Information on EC 3.2.1.81 - beta-agarase

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

EC NUMBER
COMMENTARY hide
3.2.1.81
-
RECOMMENDED NAME
GeneOntology No.
beta-agarase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of (1->4)-beta-D-galactosidic linkages in agarose, giving the tetramer as the predominant product
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
agarose degradation
-
-
porphyran degradation
-
-
SYSTEMATIC NAME
IUBMB Comments
agarose 4-glycanohydrolase
Also acts on porphyran, but more slowly [1]. This enzyme cleaves the beta-(1->4) linkages of agarose in a random manner with retention of the anomeric-bond configuration, producing beta-anomers that give rise progressively to alpha-anomers when mutarotation takes place [6]. The end products of hydrolysis are neoagarotetraose and neoagarohexaose in the case of AgaA from the marine bacterium Zobellia galactanivorans, and neoagarotetraose and neoagarobiose in the case of AgaB [6].
CAS REGISTRY NUMBER
COMMENTARY hide
37288-57-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
; strain AG LSL-1, enzyme is inducible by agar and not repressed by other simple sugars. NaCl is not required for growth or production of agarase. Enzyme belongs to group III beta-agarase family
-
-
Manually annotated by BRENDA team
strain AG LSL-1, enzyme is inducible by agar and not repressed by other simple sugars. NaCl is not required for growth or production of agarase. Enzyme belongs to group III beta-agarase family
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Agarivorans sp.
strain JAMB-A11
-
-
Manually annotated by BRENDA team
strain C-1
-
-
Manually annotated by BRENDA team
E-1
-
-
Manually annotated by BRENDA team
MK03
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Cellvibrio sp.
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Microbulbifer sp.
Microbulbifer sp. CMC-5 (MTCC 9889)
-
-
-
Manually annotated by BRENDA team
Microbulbifer sp. JAMB-A94
strain JAMB-A94
UniProt
Manually annotated by BRENDA team
strain JAMB-A94
UniProt
Manually annotated by BRENDA team
strain N-1
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain SK38
-
-
Manually annotated by BRENDA team
Pseudomonas-like bacterium
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
strain F-6
Uniprot
Manually annotated by BRENDA team
strain JT0107
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
strain V134
UniProt
Manually annotated by BRENDA team
strain V143
-
-
Manually annotated by BRENDA team
Dsij
UniProt
Manually annotated by BRENDA team
strain KCTC 12921
-
-
Manually annotated by BRENDA team
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
agar + H2O
neoagarobiose + neoagarotetraose + neoagarohexaose
show the reaction diagram
agar + H2O
neoagarohexaose + neoagarotetraose
show the reaction diagram
-
-
-
-
?
agar + H2O
neoagarooctaose + neoagarohexaose + neoagarotetraose
show the reaction diagram
-
-
-
?
agar + H2O
neoagarotetraose + neoagarohexaose
show the reaction diagram
agaropectin + H2O
?
show the reaction diagram
-
-
-
-
ir
agarose + H2O
2 neoagarooligosaccharides
show the reaction diagram
rAgaC with low activity (0.05 U/ml) cleaves agarose into longer neoagarooligosaccharides. When activity is increased to 30 U/ml, neoagarooctaose is produced predominantly in addition to neoagarohexaose and neoagarotetraose
-
-
?
agarose + H2O
?
show the reaction diagram
agarose + H2O
agarotetraose
show the reaction diagram
agarose + H2O
neoagarobiose
show the reaction diagram
agarose + H2O
neoagarobiose + neoagarotetraose
show the reaction diagram
agarose + H2O
neoagarobiose + neoagarotetraose + neoagarohexaose
show the reaction diagram
agarose + H2O
neoagarohexaose + neoagarotetraose
show the reaction diagram
agarose + H2O
neoagarohexaose + neoagarotetraose + neoagarobiose
show the reaction diagram
agarose + H2O
neoagarooctaose + ?
