Information on EC 3.2.1.157 - iota-carrageenase

Word Map on EC 3.2.1.157
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.2.1.157
-
RECOMMENDED NAME
GeneOntology No.
iota-carrageenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Endohydrolysis of (1->4)-beta-D-linkages between D-galactose 4-sulfate and 3,6-anhydro-D-galactose-2-sulfate in iota-carrageenans
show the reaction diagram
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
iota-carrageenan degradation
-
-
SYSTEMATIC NAME
IUBMB Comments
iota-carrageenan 4-beta-D-glycanohydrolase (configuration-inverting)
The main products of hydrolysis are iota-neocarratetraose sulfate and iota-neocarrahexaose sulfate. iota-Neocarraoctaose is the shortest substrate oligomer that can be cleaved. Unlike EC 3.2.1.81, beta-agarase and EC 3.2.1.83, kappa-carrageenase, this enzyme proceeds with inversion of the anomeric configuration. iota-Carrageenan differs from kappa-carrageenan by possessing a sulfo group on O-2 of the 3,6-anhydro-D-galactose residues, in addition to that present in the kappa-compound on O-4 of the D-galactose residues.
CAS REGISTRY NUMBER
COMMENTARY hide
74191-25-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Alteromonas fortis
gene cgiB_Ce; isolated from the red alga Grateloupia livida collected from Qingdao coastal waters of China Yellow Sea, gene cgiB_Ce
UniProt
Manually annotated by BRENDA team
gene cgiB_Ce; isolated from the red alga Grateloupia livida collected from Qingdao coastal waters of China Yellow Sea, gene cgiB_Ce
UniProt
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
iota-/nu-carrageenan + H2O
hydrolyzed iota/nu-carrageenan
show the reaction diagram
Alteromonas fortis
-
-
the smallest hybrid is an octasaccharide with a iota-iota-nu-iota structure, the second fraction is composed of two decasaccharides with iota-iota-iota-nu-iota and iota-[iota/nu]iota-iota structures, the third fraction is a mixture of dodecasaccharides which contains at least a iota-iota-iota-iota-nu-iota oligosaccharide
-
?
iota-carrageenan + H2O
?
show the reaction diagram
Alteromonas fortis
-
-
-
-
?
iota-carrageenan + H2O
hydrolyzed iota-carrageenan
show the reaction diagram
iota-carrageenan + H2O
iota-neocarratetraose sulfate + iota-neocarrahexaose sulfate
show the reaction diagram
iota-carrageenan + H2O
neo-iota-carratetraose
show the reaction diagram
iota-carrageenan + H2O
neo-iota-carratetraose + neo-iota-carrahexaose
show the reaction diagram
Alteromonas fortis
-
-
most abundant products
-
?
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
iota-carrageenan + H2O
hydrolyzed iota-carrageenan
show the reaction diagram
Alteromonas fortis
-
-
-
-
?
iota-carrageenan + H2O
neo-iota-carratetraose
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ba2+
118% activation at 1 mM
Cs+
in the presence of 0.1 M CsCl, 63% of maximal activity is observed
Fe2+
174% activation at 1 mM
Mg2+
149% activation at 1 mM
Mn2+
173% activation at 1 mM
NaCl
although its activity is enhanced by NaCl, the enzyme is active in the absence of NaCl
Ni2+
128% activation at 1 mM
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Al3+
56% inhibition at 1 mM
Cu2+
87% inhibition at 1 mM
EDTA
90% inhibition at 1 mM
Fe3+
11% inhibition at 1 mM
SDS
97% inhibition at 1 mM
Zn2+
49% inhibition at 1 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.2 - 8.5
iota-carrageenan
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4.1 - 726.6
iota-carrageenan
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.42 - 85.18
iota-carrageenan
7655
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.8
culture supernatant, at pH 7.5 and 50°C
80.6
after 10.3fold purification, at pH 7.5 and 50°C
1870
pH 6.5, 45°C, with iota-carrageenan; purified enzyme, pH 6.5, 45°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8
Alteromonas fortis
-
assay at, recombinant enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 11
activity range, overview
7 - 10
in a range of pH 7.0-10.0, the enzyme retains more than 80% of the original activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
Alteromonas fortis
-
-
50
in 50 mM Tris-HCl, pH 7.5, containing 0.1 M NaCl and 1 mM CaCl2
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 65
activity range, overview
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.79
sequence calculation
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
48071
x * 48071, sequence calculation; x * 48071, sequence calculation, x * 54000, recombinant His-tagged enzyme, SDS-PAGE
50000
-
gel filtration
55000
1 * 55000, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
