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
2.4.1.243
-
RECOMMENDED NAME
GeneOntology No.
6G-fructosyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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
-
-
-
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 (21)-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.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6(G)-fructosyltransferase/2,1-fructan:2,1-fructan 1-fructosyltransferase
P92916
bifunctional enzyme
6G-FFT
-
-
6G-FFT
Agave tequilana azul
-
-
-
6G-FFT
-
-
6G-FFT
P92916
-
6G-FFT
Q5FC15
-
6G-FFT
Q05613
-
6G-FFT
Lolium perenne Bravo
Q8S337
-
-
6G-FFT/1-FFT
Q8S337
-
AoFT1
Q5FC15
-
Atq6G-FFT-1
-
gene name
Atq6G-FFT-1
Agave tequilana azul
-
gene name
-
Atq6G-FFT-2
-
gene name
Atq6G-FFT-2
Agave tequilana azul
-
gene name
-
fructan-fructan 6G-fructosyltransferase
Q8S337
-
fructan: fructan 6G-fructosyltransferase
-
-
fructan: fructan 6G-fructosyltransferase
Agave tequilana azul
-
-
-
fructan:fructan 6G-fructosyltransferase
P92916
-
fructan:fructan 6G-fructosyltransferase
Q5FC15
-
fructan:fructan 6G-fructosyltransferase
Q05613
-
fructan:fructan 6G-fructosyltransferase
Q8S337
-
fructan:fructan 6G-fructosyltransferase
Lolium perenne Bravo
Q8S337
-
-
fructan:fructan 6G-fructosyltransferase/fructan:fructan 1-fructosyltransferase
Q8S337
-
CAS REGISTRY NUMBER
COMMENTARY
79633-28-6
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
genes Atq6G-FFT-1 and -2
-
-
Manually annotated by BRENDA team
Agave tequilana azul
genes Atq6G-FFT-1 and -2
-
-
Manually annotated by BRENDA team
bifunctional 6(G)-fructosyltransferase/2,1-fructan:2,1-fructan 1-fructosyltransferase
Swissprot
Manually annotated by BRENDA team
cv. Tenshin, enzyme activity remains stable during 16 weeks of storage at 15C, but decreases thereafter and increases during week 23 and 24 to almost original level
-
-
Manually annotated by BRENDA team
enzyme activity increases progrossively at 10C and at 20C during the first four months of storage and remains stable during the following two months. Acitivity at 10C is slightly higher than at 20C
-
-
Manually annotated by BRENDA team
expression in Cichorium intybus and Nicotiana tabacum
Swissprot
Manually annotated by BRENDA team
expression in Nicotiana tabacum
-
-
Manually annotated by BRENDA team
cultivar HLA-7
Swissprot
Manually annotated by BRENDA team
expression in Pichia pastoris, enzyme has slight fructan:fructan 1-fructosyltransferase activity
Swissprot
Manually annotated by BRENDA team
; perennial ryegrass
SwissProt
Manually annotated by BRENDA team
cultivar Aberystwyth S23
SwissProt
Manually annotated by BRENDA team
cultivars Aurora and Perma
UniProt
Manually annotated by BRENDA team
cv. Bravo
SwissProt
Manually annotated by BRENDA team
Lolium perenne Bravo
-
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
metabolism
Q8S337
the enzyme is required for fructan biosynthesis. In the second step, a 6G-FFT produces 6G-kestotriose, using 1-kestotriose as fructosyl donor and sucrose as fructosyl acceptor. In further steps, higher polymeric fructans are synthesized by a 1-FFT activity through beta(2-1)-linked chain elongation or by a 6-FT activity through beta(2-6)-linked chain elongation.The 6G-FFT from Lolium perenne also has 1-FFT activity
metabolism
-
the enzyme is required for fructan biosynthesis
metabolism
Agave tequilana azul
-
the enzyme is required for fructan biosynthesis
-
metabolism
Lolium perenne Bravo
-
the enzyme is required for fructan biosynthesis. In the second step, a 6G-FFT produces 6G-kestotriose, using 1-kestotriose as fructosyl donor and sucrose as fructosyl acceptor. In further steps, higher polymeric fructans are synthesized by a 1-FFT activity through beta(2-1)-linked chain elongation or by a 6-FT activity through beta(2-6)-linked chain elongation.The 6G-FFT from Lolium perenne also has 1-FFT activity
-
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 ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-kestose
?
show the reaction diagram
Agave tequilana, Agave tequilana azul
-
-
-
-
?
1-kestose + 1-kestose
?
show the reaction diagram
-
-
-
-
?
1-kestose + 1-kestose
?
show the reaction diagram
Q5FC15
-
-
-
?
1-kestose + 1-kestose
D-sucrose + 1G-kestotetraose + 6G-kestotetraose
show the reaction diagram
-, Q8S337
6G-FFT activity
-
-
?
1-kestose + 3 sucrose
3 neokestose + H2O
show the reaction diagram
P92916
-
-
-
?