show the reaction diagram
agarose + H2O
neoagarooctaose + neoagarodecaose
show the reaction diagram
agarose + H2O
neoagarosebiose
show the reaction diagram
agarose + H2O
neoagarotetraose
show the reaction diagram
agarose + H2O
neoagarotetraose + ?
show the reaction diagram
agarose + H2O
neoagarotetraose + neoagarobiose
show the reaction diagram
agarose + H2O
neoagarotetraose + neoagarohexaose
show the reaction diagram
agarose + H2O
neoagarotetraose + neoagarohexaose + neoagarobiose
show the reaction diagram
agarose + H2O
neoagarotetraose + neoagarohexaose + neoagarooctaose
show the reaction diagram
alginate + H2O
?
show the reaction diagram
Microbulbifer sp.
-
-
-
-
?
alkali treated porphyran + H2O
?
show the reaction diagram
carboxymethyl cellulose + H2O
?
show the reaction diagram
carrageenan + H2O
?
show the reaction diagram
Microbulbifer sp.
-
-
-
-
?
chitin + H2O
?
show the reaction diagram
neoagaro-oligosaccharides + H2O
?
show the reaction diagram
-
from N2 to N22, reaction yield for neoagaro-oligosaccharides is 52.7% by 4 U/mg beta-agarase
-
-
?
neoagarodecaose + H2O
?
show the reaction diagram
is a very poor substrate
-
-
?
neoagarododecaose + H2O
?
show the reaction diagram
-
-
-
?
neoagarododecaose + H2O
neoagarohexaose + neoagarotetraose + neoagarooctaose
show the reaction diagram
-
-
-
?
neoagarohexaitol + H2O
?
show the reaction diagram
-
-
-
-
ir
neoagarohexaose + H2O
?
show the reaction diagram
neoagarohexaose + H2O
neoagarobiose
show the reaction diagram
neoagarohexaose + H2O
neoagarobiose + neoagarotetraose
show the reaction diagram
neoagarohexaose + H2O
neoagarotetraose + neoagarobiose
show the reaction diagram
neoagarooctaose + H2O
neoagarobiose + neoagarotetraose + neoagarohexaose
show the reaction diagram
-
-
-
?
neoagarooctaose + H2O
neoagarohexaose + neoagarobiose
show the reaction diagram
neoagarooctaose + H2O
neoagarohexaose + neoagarotetraose + neoagarobiose
show the reaction diagram
-
-
-
-
?
neoagarooctaose + H2O
neoagarotetraose
show the reaction diagram
neoagarotetradecaose + H2O
?
show the reaction diagram
-
-
-
?
neoagarotetraose + H2O
neoagarobiose
show the reaction diagram
polysaccharides with neoagarobiose units + H2O
neoagarobiose + neoagarotetraose
show the reaction diagram
-
-
-
-
ir
porphyran + H2O
neoagaro-oligosaccharides
show the reaction diagram
porphyran + H2O
neoagarohexaose + neoagarotetraose
show the reaction diagram
-
-
-
-
?
porphyran + H2O
neoagarooctaose + neoagarohexaose + neoagarotetraose
show the reaction diagram
-
-
-
?
xylan + 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
agar + H2O
neoagarobiose + neoagarotetraose + neoagarohexaose
show the reaction diagram
agar + H2O
neoagarotetraose + neoagarohexaose
show the reaction diagram
agarose + H2O
neoagarobiose
show the reaction diagram
agarose + H2O
neoagarobiose + neoagarotetraose + neoagarohexaose
show the reaction diagram
agarose + H2O
neoagarohexaose + neoagarotetraose + neoagarobiose
show the reaction diagram
agarose + H2O
neoagarooctaose + ?
show the reaction diagram
agarose + H2O
neoagarooctaose + neoagarodecaose
show the reaction diagram
-
-
major products
-
?
