Protein Variants | Comment | Organism |
---|---|---|
N62Y/Q63K/L231F/V303A/A412V | mutations observed in all the clones isolated | Thermosphaera aggregans |
N62Y/Q63K/L231F/V303A/E386G/A412V | additional mutation in nucleophile residue E386, mutation completely abolishes activity. The addition of 2 M sodium formate as an external nucleophile leads to the recovery of 4.60% activity with accumulation of oligosaccharides. At pH 3.0 and low concentrations of sodium formate buffer, the hyperthermophilic glycosynthase shows kcat values similar to those of the wild-type and 17fold higher than those observed at the usual reactivation conditions in 2 M sodium formate at pH 6.5 | Thermosphaera aggregans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.2 | - |
2-nitrophenyl beta-D-glucopyranoside | wild-type, pH 6.5, 65°C | Thermosphaera aggregans | |
0.38 | - |
2-nitrophenyl beta-D-glucopyranoside | wild-type, pH 6.5, 65°C, presence of 2 M formate | Thermosphaera aggregans | |
1.01 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant E387G, pH 6.5, 65°C, presence of 2 M formate | Thermosphaera aggregans | |
2.8 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 6.0, 65°C | Thermosphaera aggregans | |
3.1 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 5.0, 65°C | Thermosphaera aggregans | |
3.3 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 4.0, 65°C | Thermosphaera aggregans | |
6.6 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 3.0, 65°C | Thermosphaera aggregans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermosphaera aggregans | Q9YGA8 | - |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.39 | - |
mutant N62Y/Q63K/L231F/V303A/E386G/A412V, substrate cellobiose, pH 3.0, 65°C | Thermosphaera aggregans |
6.3 | - |
mutant N62Y/Q63K/L231F/V303A/E386G/A412V, substrate 2-nitrophenyl beta-D-galactopyranoside, pH 3.0, 65°C | Thermosphaera aggregans |
6.4 | - |
mutant N62Y/Q63K/L231F/V303A/E386G/A412V, substrate 2-nitrophenyl beta-D-xylopyranoside, pH 3.0, 65°C | Thermosphaera aggregans |
10.4 | - |
wild-type, substrate 2-nitrophenyl beta-D-xylopyranoside, pH 3.0, 65°C | Thermosphaera aggregans |
12 | - |
mutant N62Y/Q63K/L231F/V303A/E386G/A412V, substrate 4-nitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C | Thermosphaera aggregans |
33.8 | - |
wild-type, substrate cellobiose, pH 3.0, 65°C | Thermosphaera aggregans |
61.7 | - |
wild-type, substrate 4-nitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C | Thermosphaera aggregans |
72.2 | - |
wild-type, substrate 2-nitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C | Thermosphaera aggregans |
75 | - |
wild-type, substrate 2,4-dinitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C | Thermosphaera aggregans |
129.8 | - |
mutant N62Y/Q63K/L231F/V303A/E386G/A412V, substrate 2-nitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C | Thermosphaera aggregans |
140.9 | - |
wild-type, substrate 2-nitrophenyl beta-D-galactopyranoside, pH 3.0, 65°C | Thermosphaera aggregans |
207.1 | - |
mutant N62Y/Q63K/L231F/V303A/E386G/A412V, substrate 2,4-dinitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C | Thermosphaera aggregans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,4-dinitrophenyl beta-D-glucopyranoside + H2O | - |
Thermosphaera aggregans | 2,4-dinitrophenol + D-glucopyranose | - |
? | |
2-nitrophenyl beta-D-galactopyranoside + H2O | - |
Thermosphaera aggregans | 2-nitrophenol + D-galactopyranose | - |
? | |
2-nitrophenyl beta-D-glucopyranoside + H2O | - |
Thermosphaera aggregans | 2-nitrophenol + D-glucopyranose | - |
? | |
2-nitrophenyl beta-D-xylopyranoside + H2O | - |
Thermosphaera aggregans | 2-nitrophenol + D-xylopyranose | - |
? | |
4-nitrophenyl beta-D-glucopyranoside + H2O | - |
Thermosphaera aggregans | 4-nitrophenol + D-glucopyranose | - |
? | |
cellobiose + H2O | - |
Thermosphaera aggregans | 2 D-glucose | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
11.6 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 6.0, 65°C | Thermosphaera aggregans | |
53.9 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 5.0, 65°C | Thermosphaera aggregans | |
72.9 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant E387G, pH 6.5, 65°C, presence of 2 M formate | Thermosphaera aggregans | |
305 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 4.0, 65°C | Thermosphaera aggregans | |
309 | - |
2-nitrophenyl beta-D-glucopyranoside | wild-type, pH 6.5, 65°C | Thermosphaera aggregans | |
439.8 | - |
2-nitrophenyl beta-D-glucopyranoside | wild-type, pH 6.5, 65°C, presence of 2 M formate | Thermosphaera aggregans | |
970 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 3.0, 65°C | Thermosphaera aggregans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4.2 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 6.0, 65°C | Thermosphaera aggregans | |
17.2 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 5.0, 65°C | Thermosphaera aggregans | |
72.9 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant E387G, pH 6.5, 65°C, presence of 2 M formate | Thermosphaera aggregans | |
91 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 4.0, 65°C | Thermosphaera aggregans | |
147.4 | - |
2-nitrophenyl beta-D-glucopyranoside | mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 3.0, 65°C | Thermosphaera aggregans | |
1164 | - |
2-nitrophenyl beta-D-glucopyranoside | wild-type, pH 6.5, 65°C, presence of 2 M formate | Thermosphaera aggregans | |
1575 | - |
2-nitrophenyl beta-D-glucopyranoside | wild-type, pH 6.5, 65°C | Thermosphaera aggregans |