Information on EC 3.2.1.80 - fructan beta-fructosidase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
3.2.1.80
-
RECOMMENDED NAME
GeneOntology No.
fructan beta-fructosidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of terminal, non-reducing (2->1)- and (2->6)-linked beta-D-fructofuranose residues in fructans
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
hydrolysis of O-glycosyl bond
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
fructan degradation
-
-
Fructose and mannose metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
beta-D-fructan fructohydrolase
Hydrolyses inulin and levan, and also sucrose.
CAS REGISTRY NUMBER
COMMENTARY hide
37288-56-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
T14V
-
-
Manually annotated by BRENDA team
va Tenshin
-
-
Manually annotated by BRENDA team
(Fr.) Keissler
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
JNSP5-06
-
-
Manually annotated by BRENDA team
AF10
SwissProt
Manually annotated by BRENDA team
strain SL-09
-
-
Manually annotated by BRENDA team
oat
-
-
Manually annotated by BRENDA team
strain 11
-
-
Manually annotated by BRENDA team
strain 11
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
G7a1
-
-
Manually annotated by BRENDA team
Jerusalem artichoke
-
-
Manually annotated by BRENDA team
barley
-
-
Manually annotated by BRENDA team
B235
-
-
Manually annotated by BRENDA team
gene Lp6-FEHa
UniProt
Manually annotated by BRENDA team
cultivar Hokushu
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
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
1,1,1-kestopentaose + H2O
?
show the reaction diagram
-
18% activity compared to levan
-
-
?
1,1,1-kestopentaose + H2O
beta-D-fructose + ?
show the reaction diagram
-
-
-
-
?
1,1,1-kestose + H2O
beta-D-fructose + sucrose
show the reaction diagram
1,1-kestose + H2O
beta-D-fructose + sucrose
show the reaction diagram
1,1-kestotetraose + H2O
?
show the reaction diagram
1,1-nystose + H2O
beta-D-fructose + sucrose
show the reaction diagram
1-kestose + H2O
?
show the reaction diagram
1-kestose + H2O
beta D-fructofuranose + sucrose
show the reaction diagram
1-kestose + H2O
beta-D-fructose + sucrose
show the reaction diagram
1-kestotriose + H2O
?
show the reaction diagram
1-nystose + H2O
?
show the reaction diagram
-
-
-
-
?
6,6-kestotetraose + H2O
?
show the reaction diagram
6-kestose + H2O
?
show the reaction diagram
-
2% activity compared to 1-kestose
-
-
?
6-kestose + H2O
beta-D-fructose + sucrose
show the reaction diagram
6-kestotriose + H2O
?
show the reaction diagram
6G,6-kestotetraose + H2O
D-fructose + neokestose
show the reaction diagram
-
100%, chicorium inulin 11.9%, Aerobacter levan 8.5%, sucrose 0% rate of hydrolysis
-
?
6G-kestotriose + H2O
?
show the reaction diagram
Actilight + H2O
?
show the reaction diagram
-
highest activity, Actilight is a commercial mixture composed of 4.8% (w/v) of free sugars (sucrose, glucose and fructose), 35.1% w/v of 1-kestose, 53% w/v of 1-nystose and 7.1% w/v of 1-fructofuranosyl-nystose
-
-
?
Agave tequilana fructan + H2O
?
show the reaction diagram
-
-
-
-
?
burdock root sugar extract + H2O
fructose + sucrose + ?
show the reaction diagram
-
-
-
-
?
fructan + H2O
beta-D-fructose + ?
show the reaction diagram
fructans + H2O
D-fructose
show the reaction diagram
fructosylnystose + H2O
?
show the reaction diagram
-
97% activity compared to 1-kestose
-
-
?
garlic extract + H2O
?
show the reaction diagram
inulin + 5 H2O
5 beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
inulin + H2O
2 1-Kestose
show the reaction diagram
-
-
-
-
?
inulin + H2O
?
show the reaction diagram
inulin + H2O
beta-D-fructose
show the reaction diagram
inulin + H2O
beta-D-fructose + ?
show the reaction diagram
inulin + H2O
beta-D-fructose + sucrose
show the reaction diagram
inulin + H2O
beta-D-Fruf-(2-1)-beta-D-Fruf
show the reaction diagram
-
-
-
-
?
