Information on EC 3.2.1.163 - 1,6-alpha-D-mannosidase

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

EC NUMBER
COMMENTARY hide
3.2.1.163
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RECOMMENDED NAME
GeneOntology No.
1,6-alpha-D-mannosidase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of the (1->6)-linked alpha-D-mannose residues in alpha-D-Manp-(1->6)-D-Manp
show the reaction diagram
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-
-
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SYSTEMATIC NAME
IUBMB Comments
(1->6)-alpha-mannosyl alpha-D-mannohydrolase
The enzyme is specific for (1->6)-linked mannobiose and has no activity towards any other linkages, or towards p-nitrophenyl-alpha-D-mannopyranoside or baker's yeast mannan. It is strongly inhibited by Mn2+ but does not require Ca2+ or any other metal cofactor for activity.
CAS REGISTRY NUMBER
COMMENTARY hide
90910-03-5
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
physiological function
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the enzyme is the pneumococcal virulence factor in Streptococcus pneumoniae
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-methylumbelliferyl alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranoside + H2O
4-methylumbelliferone + alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranose
show the reaction diagram
4-nitrophenyl alpha-D-mannopyranoside + H2O
4-nitrophenol + alpha-D-mannopyranose
show the reaction diagram
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Epman is highly specific in its cleavage of the alpha-1,6 mannosidic linkage
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-
?
alpha-D-mannopyranosyl-(1,6)-D-mannose + H2O
?
show the reaction diagram
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-
-
-
?
alpha-D-Manp-(1,6)-D-Manp + H2O
alpha-D-mannose
show the reaction diagram
Man-alpha(1-3)[Man-alpha(l-6)]Man-beta(1-4)GlcNAc-beta(1-4)GlcNAc-beta(1-N)Asn + H2O
?
show the reaction diagram
-
i.e. Man3GlcNAc2Asn, degradation in concert with endoglycosidases, low activity, overview
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-
?
Man-alpha(1-6)Man-alpha(1-4)GlcNAc-beta(1-4)GlcNAc-beta(1-N)Asn + H2O
?
show the reaction diagram
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i.e. Man2GlcNAc2Asn, degradation in concert with endoglycosidases, low activity, overview
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-
?
additional information
?
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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
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
-
Epman is stimulated 1.5fold with 1 mM Mg2+
Mn2+
-
Epman is stimulated 1.25fold with 1 mM Mn2+
additional information
-
Ca2+ is not required for activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2R,3R,4S)-2-([[(1R)-2-hydroxy-1-phenylethyl]amino]methyl)pyrrolidine-3,4-diol
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(3R,4R,5R)-3,4-dihydroxy-5-([[(1R)-2-hydroxy-1-phenylethyl]amino]methyl)-1-methylpyrrolidin-2-one
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(3R,4R,5R)-3,4-dihydroxy-5-([[(1R)-2-hydroxy-1-phenylethyl]amino]methyl)pyrrolidin-2-one
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1,4-Dideoxy-1,4-imino-D-mannitol
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Mannostatin A
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Mn2+
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strong inhibition
swainsonine
additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.38
4-methylumbelliferyl alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranoside
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pH 7.4, 37C
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2 - 3
4-nitrophenyl alpha-D-mannopyranoside
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in 100 mM sodium acetate at pH 4.0 and 37C
14
alpha-D-Manp-(1-6)-D-Manp
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pH 4.0, 60C
additional information
additional information
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kinetics
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4.9
4-methylumbelliferyl alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranoside
Clostridium perfringens
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pH 7.4, 37C
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
12.9
4-methylumbelliferyl alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranoside
Clostridium perfringens
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pH 7.4, 37C
202178
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.255
(2R,3R,4S)-2-([[(1R)-2-hydroxy-1-phenylethyl]amino]methyl)pyrrolidine-3,4-diol
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in 100 mM sodium acetate (pH 4.0), at 37C
0.026
(3R,4R,5R)-3,4-dihydroxy-5-([[(1R)-2-hydroxy-1-phenylethyl]amino]methyl)-1-methylpyrrolidin-2-one
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in 100 mM sodium acetate (pH 4.0), at 37C
0.21
(3R,4R,5R)-3,4-dihydroxy-5-([[(1R)-2-hydroxy-1-phenylethyl]amino]methyl)pyrrolidin-2-one
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in 100 mM sodium acetate (pH 4.0), at 37C
0.0006
Mannostatin A
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in 100 mM sodium acetate at pH 4.0 and 37C
0.004
swainsonine
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in 100 mM sodium acetate at pH 4.0 and 37C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0049
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culture supernatant, at pH 4.0
0.9
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after 182fold purification, at pH 4.0
48
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purified enzyme
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 5
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36% of maximal activity at pH 3.0, rapid inactivation above pH 5.0
3.6 - 5.5
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45 - 70
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21% of maximal activity at 45C, 16% at 70C
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.6
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isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
-
the enzyme is expressed in all tissues
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
74000
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1 * 74000, SDS-PAGE
81000
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gel filtration
120000
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SDS-PAGE
123600
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MALDI-TOF mass spectrometry
180000
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recombinant enzyme, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
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1 * 74000, SDS-PAGE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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purified enzyme, stable for 24 h, rapid inactivation above
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
freeze and thaw cycles and freeze drying have a detrimental effect on the purified enzyme with loss of 60% activity
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
23C, room temperature, purified enzyme, over 95% remaining activity after 7 days
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4C, purified enzyme, over 95% remaining activity after 3 months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Cibacron blue F3GA resin column chromatography, Sepharose column chromatography, and Superdex 200 gel filtration
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native enzyme 370fold by anion-exchange chromatography, chromatofocussing, and gel filtration
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native enzyme partially by affinity chromatography
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recombinant His- or HA-tagged enzyme
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression as His- or HA-tagged protein
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expressed in Drosophila S2 cells
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recombinant expression in Pichia pastoris
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis