Information on EC 3.2.1.83 - kappa-carrageenase

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

EC NUMBER
COMMENTARY hide
3.2.1.83
-
RECOMMENDED NAME
GeneOntology No.
kappa-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 in kappa-carrageenans
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
kappa-carrageenan degradation
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-
SYSTEMATIC NAME
IUBMB Comments
kappa-carrageenan 4-beta-D-glycanohydrolase (configuration-retaining)
The main products of hydrolysis are neocarrabiose-sulfate and neocarratetraose-sulfate [5]. Unlike EC 3.2.1.157 (iota-carrageenase), but similar to EC 3.2.1.81 (beta-agarase), this enzyme proceeds with retention of the anomeric configuration.
CAS REGISTRY NUMBER
COMMENTARY hide
37288-59-8
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain Dsij, isolated from Delesseria sanguinea
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-
Manually annotated by BRENDA team
Calcalispora hiemalis
-
-
-
Manually annotated by BRENDA team
isolated from the sediment of carrageenan production base in Hainan (Hainan, China)
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-
Manually annotated by BRENDA team
isolated from the sediment of carrageenan production base in Hainan (Hainan, China)
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-
Manually annotated by BRENDA team
Culicidospora gravida
-
-
-
Manually annotated by BRENDA team
Cytophaga sp. 1k-C783
1k-C783
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Dactylella aquatica
-
-
-
Manually annotated by BRENDA team
Dendrospora erecta
-
-
-
Manually annotated by BRENDA team
Leske
-
-
Manually annotated by BRENDA team
Gyoerffyella speciosa
-
-
-
Manually annotated by BRENDA team
Heliscella stellata
-
-
-
Manually annotated by BRENDA team
Lateriramulosa uni-inflata
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
no activity in Tricladium angulatum
-
-
-
Manually annotated by BRENDA team
isolated from seaside soil under the stacks of red algae in Hainan province of China
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-
Manually annotated by BRENDA team
isolated from seaside soil under the stacks of red algae in Hainan province of China
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-
Manually annotated by BRENDA team
Pleuropedium tricladioides
-
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
Pseudomonas carrageenovora
Pyramidospora densa
-
-
-
Manually annotated by BRENDA team
Taeniospora gracilis
-
-
-
Manually annotated by BRENDA team
Tamlana sp.
-
-
-
Manually annotated by BRENDA team
Tamlana sp. HC4
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-
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Manually annotated by BRENDA team
Tricellula aquatica
-
-
-
Manually annotated by BRENDA team
Zobellia sp.
i.e. strain ZM-2, gene cgkZ
UniProt
Manually annotated by BRENDA team
i.e. strain ZM-2, gene cgkZ
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
agar + H2O
?
show the reaction diagram
iota-carrageenan + H2O
?
show the reaction diagram
kappa-carrageen + H2O
?
show the reaction diagram
kappa-carrageen + H2O
carrageenan oligosaccharide
show the reaction diagram
kappa-carrageen + H2O
kappa-neocarrahexaose-sulfate + kappa-neocarrabiose-sulfate + kappa-neocarrahexaose-sulfate
show the reaction diagram
kappa-carrageenan + H2O
?
show the reaction diagram
kappa-carrageenan + H2O
kappa-neocarrabiose sulfate + D-galactose
show the reaction diagram
kappa-carrageenan + H2O
kappa-neocarrabiose sulfate + kappa-neocarratetraose sulfate + kappa-neocarrahexaose sulfate + kappa-neocarraoctaose sulfate + kappa-neocarradecaose sulfate + D-galactose
show the reaction diagram
kappa-carrageenan + H2O
tetrasaccharides
show the reaction diagram
lambda-carrageenan + H2O
?
show the reaction diagram
p-nitrophenyl-beta-D-galactopyranoside + H2O
?
show the reaction diagram
-
-
-
-
-
kappa-carrageenan + H2O
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
kappa-carrageen + H2O
?
show the reaction diagram
kappa-carrageenan + H2O
?
show the reaction diagram
-
degradation
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ba2+
-
116.5% activity at 10 mM
Fe2+
-
105.9% activity at 10 mM
Mg2+
-
119% activity at 10 mM
NaCl
-
optimal activity at 150 mM
additional information
Zobellia sp.
-
poor effects by Li+, Mg2+, NH4+, Ca2+ and K+ at 5 mM
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
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63.3% residual activity at 5 mM
AgNO3
-
1 mM, complete inhibition
AlCl3
-
1 mM, 39.7% inhibition
BaCl2
-
1 mM, 26.8% inhibition
CaCl2
-
1 mM, 50.7% inhibition
CuSO4
-
1 mM, 31.3% inhibition
FeCl3
-
1 mM, 36.5% inhibition
Hg2+
-
3% residual activity at 10 mM
iodoacetic acid
-
79.7% residual activity at 5 mM
Li+
-
83.1% residual activity at 10 mM
Mg2+
Tamlana sp.
-
65.93% residual activity at 1 mM
NaI
-
strong inhibition at 200 mM, inhibition is caused by iodide binding
Ni2+
-
9.7% residual activity at 10 mM
Pb2+
Zobellia sp.
-
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phenylmethylsulfonyl fluoride
-
58.6% residual activity at 5 mM
Tris
-
strong inhibition at 200 mM
ZnCl2
-
1 mM, 50% inhibition
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
DTT
Zobellia sp.
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-
Triton X-100
Zobellia sp.
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Tween 80
Zobellia sp.
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.023
kappa-carrageen
Zobellia sp.
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pH 6.0, 39C, recombinant enzyme
-
0.244
kappa-carrageenan
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-
-
additional information
additional information
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1
ZnCl2
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0774
Tamlana sp.
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cell-free culture supernatant, in 25 mM Tris-HCl (pH 8.0) at 30C
6
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measurement of production of reducing sugar
6.192
Tamlana sp.
-
after 80.38fold purification, in 25 mM Tris-HCl (pH 8.0) at 30C
8.16
purified recombinant enzyme, pH 6.0, 50C
19.8
-
crude enzyme, at pH 8.0, 55C
61
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cell-free culture supernatant, in 25 mM Tris-HCl (pH 8.0) at 32C
700.5
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purified enzyme, pH 7.0, 40C
1170
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purified enzyme, pH 7.0, 35C
4040
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after 202.6fold purification, at pH 8.0, 55C
5778
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after 95fold purification, in 25 mM Tris-HCl (pH 8.0) at 32C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.6
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and a second optimum at pH 7.7
7.2
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in Tris buffer
7.5
Pseudomonas carrageenovora
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-
7.7
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and a second optimum at pH 5.6
8.8
recombinant enzyme, at 30C
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 9.5
Zobellia sp.
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activity range
4.8 - 9
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pH 4.8: about 70% of maximal activity, pH 9.0: about 60% of maximal activity
5 - 9
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broad range of activity
6 - 10
Pseudomonas carrageenovora
-
about 45% of maximal activity
7.4 - 8.4
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about 60% of maximal activity at pH 7.4 and at pH 8.4
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
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assay at room temperature
25 - 50
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30
Tamlana sp.
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39
Zobellia sp.
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-
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 45
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15C: about 40% of maximal activity, 45C: about 50% of maximal activity
20 - 60
25 - 60
Tamlana sp.
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30 - 55
Zobellia sp.
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high activity within this range
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.1
sequence calculation
8.5
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isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32940
-
electrospray mass spectrometry
85000
recombinant enzyme, gel filtration
128000
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gel filtrration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
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1 * 128000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
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processed upon secretion from a 44000 Da preproprotein to a 32000 Da secretion product, first by N-terminal removal of the 25-amino-acid signal peptide, then by cleavage of a 96-amino-acid C-terminal region from amino acids 302-397
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme expressed in Escherichia coli
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seleno-methionyl derivative of protein
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
1 h, about 65% loss of activity
208838
4.5
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1 h, about 40% loss of activity
208838
5
Zobellia sp.
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purified recombinant enzyme, 20C, 2 h, 65% activity remaining
731197
6
-
1 h, about 30% loss of activity
208838
6 - 9
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the activity of the enzyme is stable from pH 6.0 to 9.0, greater than 70.0% of the residual activity is maintained after treatment at pH from 6.0 to 9.0 and 4C for 6 h, 100% of activity is maintained after treatment at pH 8.0 and 4C for 6 h
716808
6 - 8
Zobellia sp.
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purified recombinant enzyme, 20C, 2 h, 85% activity remaining
731197
6.6 - 8.6
-
-
715775
7
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1 h, stable
208838
7.2 - 8.6
Tamlana sp.
-
the enzyme is stable over the range of pH 7.2-8.6 below 45C
714755
8.5
-
1 h, about 50% loss of activity
208838
10
Zobellia sp.
-
purified recombinant enzyme, 20C, 2 h, inactivation
731197
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0 - 50
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the purified kappa-carrageenase is stable in the range of 0-30C and the residual enzyme activity is still retained at 96.1% of the control after treatment at 30C for 1 h. The enzyme is inactivated rapidly at temperatures higher than 30C and is almost inactivated at 50C within 1 h
5
Pseudomonas carrageenovora
-
stable for 2 weeks
20
-
40% loss of activity within 5 days, complete loss of activity within 12 days
28 - 75
-
the enzyme is stable at 28C, but 95% of the activity is lost at 50-75C for 30 min
30
-
2 h, stable
35
Zobellia sp.
