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EC Tree
IUBMB Comments 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.
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Reaction Schemes
Hydrolysis of the (1->6)-linked alpha-D-mannose residues in alpha-D-Manp-(1->6)-D-Manp
Synonyms
man2b2, 1,6-alpha-d-mannosidase, alpha-1,6-mannosidase, core-specific alpha1,6-mannosidase, lysosomal alpha (1-6)-mannosidase, alpha-1,6-linkage-specific mannosidase,
more
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alpha 1,6-mannosidase
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alpha-1,6-linkage-specific mannosidase
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alpha-1,6-mannosidase
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alpha-1,6-specific mannosidase
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core-specific alpha1,6-mannosidase
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core-specific lysosomal alpha (1-6)-mannosidase
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core-specific lysosomal alpha 1,6-mannosidase
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GH125 exo-alpha-1,6-mannosidase
lysosomal alpha (1-6)-mannosidase
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lysosomal alpha 1,6-mannosidase
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additional information
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the enzyme belongs to the glycosylhydrolase family 38
GH125 exo-alpha-1,6-mannosidase
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GH125 exo-alpha-1,6-mannosidase
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Hydrolysis of the (1->6)-linked alpha-D-mannose residues in alpha-D-Manp-(1->6)-D-Manp
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(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.
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4-methylumbelliferyl alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranoside + H2O
4-methylumbelliferone + alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranose
4-nitrophenyl alpha-D-mannopyranoside + H2O
4-nitrophenol + alpha-D-mannopyranose
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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
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alpha-D-Manp-(1,6)-D-Manp + H2O
alpha-D-mannose
Man-alpha(1-3)[Man-alpha(l-6)]Man-beta(1-4)GlcNAc-beta(1-4)GlcNAc-beta(1-N)Asn + H2O
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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
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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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4-methylumbelliferyl alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranoside + H2O
4-methylumbelliferone + alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranose
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synthesis of the fluorogenic disaccharide substrate and assay method development, overview
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4-methylumbelliferyl alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranoside + H2O
4-methylumbelliferone + alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranose
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synthesis of the fluorogenic disaccharide substrate and assay method development, overview
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alpha-D-Manp-(1,6)-D-Manp + H2O
alpha-D-mannose
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the enzyme cleaves specifically (1-6)-linked mannobiose and has no activity towards alpha-D-Manp-1,2-D-Manp or alpha-D-Manp-1, 3-D-Manp
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alpha-D-Manp-(1,6)-D-Manp + H2O
alpha-D-mannose
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the enzyme cleaves specifically (1-6)-linked mannobiose and has no activity towards any other linkages
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additional information
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no activity with p-nitrophenyl-alpha-D-mannopyranoside or baker's yeast mannan
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additional information
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regioselective synthesis of mannobiose and mannotriose by reverse hydrolysis of commercially available manno-oligosaccharide
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additional information
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the core-specific lysosomal enzyme is involved in N-glycan catabolism and degradation of Man3GlcNAc2 N-glycan structures, requiring for chitobiase action before alpha1,6-mannosidase activity
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additional information
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the enzyme is active on the branched trimannosyl core of complex glycans cleaving alphal-6-linked mannose residues from branched Man3Glc-NAc to form Manalphal-3Manbeta1-4GlcNAc, overview, activities of alpha-mannosidases in fibroblasts, overview
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additional information
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substrate specificity, the enzyme cleaves alphal-6-linked mannose residues from branched Man3Glc-NAc to form Manalphal-3Manbeta1-4GlcNAc, but shows little activity on artificial substrates 4-methylumbelliferyl alpha-mannoside and 4-nitrophenyl alpha-mannoside, overview
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additional information
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the core-specific enzyme is specific for cleaving only the alpha1-6mannose residue from Man3GlcNAc but not Man3GlcNAc2 or other larger high mannose oligosaccharides, NMR analysis of substrates and products, overview
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additional information
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the core-specific lysosomal enzyme is involved in N-glycan catabolism, Man-alpha(l-6)linkage hydrolysis depends on the action of endoglycosidases, namely aspartyl-N-acetyl-beta-D-glucosaminidase and endo-N-acetyl-beta-D-glucosaminidase, overview
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additional information
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the enzyme does not possess the ability to decrease the extent of the O-mannose chains on glycoprotein A expressed in a GFI1.0 glycoengineered Pichia pastoris strain nor the ability to remove the single O-linked mannose residue
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additional information
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the recombinant enzyme shows no activity with recombinant human granulocyte colony-stimulating factor protein (rhGCSF) expressed in a GFI2.0 glycoengineered Pichia pastoris strain and possessing O-linked glycosylation composed of a single O-linked mannose residue. alpha-1,6-Mannosidase from Xanthomonas manihotis does not possess the ability to decrease the extent of the O-mannose chains
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additional information
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additional information
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the core-specific lysosomal enzyme is involved in N-glycan catabolism and degradation of Man3GlcNAc2 N-glycan structures, requiring for chitobiase action before alpha1,6-mannosidase activity
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additional information
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the enzyme is active on the branched trimannosyl core of complex glycans cleaving alphal-6-linked mannose residues from branched Man3Glc-NAc to form Manalphal-3Manbeta1-4GlcNAc, overview, activities of alpha-mannosidases in fibroblasts, overview
