Information on EC 3.2.1.153 - fructan beta-(2,1)-fructosidase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.2.1.153
-
RECOMMENDED NAME
GeneOntology No.
fructan beta-(2,1)-fructosidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of terminal, non-reducing (2->1)-linked beta-D-fructofuranose residues in fructans
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
hydrolysis of O-glycosyl bond
-
-
SYSTEMATIC NAME
IUBMB Comments
beta-(2->1)-D-fructan fructohydrolase
Possesses one of the activities of EC 3.2.1.80, fructan beta-fructosidase. While the best substrates are the inulin-type fructans, such as 1-kestose [beta-D-fructofuranosyl-(2->1)-beta-D-fructofuranosyl alpha-D-glucopyranoside] and 1,1-nystose [beta-D-fructofuranosyl-(2->1)-beta-D-fructofuranosyl-(2->1)-beta-D-fructofuranosyl alpha-D-glucopyranoside], some (but not all) levan-type fructans can also be hydrolysed, but more slowly [see EC 3.2.1.154, fructan beta-(2,6)-fructosidase]. Sucrose, while being a very poor substrate, can substantially inhibit enzyme activity in some cases.
CAS REGISTRY NUMBER
COMMENTARY hide
1000593-08-7
-
37288-56-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
cv. Tenshin
-
-
Manually annotated by BRENDA team
cv. Morex
-
-
Manually annotated by BRENDA team
L. var. Bravo
SwissProt
Manually annotated by BRENDA team
enzyme contains three activities that hydrolyze beta-2,6-glycosidic linkages faster than beta-2,1-glycosidic linkages and two activities hydrolyze beta-2,1-glycosidic linkages faster than beta-2,6-glycosidic linkages
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
-
snow molds consume carbon sources contained in the inoculated wheat tissues but cannot effectively use wheat polysaccharide including fructans compared with mono- and disaccharides. Therefore, in snow mold inoculated wheat tissues, fructans are rapidly degraded, mainly by wheat enzymes, FEHs, to maintain necessary levels of mono- and disaccharides for metabolic demands under snow cover
physiological function
fructan 1-exohydrolase enzymes are involved in inulin degradation in the roots of chicory. Higher enzyme expression in cold temperatures can decrease the quality and the quantity of the inulin; fructan 1-exohydrolase enzymes are involved in inulin degradation in the roots of chicory. Higher enzyme expression in cold temperatures can decrease the quality and the quantity of the inulin; fructan 1-exohydrolase enzymes are involved in inulin degradation in the roots of chicory. Higher enzyme expression in cold temperatures can decrease the quality and the quantity of the inulin
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
1,1,1-kestopentaose + H2O
D-fructose + ?
show the reaction diagram
1,1-kestotetraose + H2O
?
show the reaction diagram
1,1-nystose + H2O
?
show the reaction diagram
1-kestose + H2O
?
show the reaction diagram
1-kestose + H2O
D-fructose
show the reaction diagram
from Vernonia herbacea and Vernonia discolor, but levans from Gomphrena macrocephala and Phleum pratense are poorly hydrolyzed, hydrolytic cleavage of terminal fructosyl residues off inulin, sucrose is not hydrolyzed
-
-
?
1-kestotriose + H2O
?
show the reaction diagram
1G-kestotetraose + H2O
?
show the reaction diagram
-
96% of the activity with 6G,1-kestotetraose
-
-
?
6-kestose + H2O
?
show the reaction diagram
6-kestotriose + H2O
?
show the reaction diagram
6G,1-kestotetraose + H2O
?
show the reaction diagram
-
-
-
-
?
6G,6-kestotetraose + H2O
?
show the reaction diagram
-
4% of the activity with 6G,1-kestotetraose
-
-
?
6G-kestotetraose + H2O
?
show the reaction diagram
-
-
-
-
?
6G-kestotriose + H2O
?
show the reaction diagram
-
5% of the activity with 6G,1-kestotetraose
-
-
?
inulin + H2O
?
show the reaction diagram
inulin + H2O
D-fructose
show the reaction diagram
5% inulin from Vernonia herbacea and Vernonia discolor, but levans from Gomphrena macrocephala and Phleum pratense are poorly hydrolyzed, hydrolytic cleavage of terminal fructosyl residues off inulin, sucrose is not hydrolyzed
-
-
?
inulin + H2O
D-fructose + ?
show the reaction diagram
-
-
-
?
inulin + H2O
fructose
show the reaction diagram
inulin + H2O
fructose + fructo-oligosaccharide + fructo-polysaccharide
show the reaction diagram
levan + H2O
?
show the reaction diagram
neokestose + H2O
?
show the reaction diagram
sucrose + H2O
D-fructose + D-glucose
show the reaction diagram
wheat graminan + H2O
?
