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Literature summary for 3.2.1.B26 extracted from

  • Moracci, M.; Capalbo, L.; Ciaramella, M.; Rossi, M.
    Identification of two glutamic acid residues essential for catalysis in the beta-glycosidase from the thermoacidophilic archaeon Sulfolobus solfataricus (1996), Protein Eng., 9, 1191-1195.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
E206Q mutant shows 10- and 60-fold reduced activities on aryl-galacto and aryl-glucosides, respectively, when compared with the wild type. Significant decrease in Km-value with 4-nitrophenyl beta-D-glucoside or 2-nitrophenyl beta-D-glucoside. The residual activity of the mutant loses the typical pH dependence shown by the wild type. These data suggest that Glu206 acts as the general acid/base catalyst in the hydrolysis reaction Saccharolobus solfataricus
E206Q mutant shows 10- and 60-fold reduced activities on aryl-galacto and aryl-glucosides, respectively, when compared with the wild type. Significant decrease in Km-value with 4-nitrophenyl beta-D-glucoside or 2-nitrophenyl betaD-glucoside. The residual activity of the mutant loses the typical pH dependence shown by the wild type. These data suggest that Glu206 acts as the general acid/base catalyst in the hydrolysis reaction Saccharolobus solfataricus
E387Q mutant shows no detectable activity Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.03
-
4-nitrophenyl beta-D-glucoside pH 6.5, 65°C, mutant enzyme E206Q Saccharolobus solfataricus
0.03
-
2-Nitrophenyl beta-D-glucoside pH 6.5, 65°C, mutant enzyme E206Q Saccharolobus solfataricus
0.09
-
4-nitrophenyl beta-D-fucoside pH 6.5, 65°C, mutant enzyme E206Q Saccharolobus solfataricus
0.3
-
4-nitrophenyl beta-D-glucoside pH 6.5, 65°C, wild-type enzyme Saccharolobus solfataricus
0.45
-
4-nitrophenyl beta-D-fucoside pH 6.5, 65°C, wild-type enzyme Saccharolobus solfataricus
0.95
-
2-nitrophenyl beta-D-galactopyranoside pH 6.5, 65°C, wild-type enzyme Saccharolobus solfataricus
1.01
-
2-Nitrophenyl beta-D-glucoside pH 6.5, 65°C, wild-type enzyme Saccharolobus solfataricus
1.17
-
4-nitrophenyl beta-D-galactopyranoside pH 6.5, 65°C, wild-type enzyme Saccharolobus solfataricus
1.53
-
2-nitrophenyl beta-D-galactopyranoside pH 6.5, 65°C, mutant enzyme E206Q Saccharolobus solfataricus
4.79
-
4-nitrophenyl beta-D-galactopyranoside pH 6.5, 65°C, mutant enzyme E206Q Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus
-
-
-
Saccharolobus solfataricus MT4
-
-
-

Purification (Commentary)

Purification (Comment) Organism
purification of mutant proteins E206Q and E387Q to homogeneity using the Schistosoma japonicum glutathione S-transferase fusion system Saccharolobus solfataricus
purification of mutant proteins E206Q and E387Q to homogeneity using the Schistosoma japonicum glutathione-S-transferase fusion system Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-nitrophenyl beta-D-galactopyranoside + H2O
-
Saccharolobus solfataricus 2-nitrophenol + beta-D-galactopyranose
-
?
2-nitrophenyl beta-D-galactopyranoside + H2O
-
Saccharolobus solfataricus MT4 2-nitrophenol + beta-D-galactopyranose
-
?
2-nitrophenyl beta-D-glucoside + H2O
-
Saccharolobus solfataricus 2-nitrophenol + beta-D-glucose
-
?
2-nitrophenyl beta-D-glucoside + H2O
-
Saccharolobus solfataricus MT4 2-nitrophenol + beta-D-glucose
-
?
4-nitrophenyl beta-D-fucoside + H2O
-
Saccharolobus solfataricus 4-nitrophenol + beta-D-fucose
-
?
4-nitrophenyl beta-D-fucoside + H2O
-
Saccharolobus solfataricus MT4 4-nitrophenol + beta-D-fucose
-
?
4-nitrophenyl beta-D-galactopyranoside + H2O
-
Saccharolobus solfataricus 4-nitrophenol + beta-D-galactopyranose
-
?
4-nitrophenyl beta-D-galactopyranoside + H2O
-
Saccharolobus solfataricus MT4 4-nitrophenol + beta-D-galactopyranose
-
?
4-nitrophenyl beta-D-glucoside + H2O
-
Saccharolobus solfataricus 4-nitrophenol + beta-D-glucose
-
?
4-nitrophenyl beta-D-glucoside + H2O
-
Saccharolobus solfataricus MT4 4-nitrophenol + beta-D-glucose
-
?

Synonyms

Synonyms Comment Organism
Sbetagly
-
Saccharolobus solfataricus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
65
-
assay at Saccharolobus solfataricus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.4
-
4-nitrophenyl beta-D-glucoside pH 6.5, 65°C, mutant enzyme E206Q Saccharolobus solfataricus
4.5
-
2-Nitrophenyl beta-D-glucoside pH 6.5, 65°C, mutant enzyme E206Q Saccharolobus solfataricus
31
-
4-nitrophenyl beta-D-galactopyranoside pH 6.5, 65°C, mutant enzyme E206Q Saccharolobus solfataricus
34
-
2-nitrophenyl beta-D-galactopyranoside pH 6.5, 65°C, mutant enzyme E206Q Saccharolobus solfataricus
35
-
4-nitrophenyl beta-D-fucoside pH 6.5, 65°C, mutant enzyme E206Q Saccharolobus solfataricus
240
-
4-nitrophenyl beta-D-glucoside pH 6.5, 65°C, wild-type enzyme Saccharolobus solfataricus
252
-
2-Nitrophenyl beta-D-glucoside pH 6.5, 65°C, wild-type enzyme Saccharolobus solfataricus
263
-
4-nitrophenyl beta-D-fucoside pH 6.5, 65°C, wild-type enzyme Saccharolobus solfataricus
275
-
4-nitrophenyl beta-D-galactopyranoside pH 6.5, 65°C, wild-type enzyme Saccharolobus solfataricus
295
-
2-nitrophenyl beta-D-galactopyranoside pH 6.5, 65°C, wild-type enzyme Saccharolobus solfataricus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
-
Saccharolobus solfataricus

pH Range

pH Minimum pH Maximum Comment Organism
5.7 8 pH 5.6: about 45% of maximal activity, pH 8.0: about 55% of maximal activity Saccharolobus solfataricus