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Literature summary for 3.2.1.51 extracted from
- The alpha-L: -fucosidase from Sulfolobus solfataricus (2008), Extremophiles, 12, 61-68.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli | Saccharolobus solfataricus |
| expression in Escherichia coli | Saccharolobus solfataricus |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
- |
Thermotoga maritima |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D124G | activity is not significantly affected | Saccharolobus solfataricus |
| D124G | kcat/KM for 4-nitrophenyl-alpha-L-fucopyranoside is 3.7fold lower than wild-type value | Saccharolobus solfataricus |
| D146G | activity is not significantly affected | Saccharolobus solfataricus |
| D146G | kcat/KM for 4-nitrophenyl-alpha-L-fucopyranoside is 1.5fold lower than wild-type value | Saccharolobus solfataricus |
| D242G | inactive, reactivating in the presence of sodium azide or sodium formate, reactivated enzyme maintains its thermophilicity | Saccharolobus solfataricus |
| D242G | turnover number on 4-nitrophenyl-alpha-L-fucopyranoside is 0.0012times that of the wild type activity, 40fold reactivation by azide. The fucosyl-azide product obtained by the D242G mutant is in the inverted (beta-L-) configuration when compared with the substrate | Saccharolobus solfataricus |
| E292G | affects catalysis severely, unchanged affinity for 4-nitrophenyl-alpha-L-fucopyranoside but a 154fold reduction in the turnover number | Saccharolobus solfataricus |
| E292G | kcat/KM for 4-nitrophenyl-alpha-L-fucopyranoside is 311fold lower than wild-type value | Saccharolobus solfataricus |
| E58G | affects catalysis severely | Saccharolobus solfataricus |
| E58G | kcat/KM for 4-nitrophenyl-alpha-L-fucopyranoside is 5395fold lower than wild-type value | Saccharolobus solfataricus |
| H123G | affinity for the substrate is remarkably different from that of the wild-type | Saccharolobus solfataricus |
| H123G | kcat/KM for 4-nitrophenyl-alpha-L-fucopyranoside is 2929fold lower than wild-type value | Saccharolobus solfataricus |
| H46G | affinity for the substrate is remarkably different from that of the wild-type, 607fold increase in the KM | Saccharolobus solfataricus |
| H46G | kcat/KM for 4-nitrophenyl-alpha-L-fucopyranoside is 410fold lower than wild-type value | Saccharolobus solfataricus |
| additional information | mutations of the catalytic residues with non-nucleophilic amino acids lead to strong reduction or even abolition of the enzymatic activity. Mutants can be reactivated in the presence of external nucleophiles such as sodium azide | Saccharolobus solfataricus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.028 | - |
4-nitrophenyl-alpha-L-fucopyranoside | wild-type enzyme | Saccharolobus solfataricus |  |
| 0.028 | - |
4-nitrophenyl-alpha-L-fucopyranoside | wild-type, at pH 6.3, in the presence of sodium phosphate | Saccharolobus solfataricus | |
| 0.033 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant D146G | Saccharolobus solfataricus | |
| 0.033 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant enzyme D146G | Saccharolobus solfataricus | |
| 0.06 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant E292G, at pH 6.3, in the presence of sodium phosphate | Saccharolobus solfataricus | |
| 0.06 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant enzyme E292G | Saccharolobus solfataricus | |
| 0.09 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant D124G | Saccharolobus solfataricus | |
| 0.09 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant E292G, at pH 4.6, in the presence of sodium acetate | Saccharolobus solfataricus | |
| 0.09 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant enzyme D124G | Saccharolobus solfataricus | |
| 0.26 | - |
4-nitrophenyl-alpha-L-fucopyranoside | wild-type, at pH 5.0, in the presence of sodium citrate | Saccharolobus solfataricus | |
| 0.6 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant E58G, at pH 4.6, in the presence of sodium acetate | Saccharolobus solfataricus | |
| 1.1 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant E58G, at pH 4.6, in the presence of sodium formate | Saccharolobus solfataricus | |
| 1.6 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant E58G, at pH 4.6, in the presence of sodium citrate | Saccharolobus solfataricus | |