1-kestose + 3 sucrose
3 neokestose + H2O
show the reaction diagram
Q8S337
-
-
-
?
1-kestose + 3 sucrose
3 neokestose + H2O
show the reaction diagram
-, Q8S337
-
-
-
?
1-kestose + 3 sucrose
3 neokestose + H2O
show the reaction diagram
Q05613
-
-
-
?
1-kestose + 3 sucrose
3 neokestose + H2O
show the reaction diagram
-, P92916
-
plus fructans of the inulin neoseries with a degree of polymerization up to six
-
?
1-kestose + 3 sucrose
3 neokestose + H2O
show the reaction diagram
Agave tequilana, Agave tequilana azul
-
-
plus a 4c tetrasaccharide moiety
-
?
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
D-sucrose + neokestose
show the reaction diagram
-, Q8S337
6G-FFT activity
-
-
?
1-kestose + sucrose
1-kestotetraose + 6-kestotetraose
show the reaction diagram
Lolium perenne, Lolium perenne Bravo
Q8S337
-
neither this fructan nor the subsequent mixed series is found in vivo in the tissues of Lolium perenne
-
?
1-kestose + sucrose
6G,6-kestotetraose
show the reaction diagram
Lolium perenne, Lolium perenne Bravo
Q8S337
-
-
-
?
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
Q5FC15
-
-
-
?
1F(1-beta-D-fructofuranosyl)3 sucrose + sucrose
1F,6G-di-beta-D-fructofuranosylsucrose + ?
show the reaction diagram
-
i.e. fructosyl nystose
-
-
?
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
-
r
neokestose + sucrose
1-kestose + ?
show the reaction diagram
-
-
reverse reaction, slow
-
r
neokestose + sucrose
1F,6G-di-beta-D-fructofuranosylsucrose + ?
show the reaction diagram
-
-
-
-
?
nystose + 1-kestose
?
show the reaction diagram
Q5FC15
-
-
-
?
sucrose
neokestose
show the reaction diagram
Agave tequilana, Agave tequilana azul
-
-
-
-
?
sucrose + sucrose
1F,6G-di-beta-D-fructofuranosylsucrose + ?
show the reaction diagram
-
-
-
-
?
sucrose + sucrose
1-kestotriose + ?
show the reaction diagram
-, Q8S337
-
-
-
?
4 1-kestose
sucrose + 1G-kestotetraose + 6G-kestotetraose + H2O
show the reaction diagram
-, Q8S337
-
-
-
?
additional information
?
-
-
plays an important role in the synthesis of inulin and inulinneo-series fructo-oligosaccharides
-
-
-
additional information
?
-
Q5FC15
main products are inulin neoseries fructan, such as 1F,6G-di-beta-D-fructofuranosylsucrose and neokestose
-
-
-
additional information
?
-
-, Q8S337
the enzyme possesses both fructan:fructan 6G-fructosyltransferase and fructan:fructan 1-fructosyltransferase activities
-
-
-
additional information
?
-
Q8S337
Lolium perenne plants produce a majority of levan neoseries built on the 6G-kestotriose backbone, incubation with both sucrose and 6G-kestotriose allow the synthesis of 6G,6-kestotetraose, both 1-kestotriose and 6G-kestotriose acts as fructosyl acceptors, producing 1- and 6-kestotetraose (bifurcose) and 6G,6-kestotetraose, respectively. 1-Kestotriose is not a fructosyl donor since incubation with 1-kestotriose alone does not lead to the synthesis of a 6-linked fructan. Bifurcose formation ceases and 6G,6-kestotetraose is formed instead, when recombinant fructan:fructan 6G-fructosyltransferase, 6G-FFT, is introduced in the enzyme assay with sucrose and 1-kestotriose as substrates
-
-
-
additional information
?
-
Agave tequilana, Agave tequilana azul
-
the Atq6GFFT-1 consensus sequence is DPCG, Atq6GFFT-1 and Atq6G-1FFT-2 utilize primarily 1-kestose as a substrate, a Asn/Trp couple is involved in substrate recognition
-
-
-
additional information
?
-
Lolium perenne Bravo
Q8S337
Lolium perenne plants produce a majority of levan neoseries built on the 6G-kestotriose backbone, incubation with both sucrose and 6G-kestotriose allow the synthesis of 6G,6-kestotetraose, both 1-kestotriose and 6G-kestotriose acts as fructosyl acceptors, producing 1- and 6-kestotetraose (bifurcose) and 6G,6-kestotetraose, respectively. 1-Kestotriose is not a fructosyl donor since incubation with 1-kestotriose alone does not lead to the synthesis of a 6-linked fructan. Bifurcose formation ceases and 6G,6-kestotetraose is formed instead, when recombinant fructan:fructan 6G-fructosyltransferase, 6G-FFT, is introduced in the enzyme assay with sucrose and 1-kestotriose as substrates
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
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
?
-
-
plays an important role in the synthesis of inulin and inulinneo-series fructo-oligosaccharides
-
-
-
additional information
?
