Information on EC 3.2.1.101 - mannan endo-1,6-alpha-mannosidase

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The expected taxonomic range for this enzyme is: Bacillus circulans

EC NUMBER
COMMENTARY hide
3.2.1.101
-
RECOMMENDED NAME
GeneOntology No.
mannan endo-1,6-alpha-mannosidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Random hydrolysis of (1->6)-alpha-D-mannosidic linkages in unbranched (1->6)-mannans
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
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-
-
-
SYSTEMATIC NAME
IUBMB Comments
6-alpha-D-mannan mannanohydrolase
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CAS REGISTRY NUMBER
COMMENTARY hide
58500-56-4
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98148-86-8
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
alpha-1,6-D-mannan + H2O
D-mannose + alpha-1,6-D-mannobiose
show the reaction diagram
alpha-1,6-D-mannohexaose + H2O
D-mannose + alpha-1,6-D-mannobiose
show the reaction diagram
-
-
end-product, with intermediate formation of mannose, mannobiose and mannotriose, the latter apparently converts more slowly to the smaller products
?
alpha-1,6-D-mannooctaose + H2O
D-mannose + alpha-1,6-D-mannobiose
show the reaction diagram
-
largest substrate
end-product, with intermediate formation of mannose, mannobiose and mannotriose, the latter apparently converts more slowly to the smaller products
?
alpha-1,6-D-mannooligosaccharide + H2O
mannose + alpha-1,6-D-mannobiose
show the reaction diagram
alpha-1,6-D-mannopentaose + H2O
D-mannose + alpha-1,6-D-mannobiose
show the reaction diagram
-
-
end-product, with intermediate formation of mannose, mannobiose and mannotriose, the latter apparently converts more slowly to the smaller products
?
alpha-1,6-D-mannotetraose + H2O
D-mannose + alpha-1,6-D-mannobiose
show the reaction diagram
-
-
end-product, with intermediate formation of mannose, mannobiose and mannotriose, the latter apparently converts more slowly to the smaller products
?
alpha-1,6-D-mannotriose + H2O
D-mannose + alpha-1,6-D-mannobiose
show the reaction diagram
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smallest substrate
-
?
alpha-1,6-mannosylated pro-alpha-factor + H2O
?
show the reaction diagram
-
-
-
-
?
reduced mannohexaose + H2O
?
show the reaction diagram
-
poor substrate
-
-
?
reduced mannooctaose + H2O
?
show the reaction diagram
-
poor substrate
-
-
?
reduced mannopentaose + H2O
?
show the reaction diagram
-
poor substrate
-
-
?
reduced mannotetraose + H2O
?
show the reaction diagram
-
poor substrate
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
activation, required for full activity
Co2+
-
very slight activation
Mn2+
-
activation
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
diisopropyl fluorophosphate
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-
EDTA
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70% inhibition, 90% of original activity restored with 10 mM Ca2+
ethanol
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10% ethanol: inactivation
iodoacetamide
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2 mM: 50% inhibition
Mannan
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Saccharomyces cerevisiae wild type mannan, 20% inhibition
Mg2+
-
very slight inhibition
N-ethylmaleimide
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2 mM: 50% inhibition
p-chloromercuribenzoate
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1 mM: complete inhibition, 0.001 mM: 50% inhibition
reduced alpha-1,6-mannohexaose
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54% inhibition, also serves as a substrate
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reduced alpha-1,6-mannotriose
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competitive inhibitor, 92% inhibition
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ethanol
-
1% v/v ethanol: slight activation
iodoacetamide
-
1 mM: slight activation
N-ethylmaleimide
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1 mM: slight activation
Propanol
-
slight activation
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0044
alpha-1,6-D-mannan
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-
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4
alpha-1,6-D-mannobiose
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-
0.072
alpha-1,6-D-mannohexaose
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-
0.12
alpha-1,6-D-mannopentaose
-
-
0.33
alpha-1,6-D-mannotetraose
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1
alpha-1,6-D-mannotriose
-
-
additional information
additional information
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kinetic data
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7.33
alpha-1,6-D-mannan
Bacillus circulans
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-
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2
iodoacetamide
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-
2
N-ethylmaleimide
-
-
0.001
p-chloromercuribenzoate
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482)
Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482)
Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
61000
2 * 61000, SDS-PAGE
125000
-
gel filtration and SDS-PAGE
130000
gel filtration
131000
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sedimentation equilibrium
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
2 * 61000, SDS-PAGE
monomer
-
1 * 125000, SDS-PAGE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
55
-
stable at, in the presence of substrate, for at least 3 h
60 - 65
-
5 min, denatured
additional information
-
unusually heat-stable
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ethanol
-
1% v/v ethanol: stable and slight activation, effect decreases at higher concentrations, at 10% ethanol: inactivation
propanol
-
stable and slight activation
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-10°C, crude enzyme solution, 0.05 M citrate-phosphate buffer, pH 5.5, 0.001 mM CaCl2
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4°C, 0.1 M citrate-phosphate buffer, pH 6.0, or 0.1 M potassium phosphate buffer, pH 6.8, 1 mM CaCl2, stable for at least 1 month
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4°C, 1 ml aliquots, 0.1 M citrate-phosphate buffer, pH 6.0, or 0.1 M potassium phosphate buffer, pH 6.8, 1 mM CaCl2, 1 year, 80% original activity retained
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4°C, crude enzyme solution, 0.1 M citrate-phosphate buffer, pH 5.5, more than 1 month, without appreciable loss of activity
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frozen and lyophilized, crude enzyme solution, more than 1 month, without appreciable loss of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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