inulin + H2O
D-fructose
show the reaction diagram
inulin + H2O
D-fructose + D-glucose
show the reaction diagram
inulin + H2O
D-fructose + D-glucose + fructooligosaccharides
show the reaction diagram
-
a typical solution product consists of a mixture of fructose (155 g/l), glucose (155 g/l), sucrose (132 g/l) and fructooligosaccharides (50 g/l)
-
-
?
inulin + H2O
D-fructose + fructooligosaccharides
show the reaction diagram
inulin + H2O
D-glucose + D-fructose
show the reaction diagram
inulin + H2O
oligofructosides
show the reaction diagram
inulin + H2O
sucrose
show the reaction diagram
-
-
-
-
?
inulin-type fructans (beta-2,1) + H2O
D-fructose
show the reaction diagram
-
poorly substrate
-
-
?
inulobiose + H2O
?
show the reaction diagram
-
12% activity compared to 1-kestose
-
-
?
inulobiose + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
inulin is an intermediate product
-
-
?
inuloheptaose + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
inulohexaose, inulopentaose, inulotetraose, inulotriose, inulobiose and inulin are intermediate products
-
-
?
inulohexaose + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
inulopentaose, inulotetraose, inulotriose, inulobiose and inulin are intermediate products
-
-
?
inulopentaose + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
inulotetraose, inulotriose, inulobiose and inulin are intermediate products
-
-
?
inulotetraose + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
inulotriose, inulobiose and inulin are intermediate products
-
-
?
inulotriose + H2O
?
show the reaction diagram
-
41% activity compared to 1-kestose
-
-
?
inulotriose + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
inulobiose and inulin are intermediate products
-
-
?
levan + H2O
?
show the reaction diagram
levan + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
-
-
-
?
levan + H2O
beta-D-fructose
show the reaction diagram
levan + H2O
beta-D-fructose + ?
show the reaction diagram
levan + H2O
D-fructose
show the reaction diagram
levan + H2O
D-fructose + ?
show the reaction diagram
levan-type fructan + H2O
beta-D-fructose + ?
show the reaction diagram
-
-
-
-
?
levan-type fructans (beta-2,6) + H2O
D-fructose
show the reaction diagram
-
best substrate
-
-
?
levanbiose + H2O
beta-D-fructose + ?
show the reaction diagram
-
-
-
-
?
levanohexaose + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
levanopentaose, levanotetraose, levanotriose, levanobiose and levan are intermediate products
-
-
?
levanopentaose + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
levanotetraose, levanotriose, levanobiose and levan are intermediate products
-
-
?
levanotetraose + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
levanotriose, levanobiose and levan are intermediate products
-
-
?
levanotriose + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
levanobiose and levan are intermediate products
-
-
?
neokestin + H2O
D-fructose
show the reaction diagram
-
-
-
?
neokestose + H2O
?
show the reaction diagram
-
7% activity compared to 1-kestose
-
-
?
neokestose + H2O
beta-D-fructose + ?
show the reaction diagram
neokestose + H2O
beta-D-fructose + sucrose
show the reaction diagram
-
4% activity of isoform w1 and 3% of w2 in comparison to substrate 1-kestose
-
-
?
neokestose + H2O
D-fructose + sucrose
show the reaction diagram
-
-
-
?
nystose + H2O
?
show the reaction diagram
-
66% activity compared to 1-kestose
-
-
?
phlein + H2O
beta-D-fructose + ?
show the reaction diagram
-
-
-
-
?
raffinose + H2O
beta-D-fructofuranose + O6-alpha-D-galactopyranosyl-alpha-D-glucopyranoside
show the reaction diagram
raffinose + H2O
beta-D-fructose + alpha-D-galactopyranosyl-1,6-alpha-D-glucopyranose
show the reaction diagram
-
-
-
-
?
raffinose + H2O
beta-D-fructose + alpha-D-galactopyranosyl-1,6-alpha-D-glucopyranoside
show the reaction diagram
-
-
-
-
?
raffinose + H2O
D-fructose + alpha-D-galactopyranosyl-1,6-alpha-D-glucopyranoside
show the reaction diagram
stachyose + H2O
beta-D-fructose + alpha-D-galactopyranosyl-1,6-alpha-D-galactopyranosyl-1,6-alpha-D-galactopyranose
show the reaction diagram
-
-
-
-
?
sucrose + H2O
alpha-D-glucose + beta-D-fructose
show the reaction diagram
-
3% enzyme activity in comparison to levan
-
-
?
sucrose + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
sucrose + H2O
beta-D-fructose + alpha-D-glucose
show the reaction diagram
sucrose + H2O
beta-D-fructose + D-glucose
show the reaction diagram
sucrose + H2O
D-fructose + D-glucose
show the reaction diagram
wheat graminan + H2O
beta-D-fructose + ?
show the reaction diagram
-
high degree of polymerization
-
-
?
