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Literature summary for 3.2.1.101 extracted from

  • Nakajima, T.; Maitra, S.K.; Ballou, C.E.
    An endo-alpha1 -> 6-D-mannanase from a soil bacterium. Purification, properties, and mode of action (1976), J. Biol. Chem., 251, 174-181.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
ethanol 1% v/v ethanol: slight activation Niallia circulans
iodoacetamide 1 mM: slight activation Niallia circulans
N-ethylmaleimide 1 mM: slight activation Niallia circulans
Propanol slight activation Niallia circulans

Application

Application Comment Organism
analysis used to determine the structure of mannan-protein linkage region Niallia circulans

Inhibitors

Inhibitors Comment Organism Structure
Cu2+
-
Niallia circulans
diisopropyl fluorophosphate
-
Niallia circulans
EDTA 70% inhibition, 90% of original activity restored with 10 mM Ca2+ Niallia circulans
ethanol 10% ethanol: inactivation Niallia circulans
Fe2+
-
Niallia circulans
iodoacetamide 2 mM: 50% inhibition Niallia circulans
Mannan Saccharomyces cerevisiae wild type mannan, 20% inhibition Niallia circulans
Mg2+ very slight inhibition Niallia circulans
N-ethylmaleimide 2 mM: 50% inhibition Niallia circulans
p-chloromercuribenzoate 1 mM: complete inhibition, 0.001 mM: 50% inhibition Niallia circulans
reduced alpha-1,6-mannohexaose 54% inhibition, also serves as a substrate Niallia circulans
reduced alpha-1,6-mannotriose competitive inhibitor, 92% inhibition Niallia circulans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetic data Niallia circulans
0.0044
-
alpha-1,6-D-mannan
-
Niallia circulans
0.072
-
alpha-1,6-D-mannohexaose
-
Niallia circulans
0.12
-
alpha-1,6-D-mannopentaose
-
Niallia circulans
0.33
-
alpha-1,6-D-mannotetraose
-
Niallia circulans
1
-
alpha-1,6-D-mannotriose
-
Niallia circulans
4
-
alpha-1,6-D-mannobiose
-
Niallia circulans

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular
-
Niallia circulans
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ activation, required for full activity Niallia circulans
Co2+ very slight activation Niallia circulans
Mn2+ activation Niallia circulans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
125000
-
gel filtration and SDS-PAGE Niallia circulans
131000
-
sedimentation equilibrium Niallia circulans

Organic Solvent Stability

Organic Solvent Comment Organism
Ethanol 1% v/v ethanol: stable and slight activation, effect decreases at higher concentrations, at 10% ethanol: inactivation Niallia circulans
propanol stable and slight activation Niallia circulans

Organism

Organism UniProt Comment Textmining
Niallia circulans
-
Bacillus sp. TN-31: probably Bacillus circulans
-

Purification (Commentary)

Purification (Comment) Organism
-
Niallia circulans

Reaction

Reaction Comment Organism Reaction ID
(1,6-beta-D-mannopyranosyl)n-1,6-beta-D-mannopyranose + H2O = (1,6-beta-D-mannopyranosyl)n-2-1,6-beta-D-mannopyranose + 6-O-beta-D-mannopyranosyl-beta-D-mannopyranose enzyme hydrolyzes alpha-1,6-D-mannosidic and not beta-1,6-D-mannosidic linkages Niallia circulans

Source Tissue

Source Tissue Comment Organism Textmining
culture filtrate
-
Niallia circulans
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.87
-
-
Niallia circulans

Storage Stability

Storage Stability Organism
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 Niallia circulans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
alpha-1,6-D-mannan + H2O unbranched, with more than 80% alpha-1,6-D-mannosidic linkages, best substrate Niallia circulans D-mannose + alpha-1,6-D-mannobiose end-product, with intermediate formation of alpha-1,6-mannooligosaccharides of various sizes ?
alpha-1,6-D-mannan + H2O Saccharomyces cerevisiae X2180-1A backbone mannan Niallia circulans D-mannose + alpha-1,6-D-mannobiose end-product, with intermediate formation of alpha-1,6-mannooligosaccharides of various sizes ?
alpha-1,6-D-mannan + H2O Saccharomyces cerevisiae X2180-1A-5 mannan Niallia circulans D-mannose + alpha-1,6-D-mannobiose end-product, with intermediate formation of alpha-1,6-mannooligosaccharides of various sizes ?
alpha-1,6-D-mannohexaose + H2O
-
Niallia circulans D-mannose + alpha-1,6-D-mannobiose 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 largest substrate Niallia circulans D-mannose + alpha-1,6-D-mannobiose 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 smallest substrate: alpha-1,6-mannotriose, largest substrate: alpha-1,6-mannooctaose Niallia circulans mannose + alpha-1,6-D-mannobiose end-product, with intermediate formation of mannose, mannobiose and mannotriose, the latter apparently converts more slowly to the smaller products ?
alpha-1,6-D-mannopentaose + H2O
-
Niallia circulans D-mannose + alpha-1,6-D-mannobiose 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
-
Niallia circulans D-mannose + alpha-1,6-D-mannobiose 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 smallest substrate Niallia circulans D-mannose + alpha-1,6-D-mannobiose
-
?
additional information substrate specificity Niallia circulans ?
-
?
additional information enzyme hydrolyzes alpha-1,6-D-mannosidic and not beta-1,6-D-mannosidic linkages Niallia circulans ?
-
?
additional information no activity on branched alpha-mannan, alpha-1,6-mannans that contain side chains, little or no activity with glucans: amylose, dextran, cellulose, laminarin, alpha-1,6-mannobiose, reduced mannobiose, reduced mannotriose, no activity with methyl alpha-D-mannoside or p-nitrophenyl alpha-D-mannoside, no activity with beta-1,4-D-mannan, methyl alpha-D-mannopyranoside or p-nitrophenyl alpha-D-mannopyranoside Niallia circulans ?
-
?
reduced mannohexaose + H2O poor substrate Niallia circulans ?
-
?
reduced mannooctaose + H2O poor substrate Niallia circulans ?
-
?
reduced mannopentaose + H2O poor substrate Niallia circulans ?
-
?
reduced mannotetraose + H2O poor substrate Niallia circulans ?
-
?

Subunits

Subunits Comment Organism
monomer 1 * 125000, SDS-PAGE Niallia circulans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
pH 5.5 Niallia circulans

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
unusually heat-stable Niallia circulans
55
-
stable at, in the presence of substrate, for at least 3 h Niallia circulans
60 65 5 min, denatured Niallia circulans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
7.33
-
alpha-1,6-D-mannan
-
Niallia circulans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5 7
-
Niallia circulans

pH Range

pH Minimum pH Maximum Comment Organism
5 7.5
-
Niallia circulans

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.001
-
p-chloromercuribenzoate
-
Niallia circulans
2
-
N-ethylmaleimide
-
Niallia circulans
2
-
iodoacetamide
-
Niallia circulans