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Literature summary extracted from

  • Pouwels, J.; Moracci, M.; Cobucci-Ponzano, B.; Perugino, G.; van der Oost, J.; Kaper, T.; Lebbink, J.H.; de Vos, W.M.; Ciaramella, M.; Rossi, M.
    Activity and stability of hyperthermophilic enzymes: a comparative study on two archaeal beta-glycosidases (2000), Extremophiles, 4, 157-164.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.2.1.B26 SDS 50°C, 30 min, 0.03%, slight activation Saccharolobus solfataricus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.B28 expression in Escherichia coli Pyrococcus furiosus

General Stability

EC Number General Stability Organism
3.2.1.B26 chaotropes (I-, ClO4-, NO3- and Cs+) have a strong destabilising effect on the protein by perturbing hydrophobic interactions Saccharolobus solfataricus
3.2.1.B26 denaturing effect of NaCl and Na2SO4 Saccharolobus solfataricus
3.2.1.B28 chaotropes (I-, ClO4-, NO3- and Cs+) have a strong destabilising effect on the protein by perturbing hydrophobic interactions Pyrococcus furiosus
3.2.1.B28 denaturing effect of NaCl Pyrococcus furiosus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.B26 SDS 50°C, 30 min, 0.3% SDS, 40% loss of activity Saccharolobus solfataricus
3.2.1.B28 SDS 0.03%, 50% residual activity. 0.04% less than 10% residual activity Pyrococcus furiosus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.B26 additional information
-
 additional information KM increases with temperature: the increase is moderate at lower temperatures (from 30°C to 65°C) and more evident between 65°C and 85°C. Reactions with 4-nitrophenyl-beta-D-glucoside and 2-nitrophenyl-beta-D-glucoside show a biphasic behavior in Lineweaver–Burk plots, suggesting that transglycosylation reactions occur at higher substrate concentrations. Kinetic constants obtained at lower or higher substrate concentrations are calculated Saccharolobus solfataricus
3.2.1.B26 0.59
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 30°C Saccharolobus solfataricus
3.2.1.B26 0.72
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 60°C Saccharolobus solfataricus
3.2.1.B26 0.77
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 65°C Saccharolobus solfataricus
3.2.1.B26 0.82
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 40°C Saccharolobus solfataricus
3.2.1.B26 0.87
-
 2-nitrophenyl beta-D-galactoside pH 5.4, 75°C Saccharolobus solfataricus
3.2.1.B26 1.09
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 50°C Saccharolobus solfataricus
3.2.1.B26 1.61
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 75°C Saccharolobus solfataricus
3.2.1.B26 3.16
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 85°C Saccharolobus solfataricus
3.2.1.B28 0.25
-
 2-nitrophenyl beta-D-glucopyranoside pH 5.4, 75°C Pyrococcus furiosus
3.2.1.B28 0.37
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.4, 75°C Pyrococcus furiosus
3.2.1.B28 1.11
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 30°C Pyrococcus furiosus
3.2.1.B28 1.31
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 40°C Pyrococcus furiosus
3.2.1.B28 1.97
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 65°C Pyrococcus furiosus
3.2.1.B28 2.05
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 50°C Pyrococcus furiosus
3.2.1.B28 2.09
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 60°C Pyrococcus furiosus
3.2.1.B28 2.57
-
 2-nitrophenyl beta-D-galactopyranoside pH 5.4, 75°C Pyrococcus furiosus
3.2.1.B28 2.8
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 75°C Pyrococcus furiosus
3.2.1.B28 4.1
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 85°C Pyrococcus furiosus