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
fructans + H2O
D-fructose
show the reaction diagram
inulin + 5 H2O
5 beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
inulin + H2O
oligofructosides
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ba2+
-
30 min at 30ºC, 10 mM, 140% of the relative activity without metal ions; 30 min at 30ºC, 1 mM, 118% of the relative activity without metal ions; 30 min at 30ºC, 5 mM, 132% of the relative activity without metal ions
Cu2+
-
activating at concentrations of 1.0 mM
K+
-
activating at concentrations of 1.0 mM
Mg2+
-
30 min at 30ºC, 1 mM, 110% of the relative activity without metal ions
Na+
-
activating at concentrations of 1.0 mM
Zn2+
-
activating
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
-
strong inhibitor, after preincubation of the purified enzyme for 1 h at 0°C
2,5 dideoxy-2,5-imino-D-mannitol
-
-
Al3+
at 55ºC, pH 4.5, 5 mM, 89% inhibition
Ba2+
-
more than 50% of residual enzymatic activity at 20 mM
CaCl2
-
partially
CuSO4
-
1 mM, 47% inhibition
D-fructose
-
-
D-glucose
Fe2(SO4)3
-
1 mM, 38% inhibition
iodoacetate
-
30 min at 30ºC, 10 mM, 87% inhibition; 30 min at 30ºC, 1 mM, 38% inhibition; 30 min at 30ºC, 5 mM, 57% inhibition
iodoacetic acid
-
strong inhibitor, after preincubation of the purified enzyme for 1 h at 0°C
N-bromosuccinimide
1.0 mM, pH 5.0, at 30ºC, 100% inhibition
p-chloromercuribenzoate
phenylmethanesulfonyl fluoride
-
strong inhibitor, after preincubation of the purified enzyme for 1 h at 0°C
SDS
-
final concentration of 10.0 mM
Sodium arsenate
-
30 min at 30ºC, 10 mM, 50% inhibition; 30 min at 30ºC, 1 mM, 20% inhibition; 30 min at 30ºC, 5 mM, 38% inhibition
Sodium arsenite
-
30 min at 30ºC, 10 mM, 58% inhibition; 30 min at 30ºC, 1 mM, 28% inhibition; 30 min at 30ºC, 5 mM, 41% inhibition
Sucrose
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
DDT
at 55ºC, pH 4.5, 5 mM, 121% of the relative activity without metal ions
EDTA
-
slight stimulatuion after 30 min of incubation
KCN
-
slight stimulatuion after 30 min of incubation
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.6 - 7
1-kestose
27.8
Actilight
-
purified enzyme, at pH 5.0 and 55°C
-
12.9
Agave tequilana fructan
-
purified enzyme, at pH 5.0 and 50°C
-
0.05 - 66
inulin
3.72
Inulobiose
at 37ºC, pH 4.5
1.1
inuloheptaose
at 37ºC, pH 4.5
1.02
inulohexaose
at 37ºC, pH 4.5
1.08
inulopentaose
at 37ºC, pH 4.5
0.94
inulotetraose
at 37ºC, pH 4.5
1.63
inulotriose
at 37ºC, pH 4.5
1.8
levan
-
purified enzyme, at pH 5.0 and 50°C
1.01
levanohexaose
at 37ºC, pH 4.5
10.2
levanopentaose
at 37ºC, pH 4.5
2.74
levanotetraose
at 37ºC, pH 4.5
0.21
levanotriose
at 37ºC, pH 4.5
2.7
raffinose
-
pH 5.5, 50°C
25.3 - 111
Sucrose
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
21
1-kestose
Arabidopsis thaliana
Q67XZ3
in 50 mM sodium acetate buffer, pH 5.0, at 30°C
1.09
Actilight
Kluyveromyces marxianus
-
purified enzyme, at pH 5.0 and 55°C
-
0.9
Agave tequilana fructan
Kluyveromyces marxianus
-
purified enzyme, at pH 5.0 and 50°C
-
0.1 - 1110
inulin
0.33
levan
Kluyveromyces marxianus
-
purified enzyme, at pH 5.0 and 50°C
1.27 - 5000
Sucrose
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.09
Actilight
Kluyveromyces marxianus
-
purified enzyme, at pH 5.0 and 55°C
41694
0.9
Agave tequilana fructan
Kluyveromyces marxianus
-
purified enzyme, at pH 5.0 and 50°C
41693
0.1
inulin
Kluyveromyces marxianus
-
purified enzyme, at pH 5.0 and 55°C
677
0.18
levan
Kluyveromyces marxianus
-
purified enzyme, at pH 5.0 and 50°C
2411
1.27
Sucrose
Kluyveromyces marxianus
-
purified enzyme, at pH 5.0 and 55°C
55
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.5
Sucrose
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.2