agarose + H2O
neoagarotetraose
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
135% activity at 1 mM
Cu2+
1 mM, increases activity of agarase AG-b 1.2fold
Fe2+
2 mM stimulates agarase activity by 20%
KCl
-
no activity in the presence of 00.3 M salt. The enzyme shows a salt requirement for activity, being active from 0.3 M NaCl, with maximal activity at 3.5 M NaCl. KCl supports similar activities as NaCl up to 3.5 M, and LiCl up to 2.5 M. The monovalent salts can not be substituted by 3.5 M divalent cations, CaCl2 or MgCl2
LiCl
-
no activity in the presence of 00.3 M salt. The enzyme shows a salt requirement for activity, being active from 0.3 M NaCl, with maximal activity at 3.5 M NaCl. KCl supports similar activities as NaCl up to 3.5 M, and LiCl up to 2.5 M. The monovalent salts can not be substituted by 3.5 M divalent cations, CaCl2 or MgCl2
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-ethyl-(3-(3-dimethyl-amino-)propyl)carbonate
Microbulbifer sp.
-
1 mM. 10-20% inhibition
5,5'-dithiobis-(2-nitrobenzoic acid)
53% residual activity at 10 mM
Ag+
5 mM, complete inhibition of agarase AG-b
Ba2+
Microbulbifer sp.
-
85% residual activity at 1 mM
Co2+
Microbulbifer sp.
-
complete inhibition at 1 mM
diethyl dicarbonate
Microbulbifer sp.
-
1 mM, 10-20% inhibition
EGTA
-
the inhibition by 1 mM EGTA is almost completely recovered by the addition of Co2+ at a concentration of 0.2 mM
FeCl2
-
2 mM, 99% inhibition
FeCl3
-
partially inhibits the activity
K+
-
94.9% residual activity at 100 mM
KCl
-
50% inhibition at 0.1 M
L-galactose 6-sulfate
-
-
MnCl2
-
partially inhibits the activity
N-bromosuccinimide
NH4Cl
-
50% inhibition at 0.1 M
Pb(CH3COO)2
PbCl2
-
2 mM, 98% inhibition
sodium dodecylsulfate
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
30% increase in activity
beta-mercaptoethanol
beta-mercaptoethanol even at a low concentration of 10 mM increases the activity of AgaA by 23%
CaCl2
-
stimulation
dithiothreitol
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0182 - 4.2
agarose
1 - 2
liquid-phase agarose
12.81
neoagarodecaose
-
12.39
neoagarododecaose
-
1.7
neoagarohexaose
-
-
8.36
neoagarotetradecaose
-
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
22 - 41
agarose
0.19
neoagarodecaose
Pseudoalteromonas sp. CY24
A1A3Y9
-
4.77
neoagarododecaose
Pseudoalteromonas sp. CY24
A1A3Y9
-
5.16
neoagarotetradecaose
Pseudoalteromonas sp. CY24
A1A3Y9
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.3
agarose
Saccharophagus degradans
-
in 20 mM Tris-HCl buffer (pH 7.0), at 30C
2642
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1
KCl
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.04
Microbulbifer sp.
-
culture supernatant, pH 7.0, 30C
0.1
-
AgaB, 1 ml of 0.125% melted agarose, 44C, pH 7.5
0.16
-
AgaAc, 1 ml of 0.125% melted agarose, 44C, pH 7.5
0.7
Cellvibrio sp.
-
crude enzyme from culture fluid, in 50 mM phosphate buffer, pH 7.0, 35C
0.872
-
cell lysate
0.9
Agarivorans sp.
cell-free extract, at pH 7.0 and 35C
1.2
-
wild-type
3
-
cell-free medium, at pH 7.0 and 35C
4.13
-
concentrated agarase solution
4.22
Microbulbifer sp.
