Information on EC 3.2.1.152 - mannosylglycoprotein endo-beta-mannosidase

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The expected taxonomic range for this enzyme is: Magnoliophyta

EC NUMBER
COMMENTARY hide
3.2.1.152
-
RECOMMENDED NAME
GeneOntology No.
mannosylglycoprotein endo-beta-mannosidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of the alpha-D-mannosyl-(1->6)-beta-D-mannosyl-(1->4)-N-acetyl-beta-D-glucosaminyl-(1->4)-N-acetyl-beta-D-glucosaminyl sequence of glycoprotein to alpha-D-mannosyl-(1->6)-D-mannose and N-acetyl-beta-D-glucosaminyl-(1->4)-N-acetyl-beta-D-glucosaminyl sequences
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
CAS REGISTRY NUMBER
COMMENTARY hide
141176-95-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
Manually annotated by BRENDA team
-
-
-
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
(Man)nMan-alpha-(1-6)-Man-beta-(1-4)GlcNAc-beta-(1-4)GlcNAc-peptide + pyridylamino-GlcNAc-beta-(1-4)GlcNAc
pyridylamino-(Man)nMan-alpha-(1-6)-Man-beta-(1-4)GlcNAc-beta-(1-4)GlcNAc + GlcNAc-beta-(1-4)GlcNAc-peptide
show the reaction diagram
-
n = 0-2
-
-
?
Man-alpha(1-6)[Man-alpha(1-3)]Man-alpha-(1-6)Man-beta-(1-4)GlcNAc-beta-(1-4)GlcNAc-pyridylamide + H2O
Man-alpha(1-6)[Man-alpha(1-3)]Man-alpha-(1-6)Man + GlcNAc-beta-(1-4)GlcNAc-pyridylamide
show the reaction diagram
-
-
-
-
?
Man-alpha-(1-3)Man-alpha-(1-6)Man-beta-(1-4)GlcNAc-beta-(1-4)GlcNAc-pyridylamine + H2O
GlcNAc-beta-(1-4)GlcNAc-pyridylamine + Man-alpha-(1-3)Man-alpha-(1-6)Man
show the reaction diagram
-
relative hydrolysis rate: 42%
-
-
?
Man-alpha-(1-6)(Man-alpha-(1-3))Man-alpha-(1-6)Man-beta-(1-4)GlcNAc-beta-(1-4)GlcNAc-pyridylamine + H2O
GlcNAc-beta-(1-4)GlcNAc-pyridylamine + Man-alpha-(1-6)(Man-alpha-(1-3))Man-alpha-(1-6)Man
show the reaction diagram
-
relative hydrolysis rate: 34%
-
-
?
Man-alpha-(1-6)Man-alpha(1-6)Man-beta-(1-4)GlcNAc-beta-(1-4)GlcNAc-pyridylamide + H2O
Man-alpha-(1-6)Man-alpha(1-6)Man + GlcNAc-beta-(1-4)GlcNAc-pyridylamide
show the reaction diagram
-
-
-
-
?
Man-alpha-(1-6)Man-beta-(1-4)GlcNAc-beta-(1-4)GlcNAc-pyridylamide + H2O
Man-alpha-(1-6)Man + GlcNAc-beta-(1-4)GlcNAc-pyridylamide
show the reaction diagram
-
-
-
-
?
Man-alpha-(1-6)Man-beta-(1-4)GlcNAc-beta-(1-4)GlcNAc-pyridylamine + H2O
GlcNAc-beta-(1-4)GlcNAc-pyridylamine + Man-alpha-(1-6)Man
show the reaction diagram
Man-beta-(1-4)GlcNAc + H2O
?
show the reaction diagram
Man-beta-(1-4)GlcNAc-beta-(1-4)GlcNAc-peptide + p-nitrophenyl-beta-D-glucoside
Man-beta-(1-4)GlcNAc-beta-(1-4)Glc-beta-1-p-nitrophenyl + GlcNAc-peptide
show the reaction diagram
-
-
-
-
?
Man-beta-(1-4)GlcNAc-beta-(1-4)GlcNAc-peptide + p-nitrophenyl-beta-D-mannoside
Man-beta-(1-4)GlcNAc-beta-1-p-nitrophenyl-beta-D-mannoside + GlcNAc-peptide
show the reaction diagram
-
-
-
-
?
Man-beta-(1-4)GlcNAc-beta-(1-4)GlcNAc-peptide + p-nitrophenyl-beta-N-acetylglucosaminide
Man-beta-(1-4)GlcNAc-beta-1-p-nitrophenyl-beta-N-acetylglucosaminide + GlcNAc-peptide
show the reaction diagram
-
-
-
-
?
Man-beta-(1->4)-GlcNAc-beta-(1->4)-GlcNAc-beta-pyridylamine + H2O
D-mannose + GlcNAc-beta-(1->4)-GlcNAc-beta-pyridylamine
show the reaction diagram
-
relative hydrolysis rate: 9%
-
-
?
Man-beta-1-4GlcNAc + H2O
?
show the reaction diagram
-
hydrolysis of Man-beta-1-4GlcNAc linkage of N-linked sugar chains
-
-
ir
Manalpha(1-3)[Manalpha(1-6)]Manalpha(1-6)Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-pyridylaminoside
Manalpha(1-3)[Manalpha(1-6)]Manalpha(1-6)Man + GlcNAcbeta(1-4)GlcNAc-pyridylaminoside
show the reaction diagram
-
40% activity relative to M2B-PA
-
-
?
Manalpha(1-3)[Manalpha(1-6)]Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-pyridylaminoside + H2O
Manalpha(1-3)[Manalpha(1-6)]Man + GlcNAcbeta(1-4)GlcNAc-pyridylaminoside
show the reaction diagram
-
34% activity relative to Manalpha(1-6)Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-pyridylaminoside
-
-
?
Manalpha(1-6)Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-pyridylaminoside + H2O
Manalpha(1-6)Man + GlcNAcbeta(1-4)GlcNAc-pyridylaminoside
show the reaction diagram
-
100% activity
-
-
?
Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-pyridylaminoside + H2O
beta-D-mannose + GlcNAcbeta(1-4)GlcNAc-pyridylaminoside
show the reaction diagram
-
2.6% activity relative to M2B-PA
-
-
?
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
Man-beta-(1-4)GlcNAc + H2O
?
show the reaction diagram
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.2 - 1.4
Man-alpha-(1-6)Man-beta-(1-4)GlcNAc-beta-(1-4)GlcNAc-pyridylamine
0.54
Manalpha(1-6)Manbeta(1-4)GlcNAcbeta(1-4)GlcNAc-pyridylaminoside
-
in ammonium acetate buffer, pH 5.0, at 37C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.004
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purified enzyme, pH 5.0, 37C
0.026
purified enzyme, pH 5.0, 37C
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
78000
-
gel filtration
108000
calculated from the deduced amino acid sequence
141000
-
SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
three bands on SDS-PAGE of 42000, 31000, and 28000 Da
tetramer
-
1 * 42000 + 1 * 34000 + 1 * 28000 + 1 * 10000, gel filtration
trimer
alphabetagamma, 1 * 42000 + 1 * 31000 + 1 * 28000, SDS-PAGE
additional information
-
SDS-PAGE of the purified protein shows that EBM II is comprised of six polypeptides with molecular masses of 65000, 46000, 42000, 31000, 28000 and 27000 Da. Analysis of the fragments disclose EBM II consists of endo-beta-mannosidase and an associated protein that has slight homology to the fucosidase domain of the bifidus AfcA protein
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
enzyme is encoded by a single cDNA, translation product is posttranslationally processed by proteolysis
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 7
unstable below pH 3 and above pH 7
659383
3 - 6
-
relatively stable at pH 3-6
669154
3 - 8
unstable below pH 3 and above pH 8
659098
additional information
-
unstable at alkaline pH
659075
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; ammonium sulfate precipitation, Poros HS column chromatography, hydroxyapatite chromatography, DEAE-Sephacel column chromatography, Octyl Sepharose CL-4B column chromatography, and Superdex 200 gel filtration
-
enzyme is purified by ammonium sulfate precipitation, DEAE-Sephacel chromatography, sephacryl S-200 gel filtration, octyl sepharose CL-4B chromatography and superdex 200 gel filtration
-
partial purification of the recombinant protein
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli RosettaBlue
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E464A
no activity
E549A
no activity
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
-
possible application in the synthesis of oligosaccharides containing mannosyl-beta-1,4-structures