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.B26 Saccharolobus solfataricus
-
-
-
3.2.1.B28 Pyrococcus furiosus Q51723
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.B26
-
 Saccharolobus solfataricus
3.2.1.B28
-
 Pyrococcus furiosus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.B26 2-nitrophenyl beta-D-galactoside + H2O
-
 Saccharolobus solfataricus 2-nitrophenol + beta-D-galactose
-
 ?
3.2.1.B26 4-nitrophenyl beta-D-galactoside + H2O
-
 Saccharolobus solfataricus 4-nitrophenol + beta-D-galactose
-
 ?
3.2.1.B26 4-nitrophenyl beta-D-glucoside + H2O
-
 Saccharolobus solfataricus 4-nitrophenol + beta-D-glucose
-
 ?
3.2.1.B28 2-nitrophenyl beta-D-galactopyranoside + H2O
-
 Pyrococcus furiosus 2-nitrophenol + beta-D-galactopyranose
-
 ?
3.2.1.B28 2-nitrophenyl beta-D-glucopyranoside + H2O
-
 Pyrococcus furiosus 2-nitrophenol + D-glucopyranose
-
 ?
3.2.1.B28 4-nitrophenyl beta-D-galactopyranoside + H2O
-
 Pyrococcus furiosus 4-nitrophenol + beta-D-galactopyranose
-
 ?
3.2.1.B28 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Pyrococcus furiosus 4-nitrophenol + D-glucopyranose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.B26 Sbetagly
-
 Saccharolobus solfataricus
3.2.1.B28 CelB
-
 Pyrococcus furiosus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.B26 additional information
-
 inactivation is coupled to irreversible aggregation Saccharolobus solfataricus
3.2.1.B26 75
-
 30 min, 90% residual activity Saccharolobus solfataricus
3.2.1.B28 75
-
 30 min, completely stable Pyrococcus furiosus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.B26 additional information
-
 additional information kcat increases exponentially with temperature. Reactions with 4-nitrophenyl-beta-D-glucoside and 2-nitrophenyl-beta-D-glucoside show a biphasic behavior in Lineweaver–Burk plots, suggesting that transglycosylation reactions occur at higher substrate concentrations. Kinetic constants obtained at lower or higher substrate concentrations are calculated Saccharolobus solfataricus
3.2.1.B26 56
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 30°C Saccharolobus solfataricus
3.2.1.B26 143
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 40°C Saccharolobus solfataricus
3.2.1.B26 293
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 50°C Saccharolobus solfataricus
3.2.1.B26 648
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 60°C Saccharolobus solfataricus
3.2.1.B26 1000
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 65°C Saccharolobus solfataricus
3.2.1.B26 2288
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 75°C Saccharolobus solfataricus
3.2.1.B26 2400
-
 2-nitrophenyl beta-D-galactoside pH 5.4, 75°C Saccharolobus solfataricus
3.2.1.B26 4380
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 85°C Saccharolobus solfataricus
3.2.1.B28 90
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 30°C Pyrococcus furiosus
3.2.1.B28 236
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 40°C Pyrococcus furiosus
3.2.1.B28 580
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 50°C Pyrococcus furiosus
3.2.1.B28 1222
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 60°C Pyrococcus furiosus
3.2.1.B28 1486
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 65°C Pyrococcus furiosus
3.2.1.B28 2600
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.4, 75°C Pyrococcus furiosus
3.2.1.B28 2907
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 75°C Pyrococcus furiosus
3.2.1.B28 3300
-
 2-nitrophenyl beta-D-glucopyranoside pH 5.4, 75°C Pyrococcus furiosus
3.2.1.B28 4863
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 85°C Pyrococcus furiosus
3.2.1.B28 9600
-
 2-nitrophenyl beta-D-galactopyranoside pH 5.4, 75°C Pyrococcus furiosus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.B26 4.5 6.5 at 65°C Saccharolobus solfataricus
3.2.1.B26 5 5.5 at 75°C Saccharolobus solfataricus
3.2.1.B28 5.4
-
-
 Pyrococcus furiosus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.B26 additional information
-
 activity dependence on pH at 65°C and 75°C on 5 mM 4-nitrophenyl beta-D-galactoside Saccharolobus solfataricus
3.2.1.B26 4 7 pH 4.0: about 60% of maximal activity at 75°C, pH 7.0: about 50% of maximal activity at 75°C Saccharolobus solfataricus

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.B26 3.4
-
 75°C, half-life is less than 5 min, irreversible aggregation Saccharolobus solfataricus
3.2.1.B28 10
-
 75°C, 90 min, stable Pyrococcus furiosus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.21 1000
-
 4-nitrophenyl beta-D-galactopyranoside pH 5.4, 75°C Pyrococcus furiosus
3.2.1.21 3700
-
 2-nitrophenyl beta-D-galactopyranoside pH 5.4, 75°C Pyrococcus furiosus
3.2.1.21 7100
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.4, 75°C Pyrococcus furiosus
3.2.1.21 12900
-
 2-nitrophenyl beta-D-glucopyranoside pH 5.4, 75°C Pyrococcus furiosus
3.2.1.B26 additional information
-
 additional information the efficiency (kcat/KM) increases with temperature and reaches its maximal efficiency at 65°C. Reactions with 4-nitrophenyl-beta-D-glucoside and 2-nitrophenyl-beta-D-glucoside show a biphasic behavior in Lineweaver–Burk plots, suggesting that transglycosylation reactions occur at higher substrate concentrations. Kinetic constants obtained at lower or higher substrate concentrations are calculated Saccharolobus solfataricus
3.2.1.B26 1400
-
 4-nitrophenyl beta-D-galactoside pH 5.4, 75°C Saccharolobus solfataricus
3.2.1.B26 2700
-
 2-nitrophenyl beta-D-galactoside pH 5.4, 75°C Saccharolobus solfataricus