-
after 103.5fold purification, pH 7.0, 30C
9.3
-
-
14.2
-
pH 7.0, 40C
16.9
culture fluid
25.5
-
40C, pH 8.0
36
mutant enzyme V109G/V110C/T111H/S112L, in 20 mM phosphate buffer (pH 6.5) containing 0.25% (w/v) agarose at 40C for 10 min
57.45
-
agarase Hz-c, 13.9fold purified
63.6
-
catalytic module
76.8
-
agarase Hz-b, 18.6fold purified
83.5
-
35C, pH 7.0; after 27.8fold purification, at pH 7.0 and 35C
84.2
Cellvibrio sp.
-
after 120.2fold purification, in 50 mM phosphate buffer, pH 7.0, 35C
167
Agarivorans sp.
-
pH 8.0, 40C
204.4
using agar as substrate, in pH 5.5 acetate buffer, at 45C
207.5
using agarose as substrate, in pH 5.5 acetate buffer, at 45C
208.1
Agarivorans sp.
purified enzyme, at pH 7.0 and 35C
230.1
agarase AG-a
242.2
15fold purified rAgaB34, with agarose as substrate at 40C
329
-
recombinant AgaC
371
Agarivorans sp.
-
50 mM N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid-NaOH buffer, pH 7.8, containing 10 mM CaCl2, 0.1 mM NaCl, 40C
466
mutant enzyme N103T, in 20 mM phosphate buffer (pH 6.5) containing 0.25% (w/v) agarose at 40C for 10 min
475
mutant enzyme S182I, in 20 mM phosphate buffer (pH 6.5) containing 0.25% (w/v) agarose at 40C for 10 min
482
wild type enzyme, in 20 mM phosphate buffer (pH 6.5) containing 0.25% (w/v) agarose at 40C for 10 min
1600
-
purified refolded AgaB
5000
recombinant AgaB 476fold purified
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2
-
porphyran as substrate
6 - 9
Agarivorans sp.
-
6.5 - 7.5
native enzyme
6.7
Pseudomonas-like bacterium
-
beta-agarase I and IIb
6.8
-
porphyran as substrate
7.4
-
porphyran as substrate
7.5 - 8
Agarivorans sp.
-
Britton-Robinson universal buffers, MOPS and N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid-NaOH buffer
additional information
-
mutant L122Q/N446I has comparable optimum pH as the wild-type
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 9
-
the enzyme shows more than 90% activity between pH 5.0 and 8.0. More than 50% of the agarase activity is lost below pH 4.0 and above pH 9.0
4 - 8
-
-
4.5 - 7.5
the enzyme retains its activity up to 60% at pH 4.5-7.5
5 - 8
-
pH 5.0: about 35% of maximal activity, about 40% of maximal activity
5 - 8.5
5 - 9
-
about 60% of activity
5 - 10
pH 5.0: about 65% of maximal activity, pH 10.0: about 75% of maximal activity, agarase AG-a
5.5 - 7.5
Cellvibrio sp.
-
the agarase exhibits more than 90% of the maximum activity in the pH range of 5.5 to 7.5
5.5 - 8
-
-
6 - 9
-
more than 80% of maximum activity
6 - 8
Agarivorans sp.
the enzyme activity is about 80% at pH 6.0 and less than 60% over pH 8.0
6 - 8
Cellvibrio sp.
more than 50% activity between pH 6.0 and 8.0. The activity gradually decreases at alkaline pH and significantly declines at acidic pH
6 - 8
Microbulbifer sp.
-
the agarase retains more than 70% of its original activity at pH 6.0 and 8.0
6 - 9
-
more than 80% of the maximal activity retained
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 40
34
-
porphyran as substrate
38
Pseudomonas-like bacterium
-
beta-agarase I
38 - 55
native enzyme
40 - 41
-
-
42.5
Cellvibrio sp.
-
43
Pseudomonas-like bacterium
-
beta-agarase IIb
54
Microbulbifer sp.
-
-
60
Agarivorans sp.
