Information on EC 2.4.1.243 - 6G-fructosyltransferase

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The expected taxonomic range for this enzyme is: Liliopsida

EC NUMBER
COMMENTARY hide
2.4.1.243
-
RECOMMENDED NAME
GeneOntology No.
6G-fructosyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
[1-beta-D-fructofuranosyl-(2->1)-]m+1-alpha-D-glucopyranoside + [1-beta-D-fructofuranosyl-(2->1)-]n-alpha-D-glucopyranoside = [1-beta-D-fructofuranosyl-(2->1)-]m-alpha-D-glucopyranoside + [1-beta-D-fructofuranosyl-(2->1)-]n-beta-D-fructofuranosyl-(2->6)-alpha-D-glucopyranoside (m > 0; n >= 0)
show the reaction diagram
m > 0, n ? 0
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-
-
SYSTEMATIC NAME
IUBMB Comments
1F-oligo[beta-D-fructofuranosyl-(2->1)-]sucrose 6G-beta-D-fructotransferase
Inulins are polysaccharides consisting of linear or branched D-fructofuranosyl chains attached to the fructosyl residue of sucrose by a beta(2->1) linkage. This enzyme catalyses the transfer of the terminal (2->1)-linked -D-fructosyl group of an inulin chain onto O-6 position of the glucose residue of another inulin molecule [1]. For example, if 1-kestose [1F-(beta-D-fructofuranosyl)sucrose] is both the donor and recipient in the reaction shown above, i.e., if m = 1 and n = 1, then the products will be sucrose and 6G-di-beta-D-fructofuranosylsucrose. In this notation, the superscripts F and G are used to specify whether the fructose or glucose residue of the sucrose carries the substituent. Alternatively, this may be indicated by the presence and/or absence of primes (see {iupac/2carb/36#362::http://www.chem.qmul.ac.uk/iupac/2carb/36.html#362}). Sucrose cannot be a donor substrate in the reaction (i.e. m cannot be zero) and inulin cannot act as an acceptor. Side reactions catalysed are transfer of a beta-D-fructosyl group between compounds of the structure 1F-(1-beta-D-fructofuranosyl)m-6G-(1-beta-D-fructofuranosyl)n sucrose, where m >= 0 and n = 1 for the donor, and m >= 0 and n >= 0 for the acceptor.
CAS REGISTRY NUMBER
COMMENTARY hide
79633-28-6
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
genes Atq6G-FFT-1 and -2
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-
Manually annotated by BRENDA team
genes Atq6G-FFT-1 and -2
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-
Manually annotated by BRENDA team
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SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
additional information
-
the enzyme activity varies slightly during storage of fresh spears in the dark at 10C for 14 days, while the sugar content is rapidly lost, overview
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-kestose
?
show the reaction diagram
1-kestose + 1-kestose
?
show the reaction diagram
1-kestose + 1-kestose
D-sucrose + 1G-kestotetraose + 6G-kestotetraose
show the reaction diagram
6G-FFT activity
-
-
?
1-kestose + 3 sucrose
3 neokestose + H2O
show the reaction diagram
1-kestose + sucrose
1,1-kestotetraose + 1G-kestotetraose + 6G-kestotetraose
show the reaction diagram
-
-
prolonged incubation produces a complex fructan series with a higher degree of polymerization
-
?
1-kestose + sucrose
1-kestotetraose + 6-kestotetraose
show the reaction diagram
1-kestose + sucrose
6G,6-kestotetraose
show the reaction diagram
1-kestose + sucrose
D-sucrose + neokestose
show the reaction diagram
6G-FFT activity
-
-
?
1F(1-beta-D-fructofuranosyl)2 sucrose + sucrose
1F,6G-di-beta-D-fructofuranosylsucrose + ?
show the reaction diagram
-
i.e. nystose
-
-
?
1F(1-beta-D-fructofuranosyl)3 sucrose + sucrose
1F,6G-di-beta-D-fructofuranosylsucrose + ?
show the reaction diagram
1F(1-beta-D-fructofuranosyl)sucrose + sucrose
1F,6G-di-beta-D-fructofuranosylsucrose + ?
show the reaction diagram
-
i.e. 1-kestose
-
-
?