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additional information
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the core-specific lysosomal enzyme is involved in N-glycan catabolism, Man-alpha(l-6)linkage hydrolysis depends on the action of endoglycosidases, namely aspartyl-N-acetyl-beta-D-glucosaminidase and endo-N-acetyl-beta-D-glucosaminidase, overview
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Mg2+
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Epman is stimulated 1.5fold with 1 mM Mg2+
Mn2+
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Epman is stimulated 1.25fold with 1 mM Mn2+
additional information
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Ca2+ is not required for activity
Co2+
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stimulates
Co2+
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Epman is activated 3fold with 1 mM Co2+
Zn2+
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stimulates
Zn2+
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contains Zn2+ in the active site, Epman is stimulated more than 1.5fold with 1 mM Zn2+
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(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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swainsonine
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additional information
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no inhibition by EDTA at 10 mM
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additional information
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Epman is not affected by 1 mM EDTA
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Carcinoma, Hepatocellular
Circular RNA MAN2B2 promotes cell proliferation of hepatocellular carcinoma cells via the miRNA-217/MAPK1 axis.
Cardiomegaly
Cardiomyocyte Proteome Remodeling due to Isoproterenol-Induced Cardiac Hypertrophy during the Compensated Phase.
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0.38
4-methylumbelliferyl alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranoside
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pH 7.4, 37°C
2 - 3
4-nitrophenyl alpha-D-mannopyranoside
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in 100 mM sodium acetate at pH 4.0 and 37°C
14
alpha-D-Manp-(1-6)-D-Manp
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pH 4.0, 60°C
additional information
additional information
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kinetics
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4.9
4-methylumbelliferyl alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranoside
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pH 7.4, 37°C
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12.9
4-methylumbelliferyl alpha-D-mannopyranosyl-(1->6)-beta-D-mannopyranoside
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pH 7.4, 37°C
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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 37°C
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 37°C
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 37°C
0.0006
Mannostatin A
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in 100 mM sodium acetate at pH 4.0 and 37°C
0.004
swainsonine
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in 100 mM sodium acetate at pH 4.0 and 37°C
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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
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4
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5
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assay at
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3 - 5
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36% of maximal activity at pH 3.0, rapid inactivation above pH 5.0
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37
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assay at
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45 - 70
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21% of maximal activity at 45°C, 16% at 70°C
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4.6
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isoelectric focusing
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brenda
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ATCC 14332
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ATCC 14332, syn. Aspergillus saitoi
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brenda
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brenda
core-specific alpha1,6-mannosidase
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brenda
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brenda
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brenda
additional information
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the enzyme is expressed in all tissues
brenda
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brenda
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brenda
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physiological function
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the enzyme is the pneumococcal virulence factor in Streptococcus pneumoniae
evolution
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the enzyme belongs to the glycoside hydrolase family 125, GH125
evolution
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the enzyme belongs to the glycoside hydrolase family 125, GH125
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123600
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MALDI-TOF mass spectrometry
180000
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recombinant enzyme, gel filtration
74000
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1 * 74000, SDS-PAGE
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monomer
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1 * 74000, SDS-PAGE
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30
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purified enzyme, stable for 24 h, rapid inactivation above
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freeze and thaw cycles and freeze drying have a detrimental effect on the purified enzyme with loss of 60% activity
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23°C, room temperature, purified enzyme, over 95% remaining activity after 7 days
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4°C, purified enzyme, over 95% remaining activity after 3 months
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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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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 Escherichia coli
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recombinant expression in Pichia pastoris
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synthesis
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regioselective synthesis of mannobiose and mannotriose by reverse hydrolysis using the 1,6-alpha-D-mannosidase from Aspergillus phoenicis, method optimization, overview
synthesis
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synthesis of FimH receptor-active manno-oligosaccharides by reverse hydrolysis using alpha-mannosidases from Penicillium citrinum, Aspergillus phoenicis and almond in a sequential reaction process, method development and optimization, overview
synthesis
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the organism is employed in regioselective synthesis of manno-oligosaccharides involving the enzyme
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Haeuw, J.F.; Grard, T.; Alonso, C.; Strecker, G.; Michalski, J.C.
