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

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The expected taxonomic range for this enzyme is: Magnoliophyta

EC NUMBER
COMMENTARY hide
2.4.1.100
-
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
-
-
-
-
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
burdock
-
-
Manually annotated by BRENDA team
strain HLA-7
-
-
Manually annotated by BRENDA team
strain HLA-7
-
-
Manually annotated by BRENDA team
Colombia
-
-
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
Weber
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
Uniprot
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
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 + sucrose
1,1-nystose
show the reaction diagram
the wild type enzyme clearly prefers 1-kestotriose over sucrose
-
-
?
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
-
-
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
?
-
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
?
-
-
plays an important role in the synthesis of inulin and inulinneo-series fructo-oligosaccharides in onion bulb
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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 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 30C
199
fructose
-
-
440
nystose
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pH 5.5, 30C
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 30C
11.63
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pH 5.5, 30C
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
-
5C: about 70% of maximal activity, 40C: about 75% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
-
inner leaf base
Manually annotated by BRENDA team
-
effect of nitrogen concentration
Manually annotated by BRENDA team
Polymnia sonchifolia
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-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50000
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gel filtration
66000
-
gel filtration
69000
-
gel filtration
72800
-
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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30C, 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
-
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
1C, 8 weeks, an increase in 1-FFT activity is detected
-
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 yeasts expression medium
Fast Desalting column chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in leaves of Nicotiana benthamiana
-
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
heterologous expression in Pichia pastoris
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
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
-
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
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
nutrition
synthesis