Information on EC 3.2.1.89 - arabinogalactan endo-beta-1,4-galactanase

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

EC NUMBER
COMMENTARY
3.2.1.89
-
RECOMMENDED NAME
GeneOntology No.
arabinogalactan endo-beta-1,4-galactanase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
The enzyme specifically hydrolyses (1->4)-beta-D-galactosidic linkages in type I arabinogalactans.
show the reaction diagram
-
-
-
-
The enzyme specifically hydrolyses (1->4)-beta-D-galactosidic linkages in type I arabinogalactans.
show the reaction diagram
active site and substrate binding structures, overview
-, Q65CX5
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of O-glycosyl bond
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
arabinogalactan 4-beta-D-galactanohydrolase
This enzyme, isolated from the bacterium Bacillus subtilis, hydrolyses the beta(1->4) bonds found in type I plant arabinogalactans, which are a component of the primary cell walls of dicots. The predominant product is a tetrasaccharide. cf. EC 3.2.1.181, galactan endo-beta-1,3-galactanase.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
arabinogalactanase
-
-
-
-
beta-1,4-galactanase
-
-
beta-D-galactanase
-
-
endo-1,4-beta-galactanase
-
-
-
-
endo-beta-1,4-D-galactanase
-
-
endo-beta-1,4-D-galactanase
Q9X0S8
-
endo-beta-1,4-galactanase
-
-
endo-beta-D-1,4-galactanase
-
-
endo-beta-D-1,4-galactanase
Aspergillus niger KF-267
-
-
-
endogalactanase A
-
-
galactanase
-
-
-
-
GALB
Aspergillus niger KF-267
-
-
-
additional information
-
the enzyme belongs to the glycoside hydrolase family 53, GH-53
CAS REGISTRY NUMBER
COMMENTARY
58182-40-4
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
; from a commercial product Pectinex Ultra SP-L
-
-
Manually annotated by BRENDA team
strain KF-267, gene galB
-
-
Manually annotated by BRENDA team
Aspergillus niger KF-267
strain KF-267, gene galB
-
-
Manually annotated by BRENDA team
strains S-2 and S-39
-
-
Manually annotated by BRENDA team
Fusarium roseum
-
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
arabinogalactan + H2O
galactose
show the reaction diagram
-
-
-
-
?
arabinogalactan + H2O
galactose
show the reaction diagram
-
-
-
-
?
arabinogalactan + H2O
galactose
show the reaction diagram
-
-
-
-
?
arabinogalactan + H2O
galactose
show the reaction diagram
Fusarium roseum
-
-
-
-
?
arabinogalactan + H2O
galactose
show the reaction diagram
-
-
-
-
?
arabinogalactan + H2O
galactose
show the reaction diagram
Bacillus subtilis F-11
-
-
-
-
?
arabinogalactan + H2O
D-galactose
show the reaction diagram
-
initial release of D-galactotriose and D-galactotetraose, prolonged incubation results in D-galactose and D-galactobiose
-
-
?
AZCL-galactan + H2O
?
show the reaction diagram
-
the enzyme shows a conserved beta-turn, characteristic of GH53 enzymes, which contributes to subsites -2 to +3
-
-
?
azurine cross-linked galactan + H2O
?
show the reaction diagram
-
enzyme releases blue color
-
-
?
azurine cross-linked lupin-galactan + H2O
?
show the reaction diagram
-
enzyme releases blue color
-
-
?
beta-(1->4)-D-galactotriose + H2O
D-galactose
show the reaction diagram
-
by galactanase I
-
-
?
beta-1,4-galactan
?
show the reaction diagram
-
main products are galactose and galactobiose
-
-
-
beta-1,4-galactan + H2O
?
show the reaction diagram
-
enzyme produces a range of galacto-oligolaccharides, main products are galactose and galactobiose
-
-
?
beta-1,4-galactan + H2O
?
show the reaction diagram
-
enzyme releases sugars from galactan
-
-
?
beta-1,4-galactan + H2O
?
show the reaction diagram
-
enzyme releases sugars from lupin-galactan
-
-
?
beta-1,4-galactan + H2O
galactobiose + galactotriose
show the reaction diagram
-, Q65CX5
plant-cell-wall-degrading enzyme involved in the hydrolysis of beta-1,4-galactan in the hairy regions of pectin
-
-
?
beta-1,4-galactan + H2O
galactobiose + galactotriose
show the reaction diagram
-, Q65CX5
1% w/v galactan from lupin treated with arabinofuranosidase
product identification
-
?
galactan + H2O
?
show the reaction diagram
Q9X0S8, -
initially, rather long oligosaccharides are detected as hydrolysis products of potato galactan. Then, as the reaction proceeds, mainly galactotetraose is produced. After 24 h reaction, D-galactose, beta-1,4-D-galactobiose, beta-1,4-D-galactotriose and beta-1,4-D-galactotetraose are detected as the major products from potato galactan. The enzyme hydrolyzes galactans more efficiently than pectic galactans
-
-
?
galactan I + H2O
?
show the reaction diagram
-
isolated from black liquor, enzyme produces a range of galacto-oligolaccharides, main products are galactose and galactobiose
-
-
?
galactobiose + H2O
galactose
show the reaction diagram
-
by galactanase I
-
-
?
o-nitrophenyl-beta-D-galactopyranoside + H2O
o-nitrophenol + galactose + pyranose
show the reaction diagram
-
-
-
-
?
p-nitrophenyl-beta-D-galactoside + H2O
p-nitrophenol + galactose
show the reaction diagram
-
-
-
-
?
galactotetraose + H2O
galactose
show the reaction diagram
-
by galactanase I
-
-
?
additional information
?
