Cloned (Comment) | Organism |
---|---|
- |
Saccharolobus solfataricus |
Crystallization (Comment) | Organism |
---|---|
hanging drop, vapor-diffusion method, crystal structure of the apo form of glucose dehydrogenase to a resolution of 1.8 A and a complex with its required cofactor, NADP+, to a resolution of 2.3 A. Complexes of the enzyme with D-glucose and D-xylose are presented to resolutions of 1.6 and 1.5 A. A T41A mutation is engineered to enable the trapping of substrate in the crystal | Saccharolobus solfataricus |
Protein Variants | Comment | Organism |
---|---|---|
T41A | kcat/Km value of the mutant enzyme for beta-D-glucose (with NAD+) is about 30fold lower compared to wild-type enzyme, kcat/Km value of the mutant enzyme for beta-D-flucose (with NADP+) is about 90fold lower compared to wild-type enzyme | Saccharolobus solfataricus |
T41V | kcat/Km value of the mutant enzyme for beta-D-glucose (with NAD+) is about 555fold lower compared to wild-type enzyme, kcat/Km value of the mutant enzyme for beta-D-flucose (with NADP+) is about 530fold lower compared to wild-type enzyme | Saccharolobus solfataricus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.18 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NADP+, wild-type enzyme | Saccharolobus solfataricus | |
0.25 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NAD+, wild-type enzyme | Saccharolobus solfataricus | |
0.44 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NADP+, wild-type enzyme | Saccharolobus solfataricus | |
0.57 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NAD+, wild-type enzyme | Saccharolobus solfataricus | |
1.3 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NADP+, wild-type enzyme | Saccharolobus solfataricus | |
1.5 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NAD+, wild-type enzyme | Saccharolobus solfataricus | |
20.4 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41A | Saccharolobus solfataricus | |
24.8 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41A | Saccharolobus solfataricus | |
29.2 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41A | Saccharolobus solfataricus | |
33.3 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41A | Saccharolobus solfataricus | |
59 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41V | Saccharolobus solfataricus | |
65.8 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41V | Saccharolobus solfataricus | |
72.5 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41V | Saccharolobus solfataricus | |
76.3 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41V | Saccharolobus solfataricus | |
109 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41A | Saccharolobus solfataricus | |
118 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41A | Saccharolobus solfataricus | |
175 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41V | Saccharolobus solfataricus | |
204 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41V | Saccharolobus solfataricus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharolobus solfataricus | O93715 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
beta-D-glucose + NAD+ + H2O | - |
Saccharolobus solfataricus | D-gluconate + NADH + 2 H+ | - |
ir | |
beta-D-glucose + NADP+ + H2O | - |
Saccharolobus solfataricus | D-gluconate + NADPH + 2 H+ | - |
ir | |
D-galactose + NAD+ + H2O | - |
Saccharolobus solfataricus | ? | - |
? | |
D-galactose + NADP+ + H2O | - |
Saccharolobus solfataricus | D-galactonate + NADPH + 2 H+ | - |
? | |
D-xylose + NAD+ + H2O | - |
Saccharolobus solfataricus | ? | - |
? | |
D-xylose + NADP+ + H2O | - |
Saccharolobus solfataricus | ? | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
assay at | Saccharolobus solfataricus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41V | Saccharolobus solfataricus | |
6 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41V | Saccharolobus solfataricus | |
7 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41V | Saccharolobus solfataricus | |
7 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41V | Saccharolobus solfataricus | |
8 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41V | Saccharolobus solfataricus | |
12 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41V | Saccharolobus solfataricus | |
14 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41A | Saccharolobus solfataricus | |
22 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41A | Saccharolobus solfataricus | |
22 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41A | Saccharolobus solfataricus | |
37 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NADP+, wild-type enzyme | Saccharolobus solfataricus | |
39 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41A | Saccharolobus solfataricus | |
44 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NADP+, wild-type enzyme | Saccharolobus solfataricus | |
48 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NADP+, wild-type enzyme | Saccharolobus solfataricus | |
56 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41A | Saccharolobus solfataricus | |
61 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NAD+, wild-type enzyme | Saccharolobus solfataricus | |
61 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NAD+, wild-type enzyme | Saccharolobus solfataricus | |
75 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NAD+, wild-type enzyme | Saccharolobus solfataricus | |
81 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41A | Saccharolobus solfataricus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Saccharolobus solfataricus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41V | Saccharolobus solfataricus | |
0.04 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41V | Saccharolobus solfataricus | |
0.07 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41V | Saccharolobus solfataricus | |
0.09 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41V | Saccharolobus solfataricus | |
0.11 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41V | Saccharolobus solfataricus | |
0.15 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41V | Saccharolobus solfataricus | |
0.2 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41A | Saccharolobus solfataricus | |
0.4 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41A | Saccharolobus solfataricus | |
0.5 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41A | Saccharolobus solfataricus | |
1.1 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NADP+, mutant enzyme T41A | Saccharolobus solfataricus | |
1.6 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41A | Saccharolobus solfataricus | |
2.8 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NAD+, mutant enzyme T41A | Saccharolobus solfataricus | |
37 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NADP+, wild-type enzyme | Saccharolobus solfataricus | |
50 | - |
beta-D-glucose | pH 7.5, 70°C, cosubstrate: NAD+, wild-type enzyme | Saccharolobus solfataricus | |
85 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NADP+, wild-type enzyme | Saccharolobus solfataricus | |
108 | - |
D-galactose | pH 7.5, 70°C, cosubstrate: NAD+, wild-type enzyme | Saccharolobus solfataricus | |
245 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NAD+, wild-type enzyme | Saccharolobus solfataricus | |
246 | - |
D-xylose | pH 7.5, 70°C, cosubstrate: NADP+, wild-type enzyme | Saccharolobus solfataricus |