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Literature summary for 3.2.1.B26 extracted from

  • Corbett, K.; Fordham-Skelton, A.P.; Gatehouse, J.A.; Davis, B.G.
    Tailoring the substrate specificity of the beta-glycosidase from the thermophilic archaeon Sulfolobus solfataricus (2001), FEBS Lett., 509, 355-360.
    View publication on PubMed
Search for more literature for 3.2.1.B26

Cloned(Commentary)

Cloned (Comment) Organism
wild-type, mutant E432C, mutant W433C and mutant M439C enzymes are expressed in Escherichia coli as recombinant proteins containing an N-terminal His6-tag Saccharolobus solfataricus

Protein Variants

Protein Variants Comment Organism
E432C kcat/KM is reduced for 4-methylumbelliferyl beta-D-glucopyranoside is reduced 200fold, kcat/KM is reduced for 4-methylumbelliferyl beta-D-galactopyranoside is reduced 130fold, kcat/KM is reduced for 4-methylumbelliferyl beta-D-mannopyranoside is reduced 16fold, kcat/KM is reduced for 4-methylumbelliferyl beta-D-xylopyranoside is reduced 14fold Saccharolobus solfataricus
M439C shows almost identical values to wild-type for 4-methylumbelliferyl beta-D-glucopyranoside, 4-methylumbelliferyl beta-D-galactopyranoside or 4-methylumbelliferyl beta-D-mannopyranoside substrates. kcat/KM for 4-methylumbelliferyl beta-D-fucopyranoside is 1.8-fold lower than wild-type value and kcat/KM for 4-methylumbelliferyl beta-D-xylopyranoside is 4.7fold higher than wild-type Saccharolobus solfataricus
W433C kcat/KM is reduced for 4-methylumbelliferyl beta-D-glucopyranoside is reduced 140fold, kcat/KM is reduced for 4-methylumbelliferyl beta-D-galactopyranoside is reduced 230fold, kcat/KM is reduced for 4-methylumbelliferyl beta-D-mannopyranoside is reduced 10fold, kcat/KM is reduced for 4-methylumbelliferyl beta-D-xylopyranoside is reduced 12fold Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.011
-
 4-methylumbelliferyl beta-D-fucopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
0.023
-
 4-methylumbelliferyl beta-D-fucopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
0.036
-
 4-methylumbelliferyl beta-D-mannopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
0.042
-
 4-methylumbelliferyl beta-D-mannopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
0.046
-
 4-methylumbelliferyl beta-D-glucopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
0.066
-
 4-methylumbelliferyl beta-D-galactopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
0.068
-
 4-methylumbelliferyl beta-D-glucopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
0.068
-
 4-methylumbelliferyl beta-D-xylopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
0.083
-
 4-methylumbelliferyl beta-D-galactopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
0.13
-
 4-methylumbelliferyl beta-D-xylopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
0.18
-
 4-methylumbelliferyl beta-D-mannopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
0.34
-
 4-methylumbelliferyl beta-D-glucopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
0.34
-
 4-methylumbelliferyl beta-D-fucopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
0.41
-
 4-methylumbelliferyl beta-D-fucopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
0.47
-
 4-methylumbelliferyl beta-D-galactopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
0.59
-
 4-methylumbelliferyl beta-D-xylopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
0.9
-
 4-methylumbelliferyl beta-D-mannopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
1.26
-
 4-methylumbelliferyl beta-D-xylopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
1.3
-
 4-methylumbelliferyl beta-D-glucuronic acid pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
1.4
-
 4-methylumbelliferyl beta-D-glucuronic acid pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
1.61
-
 4-methylumbelliferyl beta-D-glucopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
2.2
-
 4-methylumbelliferyl beta-D-galactopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
57000
-
 2 * 57000, SDS-PAGE Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
His6-tagged mutant enzyme Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-methylumbelliferyl beta-D-fucopyranoside + H2O
-
 Saccharolobus solfataricus 4-methylumbelliferone + beta-D-fucopyranose
-
 ?
4-methylumbelliferyl beta-D-galactopyranoside + H2O
-
 Saccharolobus solfataricus 4-methylumbelliferone + beta-D-galactopyranose
-
 ?
4-methylumbelliferyl beta-D-glucopyranoside + H2O
-
 Saccharolobus solfataricus 4-methylumbelliferone + beta-D-glucopyranose
-
 ?
4-methylumbelliferyl beta-D-glucuronic acid + H2O
-
 Saccharolobus solfataricus 4-methylumbelliferone + beta-D-glucuronic acid
-
 ?
4-methylumbelliferyl beta-D-mannopyranoside + H2O
-
 Saccharolobus solfataricus 4-methylumbelliferone + beta-D-mannopyranose
-
 ?
4-methylumbelliferyl beta-D-xylopyranoside + H2O
-
 Saccharolobus solfataricus 4-methylumbelliferone + beta-D-xylopyranose
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 57000, SDS-PAGE Saccharolobus solfataricus

