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 hide
3.2.1.89
-
RECOMMENDED NAME
GeneOntology No.
arabinogalactan endo-beta-1,4-galactanase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
The enzyme specifically hydrolyses (1->4)-beta-D-galactosidic linkages in type I arabinogalactans.
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
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-
-
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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.
CAS REGISTRY NUMBER
COMMENTARY hide
58182-40-4
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain KF-267, gene galB
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
strains S-2 and S-39
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-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Fusarium roseum
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene Pcgal1
UniProt
Manually annotated by BRENDA team
gene Pcgal1
UniProt
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
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
-
-
?
arabinogalactan + H2O
galactose
show the reaction diagram
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
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
beta-1,4-galactan + H2O
galactobiose + galactotriose
show the reaction diagram
debranched sugar beet arabinan + H2O
?
show the reaction diagram
galactan + H2O
?
show the reaction diagram
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
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-
?
galactan I + H2O
?
show the reaction diagram
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isolated from black liquor, enzyme produces a range of galacto-oligolaccharides, main products are galactose and galactobiose
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-
?
galactobiose + H2O
galactose
show the reaction diagram
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by galactanase I
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-
?
galactotetraose + H2O
galactose
show the reaction diagram
-
by galactanase I
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-
?
lupin beta-1,4-galactan + H2O
?
show the reaction diagram
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
-
-
-
-
?
soybean arabinogalactan + 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
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
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-
?
additional information
?
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
arabinose
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o-nitrophenyl-beta-D-galactoside hydrolyzing activity
EDTA
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complete inactivation
galactose
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o-nitrophenyl-beta-D-galactoside hydrolyzing activity
glucose
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o-nitrophenyl-beta-D-galactoside hydrolyzing activity
lactose
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o-nitrophenyl-beta-D-galactoside hydrolyzing activity
xylose
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o-nitrophenyl-beta-D-galactoside hydrolyzing activity
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.6
purified recombinant enzyme, substrate lupin galactan, pH 5.0, 37C
166
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galactanase I
180
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galactanase III
199
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galactanase II
224
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galactanase II
238
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galactanase I
3059
-
purified enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 5.5
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at least 50% activity
3.4 - 5
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enzyme acts efficiently between pH 3.4 and pH 5; high activity within this range
4 - 8
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inactive below and above
5 - 8
pH 5.0: about 60% of maximal activity, pH 8.0: about 70% of maximal activity
5.5 - 8.5
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at least 50% activity
6.5 - 9.5
-
at least 50% activity
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 48
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loses activity above, inactive at 60C
70 - 100
70C: about 50% of maximal activity, 100C: about 95% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
below, isoelectric focusing; pI below 4, chromatofocusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
commercial pectinase preparation
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Bacillus licheniformis (strain ATCC 14580 / DSM 13 / JCM 2505 / NBRC 12200 / NCIMB 9375 / NRRL NRS-1264 / Gibson 46)
Bacillus licheniformis (strain ATCC 14580 / DSM 13 / JCM 2505 / NBRC 12200 / NCIMB 9375 / NRRL NRS-1264 / Gibson 46)
Bacillus licheniformis (strain ATCC 14580 / DSM 13 / JCM 2505 / NBRC 12200 / NCIMB 9375 / NRRL NRS-1264 / Gibson 46)
Bacillus licheniformis (strain ATCC 14580 / DSM 13 / JCM 2505 / NBRC 12200 / NCIMB 9375 / NRRL NRS-1264 / Gibson 46)
Bacillus licheniformis (strain ATCC 14580 / DSM 13 / JCM 2505 / NBRC 12200 / NCIMB 9375 / NRRL NRS-1264 / Gibson 46)
Bacillus licheniformis (strain ATCC 14580 / DSM 13 / JCM 2505 / NBRC 12200 / NCIMB 9375 / NRRL NRS-1264 / Gibson 46)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thielavia heterothallica
Thielavia heterothallica
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22000 - 24000
-
gel filtration
37000
-
sedimentation equilibrium
43000
-
calculation from sequence of DNA
48500
-
SDS-PAGE
66000 - 67000
Fusarium roseum
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
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N-glycosylation, mannose residues
proteolytic modification
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additional information
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no glycoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method
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hanging drops, vapour-diffusion, 30% polyethylene glycol 400, 0.2 M CaCl2, 0.1 M Na HEPES, pH 7.5, orthorhombic space group
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presence of divalent cations, preferably Ca2+
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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
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
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concentrated enzyme solution crystallized in the cold
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hanging drop vapor diffusion method
galactanase I forms a hexagonal crystal
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2 - 8
-
-
136947
2.5 - 5.5
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at least 50% activity after 2 h at 37C
657295
3 - 9
purified recombinant enzyme, over 80% activity remaining after 16 h
731199
3
60C, 60 min, about 75% loss of activity
678791
3.2 - 7.5
-
enzyme retains at least 80% of its activity between pH 4.4 and pH 7 at 40C after 24 h, but loses 50% of its activity below pH 3.2 and above pH 7.5; purified enzyme, 24 h, 40C, 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
655166
3.5 - 8.5
-
at least 50% activity after 2 h at 37C
657295
4 - 7
-
-
136949
4 - 10
-
-
136936
4 - 9
60C, 60 min, less than 20% los of activity in the pH-range 4-9
678791
5 - 12
-
S-39
136950
5 - 9.5
-
at 30C
136943
5.5 - 10.5
-
in presence of Ca2+
136942
6 - 10
-
-
136951
7 - 12
-
S-2
136950
10
60C, 60 min, about 60% of maximal activity
678791
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
above 30C the activity is reduced after 30 min
55
-
stable for 10 min at pH 5.5
70
-
stable below
80
pH 7.0, 60 min, no loss of activity
85
pH 7.0, 60 min, about 30% loss of activity
90
pH 7.0, 60 min, about 95% loss of activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C loss of activity
-
4C, 50 mM sodium acetate, pH 5.0, several weeks
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Purification/COMMENTARY
ORGANISM
UNIPROT
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
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native extracellular enzyme from the culture supernatant by ammonium sulfate fractionation, ultrafiltration, anion exchange and hydrophobic interaction chromatography, and gel filtration
partial
recombinant enzyme from Aspergillus oryzae in a one-sep anion exchange chromatography to homogeneity
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recombinant enzyme from Bacillus subtilis by adsorption chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in potato
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expression in Saccharomyces cerevisiae
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gene galB, overexpression in Aspergillus oryzae, optimization of the expression level
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gene Pcgal1, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression in Escherichia coli strain BL21, subcloning in Escherichia coli strain DH5alpha
overexpression in Aspergillus oryzae
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overexpression in Bacillus subtilis
overexpression in Escherichia coli
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D128N
-
increased pH optimum
E263A
-
inactive nucleophile mutant
A90S/H91D
-
decreased pH optimum
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