1F,6G-di-beta-D-fructofuranosylsucrose + sucrose
1-kestose + 1-kestose
show the reaction diagram
-
-
reverse reaction
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r
4 1-kestose
sucrose + 1G-kestotetraose + 6G-kestotetraose + H2O
show the reaction diagram
-
-
-
?
neokestose + sucrose
1-kestose + ?
show the reaction diagram
-
-
reverse reaction, slow
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r
neokestose + sucrose
1F,6G-di-beta-D-fructofuranosylsucrose + ?
show the reaction diagram
-
-
-
-
?
nystose + 1-kestose
?
show the reaction diagram
-
-
-
?
sucrose
neokestose
show the reaction diagram
sucrose + sucrose
1-kestotriose + ?
show the reaction diagram
-
-
-
?
sucrose + sucrose
1F,6G-di-beta-D-fructofuranosylsucrose + ?
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
1-kestose + 3 sucrose
3 neokestose + H2O
show the reaction diagram
Q8S337
-
-
-
?
4 1-kestose
sucrose + 1G-kestotetraose + 6G-kestotetraose + H2O
show the reaction diagram
Q8S337
-
-
-
?
additional information
?
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
p-chloromercuribenzoate
Sodium dodecyl sulfate
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1 mM, 48% residual activity
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Sodium deoxycholate
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1 mM, 209% of initial activity
Triton X-100
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1 mM, 160% of initial activity
Tween-80
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1mM, 171% of initial activity
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
310
1F(1-beta-D-fructofuranosyl)2 sucrose
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pH 5.7, 30C
88
1F(1-beta-D-fructofuranosyl)sucrose
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pH 5.7, 30C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26.54
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pH 5.68, 30C
136.8
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pH 5.5, 30C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 6
pH 5.0: about 80% of maximal activity, pH 6.0: about 90% of maximal activity, pH 4.3: about 20% of maximal activity, pH 6.5: about 20% of maximal activity
5.4 - 6.3
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more than 80% of maximum activity within this range
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 40
20C: about 80% of maximal activity, 40C: about 80% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.11
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Atq6G-FFT-2, sequence calculation
5.57
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Atq6G-FFT-1, sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
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Manually annotated by BRENDA team
induction by light
Manually annotated by BRENDA team
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the enzyme activity varies slightly during storage of fresh spears in the dark at 10C for 14 days, while the sugar content is rapidly lost, overview
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
66000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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
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stable for at least 15 min
45
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20 min, pH 5.0-6.0, stable
50
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inactivation
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 yeasts expression medium
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
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Lolium perenne genotypes N2 and F6
expressed in Pichia pastoris
expressed in Pichia pastoris strain X-33
FFT phylogenetic tree
genes Atq6G-FFT-1 and -2, DNA and amino acid sequence determination and analysis, expression in Pichia pastoris strain X-33
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heterologous expression in Pichia pastoris
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
N84A
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preferred addition of fructose to a terminal fructose instead of terminal glucose, producing inulin-type fructans
N84A/S87N
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prefers synthesis of beta(2-1) linkages
N84G
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no enzymic activity
N84Q
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preferred addition of fructose to a terminal fructose instead of terminal glucose, producing inulin-type fructans
N84S
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preferred addition of fructose to a terminal fructose instead of terminal glucose, producing inulin-type fructans
N84Y
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no enzymic activity
S87D
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no change in substrate or product specificity
N340D/W343R
the mutant shows 6G-FFT and 1-FFT activities that hold the middle between those of the N340D and W343R single mutants
S415N
the mutant shows slightly lower 6G-FFT/1-FFT activities, but the ratio between the main reaction products is not significantly affected
additional information
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exchange of N-terminal 36 amino acids with those of sucrose:sucrose 1-fructosyltransferase and vacuolar invertase. The latter two enzyme no longer used sucrose as a substrate, but were able to use 1-kestose