
sucrose + 1-kestose + oligofructan DP5 + oligofructan DP6
show the reaction diagram
-
-
-
?
1-kestose
1,1-nystose + ?
show the reaction diagram
-
-
-
-
?
1-kestose
1F,6G-di-beta-D-fructofuranosylsucrose + nystose
show the reaction diagram
1-kestose
?
show the reaction diagram
-
low affinity substrate
-
-
?
1-kestose + 1-kestose
1,1-nystose + 1,1,1-logose
show the reaction diagram
1-kestose + 1-kestose
D-sucrose + 1,1-nystose
show the reaction diagram
1-FFT activity
-
-
?
1-kestose + 6-kestose
1,6-nystose + 6,1-nystose
show the reaction diagram
-
-
-
?
1-kestotriose
1,1-kestotetraose + ?
show the reaction diagram
-
the enzyme transfers fructose moieties from and to 1-kestotriose or larger fructans
-
-
?
1-kestotriose
inulin
show the reaction diagram
-
-
oligomers of up to DP 6
-
?
1-kestotriose + 6G-kestotriose
1,1-kestotetraose + ?
show the reaction diagram
formation of 1,1-kestotetraose from 1-kestotriose as the single substrate by catalyzing this reaction by transferring fructose from a 1-kestotriose donor to a 6G-kestotriose acceptor
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-
?
1-kestotriose + sucrose
1,1-nystose
show the reaction diagram
the wild type enzyme clearly prefers 1-kestotriose over sucrose
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-
?
2 1-kestose
inulin + H2O
show the reaction diagram
-
-
up to DP 14
-
?
6-kestose + 1-kestose + sucrose
mixture of tetrasacharides of beta-2,6- and beta-2,1-linked fructans
show the reaction diagram
-
-
-
?
6G-kestose + 6G-kestose
?
show the reaction diagram
-
-
-
-
?
6G-kestose + sucrose
tetrasaccharide
show the reaction diagram
-
-
some synthesis of 1-kestose and resynthesis of 6G-kestose also occurs
?
D-Fructose
?
show the reaction diagram
-
low affinity substrate
-
-
?
fructose
?
show the reaction diagram
-
-
-
-
?
fructose + inulin
oligosaccharide
show the reaction diagram
-
-
reducing fructofuranosyl-only oligosaccharides
?
inulin
?
show the reaction diagram
-
high affinity substrate
-
-
-
nystose
1F(1-beta-D-fructofuranosyl)2-6G-beta-D-fructofuranosysucrose + 1F(1-beta-D-fructofuranosyl)3sucrose
show the reaction diagram
-
-
-
-
?
sucrose
?
show the reaction diagram
sucrose + 1,1-nystose
?
show the reaction diagram
-
-
-
-
?
sucrose + 1-kestose
1,1-nystose
show the reaction diagram
sucrose + 1-kestose
1-kestose + 6G-kestose + tetrasaccharide
show the reaction diagram
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-
transfer of fructosyl residue from 1-kestose to sucrose results in resynthesis of 1-kestose. Tetrasaccharides and 6G-kestose are also synthesized
?
sucrose + inulin
oligosaccharide
show the reaction diagram
sucrose + oligofructan
?
show the reaction diagram
-
-
-
-
?
[beta-D-fructosyl-(2->1)-]m + [beta-D-fructosyl-(2->1)-]n
?
show the reaction diagram
[beta-D-fructosyl-(2->1)-]m + [beta-D-fructosyl-(2->1)-]n
[beta-D-fructosyl-(2->1)-]m-1 + [beta-D-fructosyl-(2->1)-]n+1
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
[beta-D-fructosyl-(2->1)-]m + [beta-D-fructosyl-(2->1)-]n
?