The core-specific lysosomal alpha(1-6)-mannosidase activity depends on aspartamidohydrolase activity
Biochem. J.
297
463-466
1994
Rattus norvegicus
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brenda
Athanasopoulos, V.I.; Niranjan, K.; Rastall, R.A.
The production, purification and characterization of two novel alpha-D-mannosidases from Aspergillus phoenicis
Carbohydr. Res.
340
609-617
2005
Aspergillus phoenicis
brenda
Maitin, V.; Athanasopoulos, V.; Rastall, R.A.
Synthesis of FimH receptor-active manno-oligosaccharides by reverse hydrolysis using alpha-mannosidases from Penicillium citrinum, Aspergillus phoenicis and almond
Appl. Microbiol. Biotechnol.
63
666-671
2004
Aspergillus phoenicis
brenda
Daniel, P.F.; Evans, J.E.; De Gasperi, R.; Winchester, B.; Warren, C.D.
A human lysosomal alpha (1-6)-mannosidase active on the branched trimannosyl core of complex glycans
Glycobiology
2
327-336
1992
Homo sapiens
brenda
Park, C.; Meng, L.; Stanton, L.H.; Collins, R.E.; Mast, S.W.; Yi, X.; Strachan, H.; Moremen, K.W.
Characterization of a human core-specific lysosomal alpha 1,6-mannosidase involved in N-glycan catabolism
J. Biol. Chem.
280
37204-37216
2005
Homo sapiens
brenda
Athanasopoulos, V.I.; Niranjan, K.; Rastall, R.A.
Regioselective synthesis of mannobiose and mannotriose by reverse hydrolysis using a novel 1,6-alpha-D-mannosidase from Aspergillus phoenicis
J. Mol. Catal. B
27
215-219
2004
Aspergillus phoenicis
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brenda
Venkatesan, M.; Kuntz, D.; Rose, D.
Human lysosomal alpha-mannosidases exhibit different inhibition and metal binding properties
Protein Sci.
18
2242-2251
2009
Homo sapiens
brenda
Gomathinayagam, S.; Hamilton, S.
In vitro enzymatic treatment to remove O-linked mannose from intact glycoproteins
Appl. Microbiol. Biotechnol.
98
2545-2554
2014
Xanthomonas phaseoli pv. manihotis
brenda
Deng, L.; Tsybina, P.; Gregg, K.; Mosi, R.; Zandberg, W.; Boraston, A.; Vocadlo, D.
Synthesis of 4-methylumbelliferyl alpha-D-mannopyranosyl-(1-6)-beta-D-mannopyranoside and development of a coupled fluorescent assay for GH125 exo-alpha-1,6-mannosidases
Bioorg. Med. Chem.
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4839-4845
2013
Clostridium perfringens, Streptococcus pneumoniae
brenda
Gomathinayagam, S.; Hamilton, S.
In vitro enzymatic treatment to remove O-linked mannose from intact glycoproteins
Appl. Microbiol. Biotechnol.
98
2545-2554
2014
Xanthomonas phaseoli pv. manihotis
brenda
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