BRENDA - Enzyme Database

The production, purification and characterization of two novel alpha-D-mannosidases from Aspergillus phoenicis

Athanasopoulos, V.I.; Niranjan, K.; Rastall, R.A.; Carbohydr. Res. 340, 609-617 (2005)

Data extracted from this reference:

Application
EC Number
Application
Commentary
Organism
3.2.1.163
synthesis
the organism is employed in regioselective synthesis of manno-oligosaccharides involving the enzyme
Aspergillus phoenicis
General Stability
EC Number
General Stability
Organism
3.2.1.163
freeze and thaw cycles and freeze drying have a detrimental effect on the purified enzyme with loss of 60% activity
Aspergillus phoenicis
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.2.1.114
EDTA
10 mM
Aspergillus phoenicis
3.2.1.114
Mn2+
-
Aspergillus phoenicis
3.2.1.163
Mg2+
-
Aspergillus phoenicis
3.2.1.163
Mn2+
strong inhibition
Aspergillus phoenicis
3.2.1.163
additional information
no inhibition by EDTA at 10 mM
Aspergillus phoenicis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.2.1.114
14
-
alpha-D-Man-(1,6)-D-Man
-
Aspergillus phoenicis
3.2.1.163
14
-
alpha-D-Manp-(1-6)-D-Manp
pH 4.0, 60°C
Aspergillus phoenicis
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.2.1.114
Ca2+
1 mM
Aspergillus phoenicis
3.2.1.114
additional information
does not need Ca2+ or any other metal cofactor of those tested
Aspergillus phoenicis
3.2.1.163
additional information
Ca2+ is not required for activity
Aspergillus phoenicis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.2.1.114
74000
-
SDS-PAGE
Aspergillus phoenicis
3.2.1.114
81000
-
native-PAGE
Aspergillus phoenicis
3.2.1.114
97000
-
SDS-PAGE
Aspergillus phoenicis
3.2.1.114
110000
-
native-PAGE
Aspergillus phoenicis
3.2.1.163
74000
-
1 * 74000, SDS-PAGE
Aspergillus phoenicis
3.2.1.163
81000
-
gel filtration
Aspergillus phoenicis
Organism
EC Number
Organism
UniProt
Commentary
Textmining
3.2.1.114
Aspergillus phoenicis
-
-
-
3.2.1.163
Aspergillus phoenicis
-
ATCC 14332, syn. Aspergillus saitoi
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
3.2.1.114
anion-exchange chromatography, chromatofocusing chromatography, gel filtration
Aspergillus phoenicis
3.2.1.163
native enzyme 370fold by anion-exchange chromatography, chromatofocussing, and gel filtration
Aspergillus phoenicis
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.2.1.163
48
-
purified enzyme
Aspergillus phoenicis
Storage Stability
EC Number
Storage Stability
Organism
3.2.1.163
23°C, room temperature, purified enzyme, over 95% remaining activity after 7 days
Aspergillus phoenicis
3.2.1.163
4°C, purified enzyme, over 95% remaining activity after 3 months
Aspergillus phoenicis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
3.2.1.114
alpha-D-Man-(1,6)-D-Man + H2O
-
664527
Aspergillus phoenicis
alpha-D-mannopyranose
-
-
-
?
3.2.1.114
alpha-D-Manp-(1,3)-D-Manp + H2O
-
664527
Aspergillus phoenicis
alpha-D-mannopyranose
-
-
-
?
3.2.1.114
alpha-D-Manp-(1,6)-D-Manp + H2O
-
664527
Aspergillus phoenicis
alpha-D-mannopyranose
-
-
-
?
3.2.1.114
baker's yeast mannan + H2O
-
664527
Aspergillus phoenicis
?
-
-
-
?
3.2.1.114
p-nitrophenyl-alpha-D-mannopyranoside + H2O
-
664527
Aspergillus phoenicis
p-nitrophenol + alpha-D-mannopyranose
-
-
-
?
3.2.1.163
alpha-D-Manp-(1,6)-D-Manp + H2O
the enzyme cleaves specifically (1-6)-linked mannobiose and has no activity towards any other linkages
664527
Aspergillus phoenicis
alpha-D-mannose
-
-
-
?
3.2.1.163
additional information
no activity with p-nitrophenyl-alpha-D-mannopyranoside or baker's yeast mannan
664527
Aspergillus phoenicis
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
3.2.1.163
monomer
1 * 74000, SDS-PAGE
Aspergillus phoenicis
Synonyms
EC Number
Synonyms
Commentary
Organism
3.2.1.114
1,3(1,6)-alpha-D-mannosidase
-
Aspergillus phoenicis
3.2.1.114
1,6-alpha-D-mannosidase
-
Aspergillus phoenicis
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.2.1.114
60
-
-
Aspergillus phoenicis
3.2.1.163
60
-
-
Aspergillus phoenicis
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.2.1.163
30
-
purified enzyme, stable for 24 h, rapid inactivation above
Aspergillus phoenicis
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.2.1.163
4
4.5
-
Aspergillus phoenicis
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.2.1.114
4
4.5
-
Aspergillus phoenicis
3.2.1.163
3
5
36% of maximal activity at pH 3.0, rapid inactivation above pH 5.0
Aspergillus phoenicis
pI Value
EC Number
Organism
Commentary
pI Value Maximum
pI Value
3.2.1.114
Aspergillus phoenicis
