Information on EC 3.2.1.65 - levanase

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

EC NUMBER
COMMENTARY hide
3.2.1.65
-
RECOMMENDED NAME
GeneOntology No.
levanase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Random hydrolysis of (2->6)-beta-D-fructofuranosidic linkages in (2->6)-beta-D-fructans (levans) containing more than 3 fructose units
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
-
-
-
-
hydrolysis of O-glycosyl bond
additional information
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Starch and sucrose metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
(2->6)-beta-D-fructan fructanohydrolase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9041-11-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
gene sacC
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Burkholderia caryophylli
-
-
-
Manually annotated by BRENDA team
var. Laevanivorans
-
-
Manually annotated by BRENDA team
no activity in Escherichia coli
-
-
-
Manually annotated by BRENDA team
Pseudomonas sp. No. 43
-
-
-
Manually annotated by BRENDA team
Pseudomonas strain K-52
-
-
-
Manually annotated by BRENDA team
Rhodotorula sp.
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
isolated from a Tunisian thermal source
-
-
Manually annotated by BRENDA team
yeasts
additional information
enzymatic activity from the sediment of the Gilau dam reservoir, Cluj county
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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 + H2O
D-fructose
show the reaction diagram
3 levan + 7 H2O
10 levanbiose
show the reaction diagram
Pseudomonas sp. No. 43
-
-
-
?
cellulose + H2O
?
show the reaction diagram
Pseudomonas strain K-52
-
-
-
-
?
D(+)-raffinose + H2O
beta-D-fructose + alpha-D-glucopyranosyl-(1-6)-alpha-D-glucopyranoside
show the reaction diagram
D(+)-raffinose + H2O
D-fructose
show the reaction diagram
-
-
-
?
D-(+)-xylose + H2O
?
show the reaction diagram
Pseudomonas strain K-52
-
-
-
-
?
dextran + H2O
?
show the reaction diagram
Pseudomonas strain K-52
-
-
-
-
?
fructose + H2O
?
show the reaction diagram
Pseudomonas strain K-52
-
-
-
-
?
galactose + H2O
?
show the reaction diagram
Pseudomonas strain K-52
-
-
-
-
?
glucose + H2O
?
show the reaction diagram
Pseudomonas strain K-52
-
-
-
-
?
inulin + H2O
?
show the reaction diagram
inulin + H2O
D-fructose
show the reaction diagram
inulin + H2O
D-fructose + ?
show the reaction diagram
lactose + H2O
?
show the reaction diagram
Pseudomonas strain K-52
-
-
-
-
?
levan + H2O
?
show the reaction diagram
levan + H2O
D-fructose
show the reaction diagram
levan + H2O
fructose
show the reaction diagram
levan + H2O
fructose + beta-1,2-oligofructans
show the reaction diagram
levan + H2O
levanbiose + D-fructose
show the reaction diagram
-
-
final products, thinlayer chromatography analysis
-
?
levan + H2O
levanheptaose
show the reaction diagram
-
-
-
?
levan + H2O
O-beta-D-fructofuranosyl-(2->6)-beta-D-fructofuranoside + ?
show the reaction diagram
levan + H2O
oligofructosides
show the reaction diagram
levan-type fructans (beta-2,6)
D-fructose
show the reaction diagram
-
-
-
-
?
levans + H2O
oligofructosides
show the reaction diagram
maltose + H2O
?
show the reaction diagram
Pseudomonas strain K-52
-
-
-
-
?
raffinose + H2O
?
show the reaction diagram
Pseudomonas strain K-52
-
-
-
-
?
soluble starch + H2O
?
show the reaction diagram
Pseudomonas strain K-52
-
-
-
-
?
sucrose + H2O
beta-D-fructofuranose + alpha-D-glucopyranose
show the reaction diagram
sucrose + H2O
D-fructose + D-glucose
show the reaction diagram
Timothy grass fructan + H2O
?
show the reaction diagram
xylan + H2O
?
show the reaction diagram
Pseudomonas strain K-52
-
-
-
-
?
levan + H2O
D-fructose
show the reaction diagram
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
?
show the reaction diagram
levan + H2O
?
show the reaction diagram
levan + H2O
fructose + beta-1,2-oligofructans
show the reaction diagram
levans + H2O
oligofructosides
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mn2+
-
slight activation at 75 mM
NaCl
-
slight activation at 75 mM
additional information
not significantly influenced by 1 mM Ca2+, Mg2+, or Fe3+, at 30C, pH 6.0, 1% levan, for 6 h
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
Pseudomonas sp. No. 43
-
-
Al3+
Pseudomonas sp. No. 43
-
-
Ammonium oxalate
ammonium persulfate
Rhodotorula sp.
-
-
ascorbic acid
CuSO4
1 mM, pH 6.0, 1% levan, for 6 h at 30C, 27% inhibition
diethyl dicarbonate
Pseudomonas strain K-52
-
-
glutathione
Rhodotorula sp.
-
-
Hydrazine sulfate
Rhodotorula sp.