concentration to 6 mg/ml using a dialyzing concentrator. The structure of iota-carrageenase bound to iota-carrageenan fragments is solved at 2.0 A resolution
Alteromonas fortis
hanging-drop vapour-diffusion method using polyethylene glycol (MW = 6000) as a precipitant. Crystals belong to space group P2(1), with unit-cell parameters a = 56.75 A, b = 91.04 A, c = 125.01 A, beta = 93.4°. The unit cell contains two molecules in the asymmetric unit related by a non-crystallographic twofold axis. Crystals diffracted to 2.0 A resolution on a synchrotron beamline
Alteromonas fortis
-
native and selenomethionyl-iota-carrageenase. Single crystals are obtained with polyethylene glycol, and the presence of Ca2+ appears to be crucial for crystallization. High quality crystals, typically 0.25 * 0.25 * 0.15 mM in dimension are grown with 0.1 M sodium cacodylate at pH 6.5, 15-17% polyethylene glycol and 200 mM calcium acetate. Crystallization of Se-Met-iota-carrageenase is performed under similar conditions except for the addition of 1 mM dithiothreitol and the replacement of sodium cacodylate by imidazole to avoid the reaction between cacodylate and dithiothreitol. Crystal structure at 1.6 A resolution
Alteromonas fortis
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9
purified recombinant His-tagged enzyme, 45°C, 24 h, stable
731522
6 - 9.6
stable at
731522
10
purified recombinant His-tagged enzyme, 45°C, 24 h, 60% activity remaining
731522
11
purified recombinant His-tagged enzyme, 45°C, 24 h, inactivation
731522
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
purified recombinant His-tagged enzyme, pH 6.5, 1 h, 80% activity remaining; purified recombinant His-tagged enzyme, pH 6.5, 1 h, inactivation; stable up to
45
the enzyme is stable up to 45°C in 50 mM Tris/HCl, pH 7.5, containing 0.1 M NaCl and 1 mM CaCl2 for 1 h. Incubation at 45°C inactivates the enzyme completely within 5 min at either condition of low NaCl concentration (less than 10 mM) or presence of 10 mM EDTA
50
purified recombinant His-tagged enzyme, pH 6.5, 1 h, 60% activity remaining
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme contains three calcium binding sites involved in stabilizing the enzyme structure
Alteromonas fortis
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, butyl-Toyopearl column chromatography, heparin affinity column chromatography, and Superdex 200 gel filtration
Ni-Sepharose column chromatography, Source 15S column chromatography, gel filtration
Alteromonas fortis
-
purified by affinity chromoatography
Alteromonas fortis
-
recombinant enzyme from Escherichia coli strain BL21(DE3) culture supernatant by affinity chromatography; recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Bacillus subtilis
expressed in Escherichia coli BL21(DE3) cells
Alteromonas fortis
-
expressed in Escherichia coli Origami (DE3) pLysS, BL21 (DE3), C41 (DE3), and C43 (DE3) cells
Alteromonas fortis
-
gene cgiB_Ce, DNA and amino acid sequence determination and analysis, phylogenetic analysis, recombinant expression in Escherichia coli strain BL21(DE3); gene cgiB_Ce, DNA and amino acid sequence determination and analysis, phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene encoding iota-carrageenase flanked by a C-terminal hexahistidine tag and a N-terminal PelB signal peptide for targeting the gene product into Escherichia coli periplasm. The recombinant plasmid, referred to as pETIAf, is used to transform Escherichia coli BL21(DE3) strain harbouring pLysS plasmid
Alteromonas fortis
-
overexpressed in Escherichia coli BL21(DE3)
Alteromonas fortis
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D247A
Alteromonas fortis
-
the enzymatic activity is completely abolished
E245Q
Alteromonas fortis
-
the enzymatic activity is completely abolished
E310Q
Alteromonas fortis
-
the Km value is reduced by a factor of about 4, the reduction in kcat by 50fold results in a substantial decrease in the overall enzyme efficiency (about 10fold)
H281A
Alteromonas fortis
-
the mutant shows 7fold decreased Km and 180fold decreased kcat value compared to the wild type enzyme with 4% relative efficiency
Q222E
Alteromonas fortis
-
the enzymatic activity is abolished
Q222K
Alteromonas fortis
-
the enzymatic activity is abolished
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
Alteromonas fortis
-
usage of the enzyme for the analysis of the distribution of the carrabiose moieties in hybrid carrageenan chains, hybrid iota-/alpha-carrageenan, by Pseudoalteromonas atlantica a 4S-iota carrageenan sulfatase that converts iota-carrabiose into alapha-carrabiose units