-
reaction mixture consists of 0.1 ml purified inulinase, 0.9 ml acetate buffer, 20 g/l inulin; incubation at 50°C for 10 min; reaction inactivated at 100°C for 10 min
1.765
-
enzyme from supernatant, at pH 6.5 and 30°C
4.02
-
crude extract, at pH 5.0 and 50°C
12.2
-
isoform 1-FEH w1
13.65
-
native protein, pH 5.0
14.6
-
isoform 1-FEH w1
26.6
-
protein expressed in Pichia pastoris, pH 5.0
40.4
-
substrate: inulin, recombinant InuE
49.5
-
after 12.3fold purification, at pH 5.0 and 50°C
73.4
-
after 41.6fold purification, at pH 6.5 and 30°C
93.8
sucrose, at 40ºC, pH 5.0
150
-
substrate: sucrose, recombinant InuE
156
at 55ºC, pH 4.5
171
-
substrate: sucrose, InuE:D' hybrid
280
-
substrate: inulin,InuE:D' hybrid
743
inulin, at 40ºC, pH 5.0
12300
-
pH 5.0, 50°C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 6.5
-
for Agave tequilana fructan as substrate, the enzymatic activity is constant from pH 4.0 to 6.5
4.5 - 5.5
-
neokestin
4.5 - 5
-
using levan as substrate
4.5 - 5.5
-
both isoforms w1 and w2
5 - 5.5
-
inulin
6.5 - 7
-
at pH 8.0, 58% of maximal activity
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5 - 7
-
little fructanase activity at or above pH 7.0
4 - 7
-
pH 4.0: about 40% of maximal activity, pH 7.0: about 45% of maximal activity
4 - 6
-
85% of maximal activity at both pH 4.0 and 6.0
4 - 6.5
-
above and below rapid decrease of enzyme activity
4 - 7
-
pH 4.0: about 20% of maximal activity, pH 7.0: about 45% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 40
-
both isoforms w1 and w2
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 35
-
65% of maximal activity at 15°C, 70% of maximal activity at 35°C
20 - 55
-
rapid inactivation at 60°C
40 - 65
-
40°C: about 55% of maximal activity, 65°C: about 65% of maximal activity
40 - 70
40 - 65
-
40°C: about 65% of maximal activity, 65°C: about 55% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.6 - 4.7
-
isoelectric focusing
5
-
calculated from amino acid sequence
additional information
-
-
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
1-FEH activity is high only in induced sprouting; spatial and temporal distribution, mainly in the distal region of the rhizophore at vegetative phase and in the proximal region at sprouting phase
Manually annotated by BRENDA team
additional information
-
no activity in leaves
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
33000
-
x * 33000, SDS-PAGE
43000
-
x * 43000, SDS-PAGE
50000
-
x * 50000, deglycosylated recombinant enzyme, SDS-PAGE
59400
-
1 * 66000 + 1 * 62700 + 1 * 59400, SDS-PAGE
62700
-
1 * 66000 + 1 * 62700 + 1 * 59400, SDS-PAGE
63000
-
SDS-PAGE
63400
-
x * 63400, calculated from amino acid sequence
65666
-
x * 65666, calculated from amino acid sequence
66000
-
1 * 66000 + 1 * 62700 + 1 * 59400, SDS-PAGE
73924
-
x * 73924, InuE:D hybrid, calculated from sequence
74520
calculation from sequence of amino acid
78000
-
x * 78000, SDS-PAGE
81000
SDS-PAGE
83000 - 85000
-
gel filtration, SDS-PAGE
110000
-
x * 110000, InuE:D hybrid, SDS-PAGE
115000
-
gel filtration
125000
126000
-
gel filtration
127000
-
x * 127000, SDS-PAGE
140000
-
x * 140000, SDS-PAGE
150000 - 200000
gel filtration
176500
-
native PAGE
200000
-
gel filtration
250000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterotrimer
-
1 * 66000 + 1 * 62700 + 1 * 59400, SDS-PAGE
homodimer
-
2 * 125000, SDS-PAGE
monomer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
no modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
enzyme-fructose-complex, 1.87 A resolution, single isomorphous replacement, 20% fructose solution; free enzyme, 1.5 A resolution, single isomorphous replacement, orthorhombic crystals, 0.5 M sodium iodide and flash-cooling to -173°C in stream of cold nitrogen