| 2.9 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant E58G, at pH 6.3, in the presence of sodium phosphate and NaN3 | Saccharolobus solfataricus | |
| 17 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant enzyme H46G | Saccharolobus solfataricus | |
| 17 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant H46G | Saccharolobus solfataricus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 57000 | - |
9 * 57000 | Saccharolobus solfataricus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Saccharolobus solfataricus | - |
- |
- |
| Thermotoga maritima | - |
- |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl-alpha-L-fucopyranoside + H2O | - |
Saccharolobus solfataricus | 4-nitrophenol + alpha-L-fucose | - |
? | |
| 4-nitrophenyl-alpha-L-fucopyranoside + N-acetyl-D-glucosamine + H2O | catalytic residues Asp242, Glu58, and Glu292 | Saccharolobus solfataricus | 4-nitrophenol + alpha-L-fucose-(1-3)-D-GlcNAc | - |
? | |
| alpha-L-Fuc-(1-3)-a-L-Fuc-O-4-NP + H2O | - |
Saccharolobus solfataricus | ? | - |
? | |
| alpha-L-Fuc-(1-3)-alpha-L-Fuc-O-4-nitrophenyl ester + H2O | - |
Saccharolobus solfataricus | ? | - |
? | |
| additional information | behaviour of the catalytic residues is different from that of Sulfolobus solfataricus alpha-fuc | Thermotoga maritima | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| nonamer | 9 * 57000 | Saccharolobus solfataricus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| alpha-fuc | - |
Thermotoga maritima |
| alpha-fuc | encoded by an interrupted gene | Saccharolobus solfataricus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 95 | - |
- |
Saccharolobus solfataricus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 80 | - |
2 h, 40% loss of activity | Saccharolobus solfataricus |
| 80 | - |
displays high stability maintaining 60% of the residual activity after 2 h | Saccharolobus solfataricus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.24 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant D242G | Saccharolobus solfataricus | |
| 0.24 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant enzyme D242G | Saccharolobus solfataricus | |
| 1.86 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant E292G at pH 6.3 in the presence of sodium phosphate or at pH 4.6 in the presence of sodium acetate | Saccharolobus solfataricus | |
| 1.86 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant enzyme E292G | Saccharolobus solfataricus | |
| 9.66 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant D242G, in the presence of 2 M sodium azide | Saccharolobus solfataricus | |
| 143 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant E58G, at pH 4.6, in the presence of sodium citrate | Saccharolobus solfataricus | |
| 224 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant D146G | Saccharolobus solfataricus | |
| 224 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant enzyme D146G | Saccharolobus solfataricus | |
| 240 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant D124G | Saccharolobus solfataricus | |
| 240 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant enzyme D124G | Saccharolobus solfataricus | |
| 287 | - |
4-nitrophenyl-alpha-L-fucopyranoside | wild-type enzyme | Saccharolobus solfataricus | |
| 287 | - |
4-nitrophenyl-alpha-L-fucopyranoside | wild-type, at pH 6.3, in the presence of sodium phosphate | Saccharolobus solfataricus | |
| 419 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant enzyme H46G | Saccharolobus solfataricus | |
| 419 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant H46G | Saccharolobus solfataricus | |
| 430 | - |
4-nitrophenyl-alpha-L-fucopyranoside | wild-type, at pH 5.0 | Saccharolobus solfataricus | |
| 586 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant E58G, at pH 4.6, in the presence of sodium acetate | Saccharolobus solfataricus | |
| 679 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant E58G, at pH 6.3, in the presence of sodium phosphate and NaN3 | Saccharolobus solfataricus | |
| 846 | - |
4-nitrophenyl-alpha-L-fucopyranoside | mutant E58G, at pH 4.6, in the presence of sodium formate | Saccharolobus solfataricus | |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 4.6 | 6.3 | - |
Saccharolobus solfataricus |
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