-
-
iodoacetate
-
-
p-chloromercuribenzoate
Sodium citrate
Rhodotorula sp.
-
-
Sodium thioglycollate
Sucrose
-
weak inhibitory effect at high concentrations
Tris buffer
-
-
additional information
not inhibited by EDTA, dithiothreitol and beta-mercaptoethanol
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6.8
inulin
-
-
0.0012 - 0.97
levan
65
Sucrose
-
-
10
tetraose
-
-
-
additional information
additional information
Michaelis-Menten kinetics
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.09
-
U/ml, inulin
0.25
-
intracellular wild-type enzyme
0.3
-
BG4103-strain
0.33
-
extracellular mutant AD6 enzyme
0.4
-
QB167-strain
0.5
-
U/ml, inulin
0.52
-
extracellular mutant AD8 enzyme
0.69
-
intracellular mutant AD9 enzyme
0.71
-
extracellular mutant AD9 enzyme
0.73
-
intracellular mutant AD8 enzyme
0.75
-
intracellular mutant AD6 enzyme
1.8
purified recombinant enzyme, pH 6.0, 35C
14.24
-
extracellular wild-type enzyme
30.5
Pseudomonas sp. No. 43
-
U/mg
34.4
Rhodotorula sp.
-
-
37.9
crude recombinant enzyme, pH 6.5, 50C, substrate inulin
39.2
Pseudomonas strain K-52
-
-
58
-
levan, pH 5.0, at 30C
128
-
U/mg
226.7
-
purified native enzyme, pH 6.5, 40C
987
purified recombinant enzyme, pH 6.5, 50C, substrate inulin
4309
-
U/mg
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
not active below
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 7
inactive below pH 4.0, decreased activity above pH 7.0
5 - 7
-
levan, at 30C, activity decreases at pH 4.0 and at pH values above 7.0; sucrose, at 30C, activity decreases at pH 4.0 and at pH values above 7.0
5.5 - 6.5
-
in presence of levan extended to pH 4.6-7.5
5.5 - 7
higher activity, recombinant enzyme
5.5 - 6.5
Rhodotorula sp.
-
-
6 - 9
Pseudomonas strain K-52
-
4C
6 - 10
-
20C
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40 - 50
-
sucrose, pH 5.0
45
-
not active above 55C
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 60
-
activity range
30 - 40
Pseudomonas strain K-52
-
-
40 - 50
-
not active above 55C
45 - 55
-
-
50 - 70
activity range, profile overview
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
small extent transported into periplasmic space
Manually annotated by BRENDA team
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36000
Pseudomonas sp. No. 43
-
gel filtration
38000
Pseudomonas strain K-52
-
SDS-PAGE and gel filtration
39000
Rhodotorula sp.
-
SDS-PAGE
58400
calculation from sequence of cDNA
73000
-
SDS-PAGE
74000
-
in vitro transcription and translation experiments
75870
-
DNA sequence analysis
78000
-
SDS-PAGE
80000
-
gel filtration
86000
-
SDS-PAGE and DNA sequence analysis
89000
-
SDS-PAGE
100000
-
SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
no modification
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.6 - 7.5
-
-
136763
5 - 11
purified recombinant enzyme, 50C, 1 h, stable at
731185
5.5 - 6.5
Rhodotorula sp.
-
-
136768
6 - 8
Pseudomonas sp. No. 43
-
20C, stable up to 50C at pH 7.0; activity decreases rapidly below pH 6.0 and above pH 8.0
136770
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
Rhodotorula sp.
-
-
30 - 35
purified recombinant enzyme, 24 h, completely stable at
50
purified recombinant enzyme, pH 6.5, 1 h, stable up to
65
purified recombinant enzyme, pH 6.5, 1 h, inactivation
70
-
crude extracellular enzyme, pH 6.4, inactivation
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15C, several months
-
4C, phosphate buffer, pH 6.0
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation and Sephadex A-50 gel filtration
-
by immobilization-nickel affinity chromatography, centrifugation, column chromatography and dialysis
-
expression in Escherichia coli
-
native extracellular enzyme 397fold from culture supernatant by ammonium sulfate fractionation, dialysis, gel filtration, and anion exchange chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression in Escherichia coli
expression in B. subtilis
-
expression in Escherichia coli
expression in Lactobacillus plantarum
-
expression in Lactobacillus-Escherichia coli
-
expression in Pichia pastoris
gene lsdB, organized in one operon with gene lsdA, encoding levansucrase, organization of the lsdAB operon and relevant features of the intergenic sequence, expression analysis, overview
-
gene sacC, DNA and amino acid sequence determination and analysis, phylogenetic tree, recombinant expression in Escherichia coli strain BL21(DE3)
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
lsdB transcription is induced during growth with low fructose concentrations of 0.44 to 33 mM
lsdB transcription is repressed by D-glucose
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
medicine
nutrition
synthesis
analysis
additional information
-
the enzymatic activity can be used to characterize the aquatic ecosystems impurity degree, the evolution prognosis and water quality maintenance, the Gilau dam reservoir sediment has a decreased and quite unstable enzymatic potential
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