-
2.35 A resolution, hanging drop vapour diffusion, structure determination by single anomalous dispersion method
hanging-drop vapour diffusion method, crystallization of complexes between 1-FEH IIa with the substrate 1-kestose and the inhibitors sucrose, fructose and 2,5-dideoxy-2,5-imino-D-mannitol; mutant proteins are crystallized by hanging drop vapour diffusion method with 2.1 M sodium potassium phosphate and 0.1 M potassium phosphate, pH 7.0
-
X-ray diffraction study
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5
-
55% of maximal activity
657381
4 - 5
-
at 55ºC
655714
4 - 9.5
-
-
654218
4 - 6.5
-
rapid decrease of residual activity above and below, after treatment at pH 6.0 and 4°C for 2 h still 96.0% residual activity
690488
4.5 - 10
-
30°C, 24 h, InuE:D', stable
680950
4.5
-
almost complete loss of activity at
208818
5 - 6
-
50°C, 3 h, stable
681357
5 - 7
at 30ºC
655019
5.4
-
stable for 60 min without decrease in activity, stable for 12 h with 96% residual activity, stable for 72 h with 63.8% residual activity
690509
5.5 - 9
-
-
714715
5.5 - 7.5
-
95-100% of maximal activity retained
657429
7
-
37°C, 30 min
208818
7.5
-
above pH 7.5 the isoforms are inactive
657048
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
more than 70% of maximal activity at pH 5.8 after 24 h
4 - 50
20% of the maximal activity is still detected at 4°C, 1-FEHa activity decreases strongly above 40°C and falls to 20% of ist maximal activity at 50°C
4
-
the activity of the enzyme remains relatively high even at 4°C, with up to 38% of its activity at 30°C
15
-
more than 70% of maximal activity at pH 5.8 after 24 h
25
-
more than 70% of maximal activity at pH 5.8 after 24 h
45
-
30 min, inulinase activity of InuE:D' is stable below
50 - 70
pH 5.0, enzyme remains stable up to 50ºC, but it loses activity at 70ºC
60 - 80
-
the recombinant immonbilized enzyme retains 88%, 77%, and 57% of its initial activity after preincubation for 10 days at temperatures of 60°C, 70°C, and 80°C, respectively. The soluble enzyme is inactivated at a higher rate, overview
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
freezing and thawing accelerates inactivation
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
10°C, 10 weeks, triggered by decrease in sucrose content
-
20°C, 10 weeks, triggered by decrease in sucrose content
-
4°C, 0.02 mg/ml, 20 mM phosphate buffer, pH 7.0, stable at least for several days, 90% loss of activity after 2 weeks
-
50°C, pH 5.4, stable for 60 min without decrease in activity, after 12 h only 96% residual activity, after 72 h only 63.8% residual activity
-
65°C, 2 h, 86% residual activity after treatment at 65°C, pH below 4.0 and above 6.5 results in rapid decrease in enzyme activity
-
at either -15ºC or at 4ºC, 200 mM acetate buffer, pH 4.6, 2 weeks, 50% loss of activity
-
higher stability at -20°C than at 4°C
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
4.2-fold purification, 21% yield
-
7.2-fold purification, analyzed by SDS-PAGE, 2.44-fold increase of specific inulinase activity with 22.4% yield after DEAE sepharose fast flow anion exchange chromatography
-
ammonium sulfate precipitation, ConA affinity-, ion exchange-chromatography, and gel filtration
-
by ammonium sulfate precipitation, DEAE-Sephacel column, Sephacryl S-200 column, concanavalin A-linked amino-activated silica column and Sepharose 6B column, 25fold
-
by ammonium sulfate precipitation, Sephadex column and DEAE-cellulose column, 62fold
-
by ultrafiltration, DEAE-cellulofine column, Q-Sepharose column and Superdex column, 329fold