-
more than 70% of maximum activity
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20
-
30% of maximum activity
25 - 55
Agarivorans sp.
the enzyme activity is more than 80% at 25C and 45C and less than 10% over 55C
25 - 47
Cellvibrio sp.
more than 50% activity between 25 and 47C. The enzyme activity drastically decreases over 45C
25 - 40
-
the enzyme works efficiently in the range of 25-40C and shows maximum activity at 40C but loses essentially all of its activity at 60C
25 - 70
25C: about 60% of maximal activity, 70C: about 80% of maximal activity, agarase AG-a
30
-
90% of maximum activity
30 - 45
-
-
40 - 50
Cellvibrio sp.
-
the agarase exhibits more than 90% of the maximum activity in the temperature ranging from 40C to 50C
45 - 85
-
45C: about 50% of maximal activity, 85C: about 55% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.1
-
calculated
4.14
-
-
5.4
calculated from amino acid sequence
5.9
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Saccharophagus degradans (strain 2-40 / ATCC 43961 / DSM 17024)
Saccharophagus degradans (strain 2-40 / ATCC 43961 / DSM 17024)
Saccharophagus degradans (strain 2-40 / ATCC 43961 / DSM 17024)
Saccharophagus degradans (strain 2-40 / ATCC 43961 / DSM 17024)
Zobellia galactanivorans (strain DSM 12802 / CIP 106680 / NCIMB 13871 / Dsij)
Zobellia galactanivorans (strain DSM 12802 / CIP 106680 / NCIMB 13871 / Dsij)
Zobellia galactanivorans (strain DSM 12802 / CIP 106680 / NCIMB 13871 / Dsij)
Zobellia galactanivorans (strain DSM 12802 / CIP 106680 / NCIMB 13871 / Dsij)
Zobellia galactanivorans (strain DSM 12802 / CIP 106680 / NCIMB 13871 / Dsij)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
13400
-
AgaAc, calculated by sequence
20000
-
SDS-PAGE
21000
-
SDS-PAGE
26500
-
gel filtration
28000
-
SDS-PAGE
29000
-
calculation from sequence of DNA
31000
recombinant enzyme
32150
-
AgaAc, ESI MS
35130
-
calculation from sequence of an open reading frame
36000
-
SDS-PAGE
37000
recombinant enzyme
40010
-
AgaB, ESI MS
47240
-
sequence analysis
48200
Microbulbifer sp.
-
calculated from sequence
48400
calculated from amino acid sequence
50900
sequence analysis
51200
calculated from amino acid sequence
56000
-
gel filtration
58000
Agarivorans sp.
-
60000
recombinant enzyme
61000
Pseudomonas-like bacterium
-
gel filtration, beta-agarase IIb
63000
-
SDS-PAGE
63600
Pseudomonas-like bacterium
-
analytical ultracentrifugation, beta-agarase IIb
65020
Microbulbifer sp.
-
calculated from sequence
72000
-
SDS-PAGE
72500
maltose binding protein-fused enzyme, SDS-PAGE
79000
Cellvibrio sp.
gel filtration
82000
-
SDS-PAGE
84000
-
His-tagged recombinant enzyme, SDS-PAGE
87500
native enzyme
89000
recombinant enzyme
100000
105000
106100
-
sequence analysis
109000
Agarivorans sp.