-
-
involved in pectin degradation
-
-
-
additional information
?
-
-
role in degradation of both linear and L-arabinose-substitued galactan side chains of pectin
-
-
-
additional information
?
-
-
enzyme liberates galactose from a lignin-carbohydrate complex isolated from softwood kraft pulp, no activity towards beta-1,3-linked galactan
-
-
-
additional information
?
-
-
no activity towards D-galacto-oligosaccharides that are substituted with D-glucose at the reducing end
-
-
-
additional information
?
-
-, Q65CX5
no activity with galactobiose and galactotriose as substrates
-
-
-
additional information
?
-
-
activity with galactans from different sources, overview, the enzyme is highly active and specific for beta-galactans hydrolyzing beta-1,4-galactosidic linkages, substrate specificity, the enzyme acts on beta-1,4-linked galactan, producing a range of galacto-oligosaccharides, and on beta-1,4- and beta-1,3/6-linked arabinogalactans, the enzyme liberates galactose from a lignin–carbohydrate complex isolated from softwood kraft pulp, no activity towards beta-1,3-liked galactan, overview
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
arabinogalactan + H2O
galactose
show the reaction diagram
-
-
-
-
?
beta-1,4-galactan + H2O
galactobiose + galactotriose
show the reaction diagram
-, Q65CX5
plant-cell-wall-degrading enzyme involved in the hydrolysis of beta-1,4-galactan in the hairy regions of pectin
-
-
?
additional information
?
-
-
involved in pectin degradation
-
-
-
additional information
?
-
-
role in degradation of both linear and L-arabinose-substitued galactan side chains of pectin
-
-
-
additional information
?
-
-
activity with galactans from different sources, overview
-
-
-
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
arabinose
-
o-nitrophenyl-beta-D-galactoside hydrolyzing activity
EDTA
-
complete inactivation
galactose
-
o-nitrophenyl-beta-D-galactoside hydrolyzing activity
glucose
-
o-nitrophenyl-beta-D-galactoside hydrolyzing activity
lactose
-
o-nitrophenyl-beta-D-galactoside hydrolyzing activity
xylose
-
o-nitrophenyl-beta-D-galactoside hydrolyzing activity
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
21.8
-
Q9X0S8, -
-
166
-
-
galactanase I
180
-
-
galactanase III
199
-
-
galactanase II
224
-
-
galactanase II
238
-
-
galactanase I
3059
-
-
purified enzyme
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3.5
-
-
wild-type enzyme
4
-
-
galactanase I
4
-
-
S-39
4.5
-
-
galactanase II
4.5
-
-
D128N mutant
5
-
Fusarium roseum
-
-
6.5
-
-
A90S/H91D mutant
7
-
Q9X0S8, -
-
7.5
-
-
wild-type enzyme
7.5
-
-, Q65CX5
-
10
-
-
S-2
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3
5.5
-
at least 50% activity
3.4
5
-
enzyme acts efficiently between pH 3.4 and pH 5; high activity within this range
4
8
-
inactive below and above
5
8
Q9X0S8, -
pH 5.0: about 60% of maximal activity, pH 8.0: about 70% of maximal activity
5.5
8.5
-
at least 50% activity
6.5
9.5
-
at least 50% activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
at pH 6.5
40
-
-
S-39
40
-
-, Q65CX5
-
50
-
-
S-2
50
-
-
assay at
50
-
-
at pH 3.5
90
-
Q9X0S8, -
-
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
48
-
loses activity above, inactive at 60°C
70
100
Q9X0S8, -
70°C: about 50% of maximal activity, 100°C: about 95% of maximal activity
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
-
-
below, isoelectric focusing; pI below 4, chromatofocusing
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
commercial pectinase preparation
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Bacillus licheniformis (strain DSM 13 / ATCC 14580)
Bacillus licheniformis (strain DSM 13 / ATCC 14580)
Bacillus licheniformis (strain DSM 13 / ATCC 14580)
Bacillus licheniformis (strain DSM 13 / ATCC 14580)
Bacillus licheniformis (strain DSM 13 / ATCC 14580)
Bacillus licheniformis (strain DSM 13 / ATCC 14580)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Thielavia heterothallica
Thielavia heterothallica
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
22000
24000
-
gel filtration
32000
-
-
galactanase III, gel filtration
35000
-
-
galactanase I, gel filtration
36000
-
-
galactanase II, gel filtration
36000
-
-
S-2
37000
-
-
sedimentation equilibrium
38000
-
-
Western blotting
38000
-
-
SDS-PAGE
38000
-
-
gel filtration; SDS-PAGE, gel filtration
43000
-
-
calculation from sequence of DNA
44000
-
-
recombinant enzyme
48500
-
-
SDS-PAGE
66000
67000
Fusarium roseum
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 42000, recombinant enzyme, SDS-PAGE
?