-
Lolium perenne, Lolium perenne Bravo
Q8S337
Lolium perenne plants produce a majority of levan neoseries built on the 6G-kestotriose backbone, incubation with both sucrose and 6G-kestotriose allow the synthesis of 6G,6-kestotetraose
-
-
-
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Ag+
-
1 mM, no residual activity
Hg2+
-
1 mM, no residual activity
p-chloromercuribenzoate
-
0.1 mM, 2.5% residual activity
p-chloromercuribenzoate
-
-
Sodium dodecyl sulfate
-
1 mM, 48% residual activity
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Sodium deoxycholate
-
1 mM, 209% of initial activity
Triton X-100
-
1 mM, 160% of initial activity
Tween-80
-
1mM, 171% of initial activity
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
310
-
1F(1-beta-D-fructofuranosyl)2 sucrose
-
pH 5.7, 30C
88
-
1F(1-beta-D-fructofuranosyl)sucrose
-
pH 5.7, 30C
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
26.54
-
-
pH 5.68, 30C
136.8
-
-
pH 5.5, 30C
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.4
-
Q8S337
-
5.5
-
Q5FC15
-
5.7
-
-
-
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
6
Q8S337
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
-
more than 80% of maximum activity within this range
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
35
Q8S337
-
30
-
-
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
40
Q8S337
20C: about 80% of maximal activity, 40C: about 80% of maximal activity
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.11
-
-
Atq6G-FFT-2, sequence calculation
5.4
-
Q5FC15
calculated
5.57
-
-
Atq6G-FFT-1, sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Q8S337
the enzyme is predominantly expressed in the basal part of elongating leaves and leaf sheaths
Manually annotated by BRENDA team
Q8S337
elongating leaves, basal segment
Manually annotated by BRENDA team
P92916
induction by light
Manually annotated by BRENDA team
-
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
Lolium perenne Bravo
-
elongating leaves, basal segment
-
Manually annotated by BRENDA team
additional information
Q8S337
highly expressed in photosynthetically active tissues
Manually annotated by BRENDA team
additional information
-
age-related expression of Atq6G-FFT-1 in 5 and 7 year old plants, overview
Manually annotated by BRENDA team
additional information
Agave tequilana azul
-
age-related expression of Atq6G-FFT-1 in 5 and 7 year old plants, overview
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
66000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 52000, x * 25000, SDS-PAGE
?
-
x * 69000, SDS-PAGE
?
Q5FC15
x * 68311, calculated
?
-
x * 70700, about, Atq6G-FFT-1, sequence calculation, x * 69700, about, Atq6G-FFT-2, sequence calculation
?
Agave tequilana azul
-
x * 70700, about, Atq6G-FFT-1, sequence calculation, x * 69700, about, Atq6G-FFT-2, sequence calculation
-
additional information
-
N-terminal amino acid sequence of 52000 Da polypeptide and of 25000 Da polypeptide
additional information
-
Atq6G-FFT-1 and Atq6G-FFT-2 sequence comparisons
additional information
Agave tequilana azul
-
Atq6G-FFT-1 and Atq6G-FFT-2 sequence comparisons
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
Atq6G-FFT-1 and Atq6G-FFT-2 contains each seven glycosylation sites
glycoprotein
Agave tequilana azul
-
Atq6G-FFT-1 and Atq6G-FFT-2 contains each seven glycosylation sites
-
glycoprotein
Q5FC15
sequence contains six potential N-glycosilation sites
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.3
6.3
-
30C, stable for 30 min
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
40
-
stable for at least 15 min
37
-
Q5FC15
up to
45
-
-
20 min, pH 5.0-6.0, stable
50
-
-
inactivation
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
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
Q8S337
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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
Q8S337
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
genes Atq6G-FFT-1 and -2, DNA and amino acid sequence determination and analysis, expression in Pichia pastoris strain X-33
-
expressed in Lolium perenne genotypes N2 and F6
P92916
expressed in Pichia pastoris
Q8S337
expressed in Pichia pastoris strain X-33
Q8S337
FFT phylogenetic tree
Q8S337
heterologous expression in Pichia pastoris
Q8S337
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
N84A
-
preferred addition of fructose to a terminal fructose instead of terminal glucose, producing inulin-type fructans
N84A/S87N
-
prefers synthesis of beta(2-1) linkages
N84G
-
no enzymic activity
N84Q
-
preferred addition of fructose to a terminal fructose instead of terminal glucose, producing inulin-type fructans
N84S
-
preferred addition of fructose to a terminal fructose instead of terminal glucose, producing inulin-type fructans
N84Y
-
no enzymic activity
S87D
-
no change in substrate or product specificity
S87N
-
substrate and product specifitxy like wild-type
S87N
-
no change in substrate or product specificity
N340D/W343R
Q8S337
the mutant shows 6G-FFT and 1-FFT activities that hold the middle between those of the N340D and W343R single mutants
S415N
Q8S337
the mutant shows slightly lower 6G-FFT/1-FFT activities, but the ratio between the main reaction products is not significantly affected
additional information
-
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