Synonyms

Synonyms Comment Organism
LACS
-
 Saccharolobus solfataricus
SsbetaG
-
 Saccharolobus solfataricus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
 assay at Saccharolobus solfataricus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.81
-
 4-methylumbelliferyl beta-D-glucuronic acid pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
0.92
-
 4-methylumbelliferyl beta-D-mannopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
1.3
-
 4-methylumbelliferyl beta-D-glucuronic acid pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
1.5
-
 4-methylumbelliferyl beta-D-xylopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
1.8
-
 4-methylumbelliferyl beta-D-mannopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
2.3
-
 4-methylumbelliferyl beta-D-mannopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
2.8
-
 4-methylumbelliferyl beta-D-mannopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
2.8
-
 4-methylumbelliferyl beta-D-xylopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
3.8
-
 4-methylumbelliferyl beta-D-xylopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
5.1
-
 4-methylumbelliferyl beta-D-glucopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
5.4
-
 4-methylumbelliferyl beta-D-galactopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
9.3
-
 4-methylumbelliferyl beta-D-xylopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
14
-
 4-methylumbelliferyl beta-D-galactopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
18
-
 4-methylumbelliferyl beta-D-fucopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
31
-
 4-methylumbelliferyl beta-D-fucopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
33
-
 4-methylumbelliferyl beta-D-glucopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
80
-
 4-methylumbelliferyl beta-D-fucopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
91
-
 4-methylumbelliferyl beta-D-fucopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
94
-
 4-methylumbelliferyl beta-D-galactopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
98
-
 4-methylumbelliferyl beta-D-galactopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
140
-
 4-methylumbelliferyl beta-D-glucopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
190
-
 4-methylumbelliferyl beta-D-glucopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus

pH Optimum

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

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.6
-
 4-methylumbelliferyl beta-D-glucuronic acid pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
0.92
-
 4-methylumbelliferyl beta-D-glucuronic acid pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
2.2
-
 4-methylumbelliferyl beta-D-xylopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
2.5
-
 4-methylumbelliferyl beta-D-xylopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
3.2
-
 4-methylumbelliferyl beta-D-mannopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
5.1
-
 4-methylumbelliferyl beta-D-mannopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
6.3
-
 4-methylumbelliferyl beta-D-galactopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
11
-
 4-methylumbelliferyl beta-D-galactopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
15
-
 4-methylumbelliferyl beta-D-glucopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
20
-
 4-methylumbelliferyl beta-D-glucopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
30
-
 4-methylumbelliferyl beta-D-xylopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
50
-
 4-methylumbelliferyl beta-D-mannopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
53
-
 4-methylumbelliferyl beta-D-fucopyranoside pH 6.5, 80°C, mutant enzyme E432C Saccharolobus solfataricus
53
-
 4-methylumbelliferyl beta-D-mannopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
76
-
 4-methylumbelliferyl beta-D-fucopyranoside pH 6.5, 80°C, mutant enzyme W433C Saccharolobus solfataricus
136
-
 4-methylumbelliferyl beta-D-xylopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
1130
-
 4-methylumbelliferyl beta-D-galactopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
1490
-
 4-methylumbelliferyl beta-D-galactopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
2900
-
 4-methylumbelliferyl beta-D-glucopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
2900
-
 4-methylumbelliferyl beta-D-glucopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus
4000
-
 4-methylumbelliferyl beta-D-fucopyranoside pH 6.5, 80°C, mutant enzyme M439C Saccharolobus solfataricus
7300
-
 4-methylumbelliferyl beta-D-fucopyranoside pH 6.5, 80°C, wild-type enzyme Saccharolobus solfataricus