recombinant enzyme
113000
127000
Pseudomonas-like bacterium
-
gel filtration, beta-agarase I
140000
Pseudomonas-like bacterium
-
SDS-PAGE, beta-agarase I
146000
recombinant enzyme
180000
-
gel filtration
210000
Pseudomonas-like bacterium
-
analytical ultracentrifugation, beta-agarase I
353000
-
AgaB, calculated by sequence
539000
-
AgaA, calculated by sequence
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
Pseudomonas-like bacterium
-
galactose, N-acetylglucosamine and glucosamine
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
sitting drop vapor diffusion method, using 0.1 M HEPES pH 7.0, 30% PEG 4000, 0.1 M ammonium sulfate and 15% glycerol
-
carbohydrate binding module CBM6-2 of isoform Aga16B in complex with neoagarohexaose. The carbohydrate binding module targets the equatorial O4 and the axial O3 of the anhydro-L-galactose moiety
-
E147S mutant in complex with agaro-octaose, 1.7 A resolution, structure determination by molecular replacment, hanging drop vapor duffusion method, 20.5C, 30% PEG 4000, 200 mM ammonium acetate, 100 mM sodium acetate
hanging drop vapor diffusion method, using 28-34% (w/v) PEG 8000 and 0.1 M imidazole pH 8.0, at 19C
inactive mutant A-E174S, in complex with agaro-octaose
-
recombinant protein expressed in Escherichia coli
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 11
4 - 10
4C, 6 h, stable in the pH range of 4.010.0, agarase AG-a
699362
4 - 9
-
12 h, 25C, retains more than 80% of activity
678815
4 - 8
-
705484
5 - 9
-
699497
5 - 8
Microbulbifer sp.
-
705484
5.5 - 11
5.7 - 10.6
6 - 11
-
-
677787
6 - 9
-
705484
6 - 11
Agarivorans sp.
-
retaining 70% of original activity
668031
7 - 8.6
the enzyme is most stable from pH 7.0 to 8.6
713851
7.1 - 8.2
8
-
the enzyme is stable at pH 8.0 after 30 min incubation
713704
8 - 9
-
705484
additional information
-
mutant L122Q/N446I has comparable pH stability as the wild-type
695671
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 30
the enzyme retains its activity when the temperature was between 4 and 30C for 1 h
20 - 40
Cellvibrio sp.
the enzyme is stable from 20 to 40C after 30 min of incubatiopn. At 40C, the enzyme retains 90% activity, however, it drastically decreases by 22% of the original value at 42.5C
20 - 45
-
heat treatment in the range of 20-40C does not affect the residual activity of the enzyme. Only 45% of the activity is retained after heating at 50C for 1 h
25 - 45
Agarivorans sp.
the enzyme activity is stable at 25C and 35C. The enzyme retains more than 80% activity after 2 h exposure at the indicated temperature. However, it maintains less than 40% activity after 1 h at 45C and 55C
25
Agarivorans sp.
stable up to 25C
47.5
-
loss in activity 53%
50 - 90
-
the agarase is heat-labile, with rapid loss of activity when treated at 50C for 15 min or at 70C for 1 min. It loses above 80% of its original activity after incubation at 90C for 20 s
50 - 60
Microbulbifer sp.
-
the agarase is thermally stable up to 50C, with 62% of its residual activity is retained and 45% of the agarase activity is still retained at 60C. The agarase activity is completely abolished at 70C
50 - 60
-
mutant enzymes E99K, T307I and E99K/T307I are stable up to 50C, the melting temperature of E99K/T307I is increased by 5.2C over that of the wild type enzyme (54.6C). The E99K/T307I mutant enzyme is stable at 55C with 1 mM CaCl2, reaching 260% of the activity the wild type enzyme held at 40C without CaCl2
65 - 80
-
the enzyme retains approximately 90% of the initial activity after incubation for 1 h at 65-80C
70
-
1 min, 20% residual activity
95
-
the enzyme retains 50% activity after 1 h at 95C
100
-
in the presence of 10 mM CaCl2, approximately 17% remaining activity is detected after 30 min
additional information
-