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
inulin + H2O
D-fructose + ?
show the reaction diagram
Q93X60, Q9FNS9
-
-
-
?
wheat graminan + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
CaCl2
-
1 mM, partial
Na-EDTA
-
1 mM, partial
Sucrose
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
12
1,1,1-kestopentaose
-
-
8.3
1,1-kestotetraose
-
-
64
1,1-nystose
-
-
7 - 58
1-kestose
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
1-kestose
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.8 - 5.9
Sucrose
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0122
-
of 1-FEH I with 0.0083 micomol/min/mg background activity in Nicotiana benthamiana leaves
0.0205
-
of 1-FEH IIa with 0.0083 micomol/min/mg background activity in Nicotiana benthamiana leaves
12.2
-
isoenzyme 1-FEH w2
14.6
-
isoenzyme 1-FEH w1
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 6.5
0.05M citrate phosphate buffer, 30°C, at pH 5.5 maximal activity of more than 300 microgram fructose per mg protein and hour, at pH 4.5-5.0 about 90% activity, at pH 4.25 and 6.0 about 75% activity, at pH 4.0 and 6.5 about 60% activity
4.3 - 6.3
pH 4.3: about 60% of maximal activity, pH 6.3: about 50% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 35
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assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 50
at pH 5.5, maximal activity of slightly more than 400 microgram fructose per mg protein and h at 40°C, about 95% of maximal activity at 50°C, about 90% of maximal activity at 30°C, about 50% at 20°C, about 25% of maximal activity at 20°C
30 - 50
30C: about 70% of maximal activity, 50C: about 30% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.78
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calculation from cDNA, isoenzyme 1-FEH w2
4.79
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calculation from cDNA, isoenzyme 1-FEH w1
5.32
estimation by identification and characterization
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
-
activity is low in growing tubers but increases during dormancy and sprouting
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60000
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gel filtration
64000
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1 * 64000, SDS-PAGE
65700
calculated from sequence
70000
-
x * 70000, isoenzyme 1-FEH w1, SDS-PAGE; x * 70000, isoenzyme 1-FEH w2, SDS-PAGE
75000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 70000, isoenzyme 1-FEH w1, SDS-PAGE; x * 70000, isoenzyme 1-FEH w2, SDS-PAGE
monomer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
glycosylation on at least two of four potential N-glycosylation sites, isoenzyme 1-FEH w1; glycosylation on at least two of four potential N-glycosylation sites, isoenzyme 1-FEH w2
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method of recombinant enzyme heterologously expressed in Pichia pastoris. The structure of the isoenzyme 1-FEH IIa is determined at a resolution of 2.35 A. The structure consists of an N-terminal fivefold beta-propeller domain
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hanging-drop vapour-diffusion method at 4C. The crystals are tetragonal belonging to space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = 139.83, b = 139.83, c = 181.94 A
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
-
1 h, enzyme retains full activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
during storage at 15C, 1-FEH in onion bulbs is low but peaks abruptly after 12 and 16 weeks, respectively, after which it decreases to levels higher to that observed at the beginning of the storage
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
during storage at 15C, 1-FEH in onion bulbs is low but peaks abruptly after 12 and 16 weeks, respectively, after which it decreases to levels higher to that observed at the beginning of the storage
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
concentration by centrifugation after 4 d methanol induction
fructan 1-exohydrolase IIa and fructan 1-exohydrolase IIb
-
isoenzyme 1-FEH w1; isoenzyme 1-FEH w2
-
leaf material frozen, ground, soluble proteins extracted, precipitated, dialyzed, cell-wall associated proteins in pellets incubated in 1 M CaCl, supernatant precipitated, dialysed
-
partial, enzyme contains three activities that hydrolyze beta-2,6-glycosidic linkages faster than beta-2,1-glycosidic linkages and two activities hydrolyze beta-2,1-glycosidic linkages faster than beta-2,6-glycosidic linkages
-
precipitated, resuspended, desalted, centrifuged
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
48 h in Nicotiana benthamiana leaves infiltrated by Agrobacterium tumefaciens C58C1 with entry vector pDONR(TM)Zeo and destination vector pMDC32
-
enzyme expression analysis of isozymes under different water conditions, overview
-
expressed in Pichia pastoris; single copy gene Bp1-FEHa, DNA and amino acid sequence determination and analysis, phylogenetic tree, functional expression in Pichia pastoris
expression in Pichia pastoris
fructan 1-exohydrolase IIa and fructan 1-exohydrolase IIb
-
gene Wfh-sm3, recombinant expression in Pichia pastoris
-
heterologous expression in Pichia pastoris strain X-33 after culturing of plasmids in Escherichia coli, expression vector pPICZalphaA
heterologous expression in Pichia pastoris, pPICZalphaA vector
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isoenzyme 1-FEH w1; isoenzyme 1-FEH w2
-
quantitative real time PCR analysis of the FEH mRNAs (including FEHI and FEHII) in cold stored roots of three chicory cultivars, overview. Variable levels of FEHII and FEHI expression in different phenotypes of the chicory cultivars; quantitative real time PCR analysis of the FEH mRNAs (including FEHI and FEHII) in cold stored roots of three chicory cultivars, overview. Variable levels of FEHII and FEHI expression in different phenotypes of the chicory cultivars; quantitative real time PCR analysis of the FEH mRNAs (including FEHI and FEHII) in cold stored roots of three chicory cultivars, overview. Variable levels of FEHII and FEHI expression in different phenotypes of the chicory cultivars
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
expression of isozyme 1-FEH IIa is induced by cold stress (12 h at 4C); expression of isozyme 1-FEH I is induced by cold stress (12 h at 4C)
storage at low temperature (5C) enhances enzyme gene expression and activity
the profile of water-soluble carbohydrate accumulation and loss is negatively correlated with the mRNA concentration of 1-FEH, especially 1-FEH w3 (1-FEH-6B)
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
S101L
-
neighbouring amino acid exchange influences the orientation of Trp82, thus switches the function of sucrose as inhibitor towards hydrolysation substrate, but only when present in high concentrations because substrate binding is inefficient
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