Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Saccharolobus solfataricus |
expression in Escherichia coli | Saccharolobus solfataricus |
Crystallization (Comment) | Organism |
---|---|
- |
Thermotoga maritima |
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 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 Maximum [Da] | Comment | Organism |
---|---|---|---|
57000 | - |
9 * 57000 | Saccharolobus solfataricus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharolobus solfataricus | - |
- |
- |
Thermotoga maritima | - |
- |
- |
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 | Comment | Organism |
---|---|---|
nonamer | 9 * 57000 | Saccharolobus solfataricus |
Synonyms | Comment | Organism |
---|---|---|
alpha-fuc | - |
Thermotoga maritima |
alpha-fuc | encoded by an interrupted gene | Saccharolobus solfataricus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
95 | - |
- |
Saccharolobus solfataricus |
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 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 Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4.6 | 6.3 | - |
Saccharolobus solfataricus |