centrifugation, concentration and desaltation with 20 mM citrate-phosphate buffer pH 5.0 on Amicon Ultra-15 concentrator
-
DEAE-Sepharose CL-6B column chromatography, Toyopearl HW55S column chromatography, and Toyopearl HW55S column chromatography
-
purification of recombinant inuE and inuE:D' fusion protein. The internal region D' encoding the 157-amino-acid sequence in the Penicillium exoinulinase gene inuD cDNA is inserted into the site between the nucleotides 897 and 898 of the Aspergillus niger exoinulinase gene inuE cDNA. The resultant inuE:D' fusion is expressed in Pichia pastoris and purified
-
recombinant enzyme, by ammonium sulfate precipitation and anion exchange column, 4.8fold
recombinant extracellular enzyme from Pichia pastoris strain X33 from the culture supernatant
recombinant extracellular His-tagged enzyme from Pichia pastoris strain PpBfrA(4X) culture supernatant by nickel affinity chromatography
-
Sephadex G-25
-
Sephadex G50 gel filtration
two independent isoforms 1-FEH IIa and 1-FEH IIb from leaves and roots
-
two isoforms 1-FEH w1 and w2
-
ultrafiltration and Mono Q Sepharose column chromatography
-
ultrafiltration, 1.3fold
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a modified Bacillus subtilis levanase gene (sacC) in which the native bacterial signal sequence is replaced with a yeast alpha-factor domain is expressed in Saccharomyces cerevisiae strain AH22
enzyme expression analysis of isozymes under different water conditions, overview
-
expressed in Escherichia coli Top10 cells
-
expressed in Pichia pastoris
expressed in Pichia pastoris strain X33
expressed in Pichia pastoris; expression in Pichia pastoris
-
expression in Escherichia coli
expression in Pichia pastoris
expression in potato tubers
-
gene Lp6-FEHa, DNA and amino acid sequence determination and analysis, phylogenetic analysis, real-time quantitative PCR expression analysis, cloning in Escherichia coli, recombinant expression in Pichia pastoris strain X33, secretion of the protein to the culture medium
heterologous expression in Pichia pastoris
heterologous expression in Pichia pastoris X-33
-
in Escherichia coli
recombinant expression of the C-terminally His-tagged extracellular enzyme in Pichia pastoris strain PpBfrA(4X), secretion to the culture medium, fed-batch fermentation
-
the enzyme gene promoter is highly expressed in etiolated leaves and cold-stored roots
the internal region D' encoding the 157-amino-acid sequence in the Penicillium exoinulinase gene inuD cDNA is inserted into the site between the nucleotides 897 and 898 of the Aspergillus niger exoinulinase gene inuE cDNA. The resultant inuE:D' fusion is expressed in Pichia pastoris
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
in the stubble of seedlings and in young haplocorms from adult timothy plants, enzyme transcripts are significantly increased within 3 h of defoliation
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E201Q
-
inactive
P294N
-
mutant with an introduced N-glycosylation site near the inulin-binding cleft, shows highly decreased activity against higher DP inulin. The introduction of a glycosyl chain most probably blocks the cleft and prevents inulin binding and degradation; shows highly decreased activity against inulin
W82L
-
inactive; no inhibition by sucrose
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
diagnostics
-
expression of 1-FEH w3 may be a good indicator to identify potential grain yield in wheat exposed to water deficits during grain filling, when linked to osmotic potential and green leaf retention
food industry
nutrition
synthesis
additional information
Show AA Sequence (250 entries)
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