show the reaction diagram
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
CuSO4
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1 mM, 22% inhibition
EDTA
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1 mM, 25% inhibition
p-chloromercuribenzoate
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0.1 mM, 97% inhibition
Sodium deoxycholate
Agave tequilana Weber var. azul
10 mM, 45% inhibition
Sodium dodecyl sulfate
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1 mM, 45% inhibition
Sucrose
additional information
-
cell wall/vacuolar inhibitor of fructosidase (0.001 mg) does not inhibit 1-FFT activity
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Sodium deoxycholate
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Triton X-100
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Tween-80
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
152
1,1-nystose
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18 - 119
1-kestose
233 - 377
1-kestotriose
976
D-sucrose
mutant enzyme N340D/W343R/S415N, in 50 mM sodium acetate buffer, pH 5.0, at 30°C
199
fructose
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440
nystose
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pH 5.5, 30°C
0.2 - 203
Sucrose
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
15
Sucrose
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.011
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in 50 mM Na-acetate, pH 5.2, and 0.02% Na-azide for 60 min at 30°C
11.63
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pH 5.5, 30°C
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 6.5
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5.6 - 8
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isoelectric focusing
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
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pH 5.0: about 70% of maximal activity, pH 8.0: about 65% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 40
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5°C: about 70% of maximal activity, 40°C: about 75% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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-
Manually annotated by BRENDA team
; developing from anthesis until maturity. 1-FFT is almost exclusively active during the first 9 days after anthesis and no 1,1-kestotetraose is produced during the last weeks of kernel development
Manually annotated by BRENDA team
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inner leaf base
Manually annotated by BRENDA team
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effect of nitrogen concentration
Manually annotated by BRENDA team
Polymnia sonchifolia
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-
Manually annotated by BRENDA team
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
17000
-
1 * 17000 + 1 * 52000, SDS-PAGE
19000
1 * 19000 + 1 * 52000, SDS-PAGE
49000
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x * 49000, SDS-PAGE
50000
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gel filtration
66000
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gel filtration
69000
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gel filtration
72800
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non-denaturing gel electrophoresis
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
-
N-terminal amino acid sequence
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.3 - 6.3
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30°C, stable for 30 min
660021
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 40
-
stable for 15 min
30
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1 h, stable
40
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1 h, up to 20% loss of activity below
50
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inactivation above
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
several attempts to purify 6G-FFT/1-FFT are unsuccessful, probably due to the instability of the enzyme and/or the rather low concentration in the yeast's expression medium
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
1°C, 8 weeks, an increase in 1-FFT activity is detected
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, several attempts (concanavalin A, Mono Q, Mono S) to further purify 6G-FFT/1-FFT are unsuccessful, probably due to the instability of the enzyme and/or the rather low concentration in the yeast’s expression medium
Fast Desalting column chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in leaves of Nicotiana benthamiana
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expressed in Pichia pastoris
expression in Pichia pastoris. The recombinant enzyme is a functional 1-FFT lacking invertase and sucrose:sucrose 1-fructosyl transferase activities, but shows a small intrinsic fructan exohydrolase activity
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
gene fft, sequence comparisons, recombinant expression of wild-type and mutant enzymes in Pichia pastoris
gene fft-1, sequence comparisons, recombinant expression in Pichia pastoris
heterologous expression in Pichia pastoris
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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elicitors such as salicylic acid and jasmonic acid methyl ester have a strong effect on fructooligosaccharide (FOS) accumulation. Exogenous application of 1 mM salicylic acid induces a 36fold accumulation of 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
expression of 1-FFT is strongly induced upon transfer to high-carbon/low-nitrogen medium, limiting nitrogen supply by lowering KNO3 and (NH4)2SO4 concentrations to 5 and 0.2 mM, respectively, under an elevated sucrose concentration (6%), results in a strong increase in transcript level for 1-FFT, 1-FFT expression is transiently and only moderately induced during the first 24 h of cold exposure
-
increasing the carbon availability by increasing the sucrose concentration from 3 to 6% (w/v) does not result in sustained 1-FFT induction, LY-294002 does not affect the induction of 1-FFT expression
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treatment with the calcium channel blocker LaCl3 prevents the induction of 1-FFT, the calmodulin antagonist W7 decreases the induction of 1-FFT, genistein and staurosporine reduce the expression of 1-FFT, endothal and cantharidin strongly repress the induction of 1-FFT in a very similar manner
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
N340D
the mutant shows only very low overall activity levels
N340D/S415N
the mutant shows similar properties to the W343R mutant, but the 1-nystose/neoketose ratio is decreased
N340D/W343R
the mutant shows 6G-FFT and 1-FFT activities that hold the middle between those of the N340D and W343R single mutants
N340D/W343R/S415N
the triple mutant gains the characteristic of using sucrose as a donor substrate, at the same time, this mutant can still use 1-kestose as a donor substrate, but in a much less efficient way than the wild type
S415N
the mutant shows slightly lower 6G-FFT/1-FFT activities, but the ratio between the main reaction products is not significantly affected
W343R
the mutant shows a wild type1-FFT activity but a largely reduced 6G-FFT activity
D281Y
the mutation restores the tight H-bond network and introduces typical S-type enzyme characteristics like producing similar amounts of 1-kestotriose compared to sucrose:sucrose 1-fructosyltransferases
H308T
site-directed mutagenesis
M19F
site-directed mutagenesis
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
nutrition
synthesis