mutant L122Q/N446I has higher thermostability than wild-type AgaB. Melting temperature of S2 is 4.6C higher than that of wild-type AgaB, and the half-life of mutant L122Q/N446I is 350 min at 40C, which is 18.4fold longer than that of the wild-type enzyme. Mutant N446I has similar enhanced thermostability as mutant L122Q/N446I. L122Q shows similar thermostability with wild-type AgaB. Half-life of mutant N446V at 40C is 18.2fold longer than that of wild-type AgaB. Half-life of mutant N446L at 40C is 12.9fold longer than that of wild-type AgaB
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
maintains more than half of its maximum activity at an NaCl concentration of 4 M
-
salt requirement for stability
-
the agarase shows almost the same activity in both the 0 mM and 1000 mM NaCl solutions, and even
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 20 mM Tris-HCl, more than 12 months
-
-20C, at least 6 months
Pseudomonas-like bacterium
-
-20C, more than 2 years
-
4C, 20 mM Tris-HCl buffer, more than 12 months
-
4C, pH 7.5 (phosphate buffer), 6 months, agarase AG-a loses 8% activity
4C, Tris-HCl with 0.4 M NaCl, pH 7.5, more than 6 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
17.9fold with a final yield of 23.3%
-
18.8fold and with a final yield of 16.8% by anion exchange and hydrophobic chromatographies
-
476fold and with a final yield of 21.2%, by ammonium sulfate precipitation, weak anion-exchange chromatography, hydrophobic interaction chromatography, and gel filtration
; ammonium sulfate precipitation, DEAE-Sepharose column chromatography, and Sephacryl S-100HR gel filtration; ammonium sulfate precipitation, gel filtration on Sephacryl S-100HR, ion exchange chromatography on diethylaminoethyl-Sepharose
-
agarose bead chromatography
-
ammonium sulfate precipitation
-
ammonium sulfate precipitation and Sephadex gel filtration
-
ammonium sulfate precipitation, DEAE-Toyopearl PAK-650M column chromatography, and Sephacryl S-200 gel filtration
Cellvibrio sp.
ammonium sulfate precipitation, phenyl Sepharose column chromatography, and Superdex 75 gel filtration
ammonium sulfate precipitation, Sepharose CL6B, anion-exchange Mono Q HR5, Ni-NTA column, AgaB further gel filtration with Sephacryl S200
-
ammonium sulfate precipitation, Toyopearl QAE-550C column chromatography, Toyopearl HW-55F column chromatography, and Mono-Q column chromatography
Cellvibrio sp.
-
chitin bead column chromatography
from culture supernatant
-
His-trap HP Ni2+ column chromatography and Sephacryl S-200 gel filtration
HisTrap column chromatography
-
isolated from the sediment in Suruga bay, Japan, at a depth of 2406m, purification by ammonium sulfate precipitation, DEAE-Toyopearl 650M, gel filtration
Microbulbifer sp.
-
isolated from the sediment of south side of the Kuril trench, Japan, at a depth of 4152 m, purification by ammonium sulfate precipitation, dialysis, DEAE-Toyopearl 650M column, dialysis
Agarivorans sp.
-
Ni affinity column chromatography and Sephacryl S-200 gel filtration
Ni-NTa affinity chromatography and exclusion chromatography with Sephacryl S100
Ni-NTA column chromatography
Ni2+-Sepharose column chromatography
-
nickel-affinity column chromatography, HiTrap DEAE column chromatography, and Superdex 200 gel filtration
-
rAgaB34 purified by ultrafiltration and on Ni+ column, 15fold with a yield of 80%
recombinant AgaB and mutants purified by ammonium sulfate precipitation, weak anion-exchange chromatography, hydrophobic interaction chromatography, and gel filtration. Mutant S2 purified about 476.5fold, with a final yield of 20.9%
-
recombinant protein isolated from culture broth
Agarivorans sp.
-
to electrophoretic purity by immobilized metal affinity chromatography
-
to homogeneity directly from the native-PAGE without stained by Coomassie brilliant blue, agarase Hz-b 18.6fold purified and agarase Hz-c 13.9fold purified
-
ultrafiltration, DEAE Sepharose column chromatography and Sephacryl S-300HR gel filtration
Microbulbifer sp.
-
using the pMAL protein fusion and purification system