for the first isoform of the enzyme, isoelectric focusing
-
4.3
3.2.1.114
Aspergillus phoenicis
method not mentioned
-
4.6
3.2.1.114
Aspergillus phoenicis
for the second isoform of the enzyme, isoelectric focusing
-
4.8
3.2.1.163
Aspergillus phoenicis
isoelectric focusing
-
4.6
Application (protein specific)
EC Number
Application
Commentary
Organism
3.2.1.163
synthesis
the organism is employed in regioselective synthesis of manno-oligosaccharides involving the enzyme
Aspergillus phoenicis
General Stability (protein specific)
EC Number
General Stability
Organism
3.2.1.163
freeze and thaw cycles and freeze drying have a detrimental effect on the purified enzyme with loss of 60% activity
Aspergillus phoenicis
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.2.1.114
EDTA
10 mM
Aspergillus phoenicis
3.2.1.114
Mn2+
-
Aspergillus phoenicis
3.2.1.163
Mg2+
-
Aspergillus phoenicis
3.2.1.163
Mn2+
strong inhibition
Aspergillus phoenicis
3.2.1.163
additional information
no inhibition by EDTA at 10 mM
Aspergillus phoenicis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.2.1.114
14
-
alpha-D-Man-(1,6)-D-Man
-
Aspergillus phoenicis
3.2.1.163
14
-
alpha-D-Manp-(1-6)-D-Manp
pH 4.0, 60°C
Aspergillus phoenicis
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.2.1.114
Ca2+
1 mM
Aspergillus phoenicis
3.2.1.114
additional information
does not need Ca2+ or any other metal cofactor of those tested
Aspergillus phoenicis
3.2.1.163
additional information
Ca2+ is not required for activity
Aspergillus phoenicis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.2.1.114
74000
-
SDS-PAGE
Aspergillus phoenicis
3.2.1.114
81000
-
native-PAGE
Aspergillus phoenicis
3.2.1.114
97000
-
SDS-PAGE
Aspergillus phoenicis
3.2.1.114
110000
-
native-PAGE
Aspergillus phoenicis
3.2.1.163
74000
-
1 * 74000, SDS-PAGE
Aspergillus phoenicis
3.2.1.163
81000
-
gel filtration
Aspergillus phoenicis
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.2.1.114
anion-exchange chromatography, chromatofocusing chromatography, gel filtration
Aspergillus phoenicis
3.2.1.163
native enzyme 370fold by anion-exchange chromatography, chromatofocussing, and gel filtration
Aspergillus phoenicis
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.2.1.163
48
-
purified enzyme
Aspergillus phoenicis
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
3.2.1.163
23°C, room temperature, purified enzyme, over 95% remaining activity after 7 days
Aspergillus phoenicis
3.2.1.163
4°C, purified enzyme, over 95% remaining activity after 3 months
Aspergillus phoenicis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
3.2.1.114
alpha-D-Man-(1,6)-D-Man + H2O
-
664527
Aspergillus phoenicis
alpha-D-mannopyranose
-
-
-
?
3.2.1.114
alpha-D-Manp-(1,3)-D-Manp + H2O
-
664527
Aspergillus phoenicis
alpha-D-mannopyranose
-
-
-
?
3.2.1.114
alpha-D-Manp-(1,6)-D-Manp + H2O
-
664527
Aspergillus phoenicis
alpha-D-mannopyranose
-
-
-
?
3.2.1.114
baker's yeast mannan + H2O
-
664527
Aspergillus phoenicis
?
-
-
-
?
3.2.1.114
p-nitrophenyl-alpha-D-mannopyranoside + H2O
-
664527
Aspergillus phoenicis
p-nitrophenol + alpha-D-mannopyranose
-
-
-
?
3.2.1.163
alpha-D-Manp-(1,6)-D-Manp + H2O
the enzyme cleaves specifically (1-6)-linked mannobiose and has no activity towards any other linkages
664527
Aspergillus phoenicis
alpha-D-mannose
-
-
-
?
3.2.1.163
additional information
no activity with p-nitrophenyl-alpha-D-mannopyranoside or baker's yeast mannan
664527
Aspergillus phoenicis
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.2.1.163
monomer
1 * 74000, SDS-PAGE
Aspergillus phoenicis
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.2.1.114
60
-
-
Aspergillus phoenicis
3.2.1.163
60
-
-
Aspergillus phoenicis
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.2.1.163
30
-
purified enzyme, stable for 24 h, rapid inactivation above
Aspergillus phoenicis
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.2.1.163
4
4.5
-
Aspergillus phoenicis
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.2.1.114
4
4.5
-
Aspergillus phoenicis
3.2.1.163
3
5
36% of maximal activity at pH 3.0, rapid inactivation above pH 5.0
Aspergillus phoenicis
pI Value (protein specific)
EC Number
Organism
Commentary
pI Value Maximum
pI Value
3.2.1.114
Aspergillus phoenicis
for the first isoform of the enzyme, isoelectric focusing
-
4.3
3.2.1.114
Aspergillus phoenicis
method not mentioned
-
4.6
3.2.1.114
Aspergillus phoenicis
for the second isoform of the enzyme, isoelectric focusing
-
4.8
3.2.1.163
Aspergillus phoenicis
isoelectric focusing
-
4.6