-
purified recombinant enzyme, pH 6.0, 3 h, 95% activity remaining
37
-
5 days, 50% loss of activity
45 - 60
Tamlana sp.
-
enzyme activity is stable (more than 91%) for 2 h at temperatures of above 45C. However, 90% of the activity is lost after incubation at 50-60C for 30 min
45
Zobellia sp.
-
purified recombinant enzyme, pH 6.0, 15 min, 20% activity remaining
55
-
10 min, complete inactivation
60
-
purified enzyme, pH 7.0, inactivation within 60 min
70
-
purified enzyme, pH 7.0, inactivation within 30 min
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
repeated freezing and thawing drastically reduces activity
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, stable for at least 16 months
-
-20C, stable for several months
-
20C, 5 days, 40% loss of activity
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4C, 0.2 M Tris-HCl buffer
-
frozen, stable for 15 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, CM-cellulose 52 column chromatography, and Sephadex G-200 gel filtration
-
ammonium sulfate precipitation, Sephadex G-200 gel filtration, and DE-cellulose 52 anion-exchange chromatography
Tamlana sp.
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by metal-affinity chromatography followed by cation exchange chromatography
-
DEAE Sepharose column chromatography and HiTrap column chromatography
-
native enzyme from culture supernatant by ammonium sulfate fractionation and adsorption chromatography
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native extracellular enzyme to electrophoretic homogeneity from culture supernatant by ammonium sulfate fractionation, dialysis, and two different steps of gel filtration
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recombinant CBM-tagged chimeric fusion enzyme cCgkA-cCglA by cellulose affinity purification using cellulose as a support or by nickel affinity chromatography
recombinant enzyme expressed in Escherichia coli
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recombinant extracelluar enzyme 388.6fold from Escherichia coli strain DH5alpha by ammonium sulfate fractionation, dialysis, anion and cation exchange chromatography to homogeneity
recombinant extracellular (24fold) and intracellular (19fold) His-tagged enzyme from Escherichia coli strain BL21 (DE3) cell-free extract and culture supernatant by ammonium sulfate fractionation, dialysis, nickel affinity chromatography and dialysis
Zobellia sp.
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recombinant His-tagged enzyme from the periplasm of Escherichia coli strain BL21(DE3) by nickel affinity chromatography and cation exchange chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, recombinant expression in Escherichia coli strain DH5alpha as extracellular enzyme
expression in Escherichia coli
-
gene cgkA, recombinant expression of functional CBM-tagged chimeric fusion enzyme cCgkA-cCglA, consisting of the catalytic domains of kappa-carrageenase CgkA and lambda-carrageenase CglA from Pseudoalteromonas carrageenovora fused with a dockerin domain, in Escherichia coli strain BL21(DE3), cloning in Escherichia coli strain DH5alpha
gene cgkZ, DNA and amino acid sequence determination and analysis, phylogenetic tree, recombinant expression of His-tagged enzyme with and without signal peptide extracellulary and intracellularly in Escherichia coli strain BL21 (DE3), cloning in Escherichia coli strain DH5alpha
Zobellia sp.
-
gene cgkZ, recombinant expression of N-terminally His6-tagged extracellular enzyme in Escherichia coli strain BL21(DE3), 51% of the total kappa-carrageenase is secreted into culture medium, and 33% accumulates in the periplasmic space. Low temperature of 23C and optimal isopropyl-beta-L-thiogalactoside concentration of 0.9 mM favor soluble protein expression, additives such as lactose, glycine, Tween-80, and Triton X-100 promote the release of intracellular enzymes, method optimization and evaluation, overview
Zobellia sp.
-
His-tagged enzyme expression in the periplasm of Escherichia coli strain BL21(DE3)
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overexpressed as a His-tagged fusion protein in Escherichia coli
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
construction of a chimeric fusion protein consisting of the catalytic domains of kappa-carrageenase CgkA and lambda-carrageenase CglA from Pseudoalteromonas carrageenovora fused with a dockerin domain from Clostridium cellulovorans. The recombinant cCgkA and cCglA are assembled with scaffoldin miniCbpA from Clostridium cellulovorans via cohesin and dockerin interaction. Carbohydrate binding module in scaffoldin is used as a tag
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
Pseudomonas carrageenovora
-
studies of carrageenan structure
molecular biology
Pseudomonas carrageenovora
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potential use in the future in the field of seaweed biotechnology for obtaining protoplasts, and in molecular biology to prevent severe separation problems occuring in the presence of phycocolloid oligomers
pharmacology
Pseudomonas carrageenovora
-
potential use in the future: production of defined phycocolloid oligomers for pharmacy and immunology
additional information