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Literature summary for 3.2.1.20 extracted from

  • Chang, S.T.; Parker, K.N.; Bauer, M.W.; Kelly, R.M.
    alpha-Glucosidase from Pyrococcus furiosus (2001), Methods Enzymol., 330, 260-269.
    View publication on PubMed

Application

Application Comment Organism
nutrition usage in industrial processes to hydrolyze starch, saccharification step Pyrococcus furiosus

Cloned(Commentary)

Cloned (Comment) Organism
functional overexpression in Escherichia coli Pyrococcus furiosus

General Stability

General Stability Organism
DTT and urea stabilize against thermal inactivation Pyrococcus furiosus
guanidine hydrochloride inactivates Pyrococcus furiosus

Inhibitors

Inhibitors Comment Organism Structure
EDTA
-
Pyrococcus furiosus
guanidine hydrochloride inactivation Pyrococcus furiosus
SDS partial inactivation Pyrococcus furiosus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information thermal dependence of kinetics, overview Pyrococcus furiosus

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular
-
Thermococcus sp.
-
-
intracellular
-
Saccharolobus solfataricus 5622
-
intracellular
-
Pyrococcus woesei 5622
-

Metals/Ions

Metals/Ions Comment Organism Structure
additional information no requirement for Ca2+ Pyrococcus furiosus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
60000
-
1 * 60000 Thermococcus sp.
80000
-
x * 80000 Saccharolobus solfataricus
125000
-
-
Pyrococcus furiosus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
maltose + H2O Pyrococcus furiosus
-
2 D-glucose
-
?
additional information Pyrococcus furiosus the enzyme catalyzes the final step in the metabolism of carbohydrates, such as starch, to glucose for cell utilization, the oligosaccharide transport system shows high affinity for trehalose and maltose ?
-
?

Organism

Organism UniProt Comment Textmining
Pyrococcus furiosus
-
strain DSM 3638
-
Pyrococcus woesei
-
-
-
Saccharolobus solfataricus
-
-
-
Saccharolobus solfataricus 98/2
-
-
-
Thermococcus sp.
-
-
-
Thermococcus sp. AN1
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Thermococcus sp.
2 different protocols, 5-step procedure including ammonium sulfate precipitation and several chromatographic steps, 310fold Pyrococcus furiosus

Source Tissue

Source Tissue Comment Organism Textmining
additional information continuous culture, growth on yeast extract, tryptone and starch Pyrococcus furiosus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
185
-
purified enzyme, substrate methyl alpha-D-glucopyranoside Pyrococcus furiosus
287
-
purified enzyme, substrate 4-nitrophenyl alpha-D-glucopyranoside Pyrococcus furiosus
951
-
purified enzyme, substrate isomaltose Pyrococcus furiosus
1760
-
purified enzyme, substrate maltose Pyrococcus furiosus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl alpha-D-glucopyranoside + H2O
-
Pyrococcus furiosus 4-nitrophenol + alpha-D-glucopyranose
-
?
4-nitrophenyl alpha-D-glucopyranoside + H2O preferred substrate Thermococcus sp. 4-nitrophenol + alpha-D-glucopyranose
-
?
4-nitrophenyl alpha-D-glucopyranoside + H2O preferred substrate Thermococcus sp. AN1 4-nitrophenol + alpha-D-glucopyranose
-
?
isomaltose + H2O
-
Pyrococcus furiosus 2 D-glucose
-
?
maltopentaose + H2O preferred substrate Pyrococcus woesei D-glucose + ?
-
?
maltose + H2O
-
Pyrococcus furiosus 2 D-glucose
-
?
maltose + H2O best substrate Pyrococcus furiosus 2 D-glucose
-
?
maltose + H2O preferred substrate Saccharolobus solfataricus 2 D-glucose
-
?
maltose + H2O preferred substrate Saccharolobus solfataricus 98/2 2 D-glucose
-
?
methyl alpha-D-glucopyranoside + H2O
-
Pyrococcus furiosus methanol + alpha-D-glucopyranose
-
?
additional information the enzyme catalyzes the final step in the metabolism of carbohydrates, such as starch, to glucose for cell utilization, the oligosaccharide transport system shows high affinity for trehalose and maltose Pyrococcus furiosus ?
-
?
additional information sucrose is a poor or no substrate, no activity with starch Pyrococcus furiosus ?
-
?
oligosaccharides + H2O various substrates, enzyme is exo-acting to release alpha-D-glucose, preferential cleavage of the alpha-1,4-bond Pyrococcus furiosus alpha-D-glucose
-
?

Subunits

Subunits Comment Organism
dimer 2 * 55000-60000, SDS-PAGE Pyrococcus furiosus
monomer 1 * 60000 Thermococcus sp.
tetramer or pentamer x * 80000 Saccharolobus solfataricus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
98
-
at pH 5.0-6.0 Pyrococcus furiosus
105
-
-
Saccharolobus solfataricus
105
-
-
Pyrococcus woesei
105 115 at pH 5.5 Pyrococcus furiosus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
95
-
purified enzyme, stable 3 h Thermococcus sp.
98
-
half-life 30 min Thermococcus sp.
98
-
half-life 160-180 min Saccharolobus solfataricus
98
-
pH 6.5, loss of 50% activity within 48 h, in presence of 0.1 M DTT or 1 M urea 80% activity are retained for 30 min, enzyme shows loss of 78% activity in presence of 1 M SDS within 30 min, and loss of 50% activity in presence of 20 mM EDTA, complete inactivation occurs in presence of 1 M guanidine hydrochloride within 30 min Pyrococcus furiosus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4.5
-
-
Saccharolobus solfataricus
5 5.5
-
Pyrococcus woesei
5.5
-
assay at Pyrococcus furiosus
7.5
-
-
Thermococcus sp.