Q9X0S8, -
x * 64000, SDS-PAGE
?
-
x * 44000, SDS-PAGE of mutant E263A
?
Aspergillus niger KF-267
-
x * 42000, recombinant enzyme, SDS-PAGE
-
monomer
-
1 * 35000 SDS-PAGE
monomer
-
1 * 39700 SDS-PAGE
monomer
-
1 * 38000, SDS-PAGE
additional information
-, Q65CX5
secondary and three-dimensional structure modelling and analysis, the enzyme structure shows a(betaalpha)8-barrel architecture and a calcium ion linking the long betaalpha-loops 7 and 8
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
-
-
glycoprotein
-
N-glycosylation, mannose residues
additional information
-
no glycoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
hanging drop vapor diffusion method
-
hanging drops, vapour-diffusion, 30% polyethylene glycol 400, 0.2 M CaCl2, 0.1 M Na HEPES, pH 7.5, orthorhombic space group
-
presence of divalent cations, preferably Ca2+
-
purified enzyme free or in complex with products galactobiose and galactotriose, hanging drop vapour diffusion method, 0.002-0.004 ml of protein solution, containing 20-40 mg/ml protein, are mixed with 0.002 ml reservoir solution containing 21-26% w/v PEG 1500, X-ray diffraction structure determination and analysis at 2.6 A resolution, molecular replacement modelling
-, Q65CX5
wild-type and mutant E263A in complex with methyl-beta(1-4)-galactotetraoside, grown at room temperature in hanging drops with a resolution range of 30-2.3 A. Crystals of wild-type and mutant E263A both belong to the monoclinic space group P21, containing two molecules in the asymmetric unit
-
concentrated enzyme solution crystallized in the cold
-
hanging drop vapor diffusion method
-
galactanase I forms a hexagonal crystal
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2.5
5.5
-
at least 50% activity after 2 h at 37°C
3
-
Q9X0S8, -
60°C, 60 min, about 75% loss of activity
3.2
7.5
-
enzyme retains at least 80% of its activity between pH 4.4 and pH 7 at 40°C after 24 h, but loses 50% of its activity below pH 3.2 and above pH 7.5; purified enzyme, 24 h, 40°C, 80% remaining activity at pH 4.4 and pH 7.0, less than 50% remaining activity below pH 3.2 and above pH 7.5
3.5
8.5
-
at least 50% activity after 2 h at 37°C
4
9
Q9X0S8, -
60°C, 60 min, less than 20% los of activity in the pH-range 4-9
5
12
-
S-39
5
9.5
-
at 30°C
5.5
10.5
-
in presence of Ca2+
7
12
-
S-2
10
-
Q9X0S8, -
60°C, 60 min, about 60% of maximal activity
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
-
-
above 30°C the activity is reduced after 30 min
40
-
-
up to, purified enzyme, pH 4.5, 24 h, 80% remaining activity
45
-
-
S-2, at pH 10 for 15 min stable
45
-
-
purified enzyme, pH 4.5, 24 h, 47% remaining activity
50
-
-
stable below
50
-
-
stable below
55
-
-
stable for 10 min at pH 5.5
60
-
-
stable below
60
-
-
S-39 at pH 4 for 15 min stable
60
-
-
complete loss of activity
70
-
-
stable below
80
-
Q9X0S8, -
pH 7.0, 60 min, no loss of activity
85
-
Q9X0S8, -
pH 7.0, 60 min, about 30% loss of activity
90
-
Q9X0S8, -
pH 7.0, 60 min, about 95% loss of activity
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20°C loss of activity
-
4°C, 50 mM sodium acetate, pH 5.0, several weeks
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
from a commercial product Pectinex Ultra SP-L; native enzyme from commercial pectinase preparation by anion and cation exchange chromatography, ultrafiltration, and gel filtration
-
recombinant enzyme from Aspergillus oryzae in a one-sep anion exchange chromatography to homogeneity
-
recombinant enzyme from Bacillus subtilis by adsorption chromatography
-, Q65CX5
recombinant
Q9X0S8, -
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in potato
-
expression in Saccharomyces cerevisiae
-
overexpression in Aspergillus oryzae
-
gene galB, overexpression in Aspergillus oryzae, optimization of the expression level
-
overexpression in Bacillus subtilis
-, Q65CX5
overexpression in Escherichia coli
Q9X0S8, -
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D128N
-
increased pH optimum
E263A
-
inactive nucleophile mutant
A90S/H91D
-
decreased pH optimum