Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Rosetta2 (DE3) cells | Sulfurisphaera tokodaii |
expression in Escherichia coli | Sulfurisphaera tokodaii |
Protein Variants | Comment | Organism |
---|---|---|
I189V | the mutant has decreased kcat and kcat/Km values at both 25°C and 80°C compared to the wild type enzyme | Sulfurisphaera tokodaii |
V254I | mutant has improved kcat and kcat/Km values at both 25°C and 80°C. The thermal stability of the mutant enzyme was comparable to that of the wild-type enzyme. Calculation of the energetic contribution of the V254I mutation for the dehydrogenase reaction reveals that the mutation destabilizes the enzyme-NADPD-glucose ternary complex and reduces the transition-state energy, thus enhancing catalysis | Sulfurisphaera tokodaii |
V254I | the mutant has improved kcat and kcat/Km values at both 25°C and 80°C compared to the wild type enzyme | Sulfurisphaera tokodaii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00588 | - |
NADP+ | pH 9.0, 25°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.00588 | - |
NADP+ | wild type enzyme, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.00876 | - |
NADP+ | pH 9.0, 25°C, mutant enzyme V254I | Sulfurisphaera tokodaii | |
0.00876 | - |
NADP+ | mutant enzyme V254I, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.00927 | - |
NADP+ | pH 9.0, 25°C, mutant enzyme I189V | Sulfurisphaera tokodaii | |
0.00927 | - |
NADP+ | mutant enzyme I189V, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.0286 | - |
D-glucose | pH 9.0, 25°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.0286 | - |
D-glucose | wild type enzyme, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.0293 | - |
D-glucose | pH 9.0, 25°C, mutant enzyme I189V | Sulfurisphaera tokodaii | |
0.0293 | - |
D-glucose | mutant enzyme I189V, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.0391 | - |
D-glucose | pH 9.0, 25°C, mutant enzyme V254I | Sulfurisphaera tokodaii | |
0.0391 | - |
D-glucose | mutant enzyme V254I, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.109 | - |
NADP+ | pH 9.0, 80°C, mutant enzyme I189V | Sulfurisphaera tokodaii | |
0.109 | - |
NADP+ | mutant enzyme I189V, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.117 | - |
NADP+ | pH 9.0, 80°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.118 | - |
NADP+ | wild type enzyme, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.132 | - |
NADP+ | pH 9.0, 80°C, mutant enzyme V254I | Sulfurisphaera tokodaii | |
0.132 | - |
NADP+ | mutant enzyme V254I, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.287 | - |
D-glucose | pH 9.0, 80°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.287 | - |
D-glucose | wild type enzyme, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.314 | - |
D-glucose | pH 9.0, 80°C, mutant enzyme I189V | Sulfurisphaera tokodaii | |
0.314 | - |
D-glucose | mutant enzyme I189V, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.457 | - |
D-glucose | pH 9.0, 80°C, mutant enzyme V254I | Sulfurisphaera tokodaii | |
0.457 | - |
D-glucose | mutant enzyme V254I, at pH 9.0 and 80°C | Sulfurisphaera tokodaii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glucose + NADP+ | Sulfurisphaera tokodaii | - |
D-glucono-1,5-lactone + NADPH + H+ | - |
? | |
D-glucose + NADP+ | Sulfurisphaera tokodaii DSM 16993 | - |
D-glucono-1,5-lactone + NADPH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sulfurisphaera tokodaii | Q96ZY1 | - |
- |
Sulfurisphaera tokodaii | Q96ZY7 | - |
- |
Sulfurisphaera tokodaii DSM 16993 | Q96ZY7 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Sulfurisphaera tokodaii |
HiTrapQ column chromatography and Resource-phenyl column chromatography | Sulfurisphaera tokodaii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glucose + NADP+ | - |
Sulfurisphaera tokodaii | D-glucono-1,5-lactone + NADPH + H+ | - |
? | |
D-glucose + NADP+ | the enzyme prefers NADP+ over NAD+ as the cofactor. It shows a larger Km value for NAD+ (4.5 mM) than for NADP+ (0.12 mM) at 80°C. At 25°C the Km values are 0.069 mM for NAD+ and 0.0059 mM for NADP+ | Sulfurisphaera tokodaii | D-glucono-1,5-lactone + NADPH + H+ | - |
? | |
D-glucose + NADP+ | - |
Sulfurisphaera tokodaii DSM 16993 | D-glucono-1,5-lactone + NADPH + H+ | - |
? | |
D-glucose + NADP+ | the enzyme prefers NADP+ over NAD+ as the cofactor. It shows a larger Km value for NAD+ (4.5 mM) than for NADP+ (0.12 mM) at 80°C. At 25°C the Km values are 0.069 mM for NAD+ and 0.0059 mM for NADP+ | Sulfurisphaera tokodaii DSM 16993 | D-glucono-1,5-lactone + NADPH + H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
GDH | - |
Sulfurisphaera tokodaii |
glucose-1-dehydrogenase | - |
Sulfurisphaera tokodaii |
NAD(P)-dependent GDH | - |
Sulfurisphaera tokodaii |
nicotinamide adenine dinucleotide (phosphate)-dependent glucose-1-dehydrogenase | - |
Sulfurisphaera tokodaii |
STK_17040 | locus name | Sulfurisphaera tokodaii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0097 | - |
NADP+ | mutant enzyme I189V, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.0097 | - |
D-glucose | mutant enzyme I189V, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.0101 | - |
NADP+ | wild type enzyme, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.0101 | - |
D-glucose | wild type enzyme, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.0167 | - |
NADP+ | mutant enzyme V254I, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.0167 | - |
D-glucose | mutant enzyme V254I, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.085 | - |
NADP+ | pH 9.0, 25°C, mutant enzyme I189V | Sulfurisphaera tokodaii | |
0.085 | - |
D-glucose | pH 9.0, 25°C, mutant enzyme I189V | Sulfurisphaera tokodaii | |
0.085 | - |
D-glucose | mutant enzyme I189V, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.0857 | - |
NADP+ | mutant enzyme I189V, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.11 | - |
NADP+ | pH 9.0, 25°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.11 | - |
D-glucose | pH 9.0, 25°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.11 | - |
NADP+ | wild type enzyme, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.11 | - |
D-glucose | wild type enzyme, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.205 | - |
NADP+ | pH 9.0, 25°C, mutant enzyme V254I | Sulfurisphaera tokodaii | |
0.205 | - |
D-glucose | pH 9.0, 25°C, mutant enzyme V254I | Sulfurisphaera tokodaii | |
0.205 | - |
NADP+ | mutant enzyme V254I, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.205 | - |
D-glucose | mutant enzyme V254I, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
9.7 | - |
NADP+ | pH 9.0, 80°C, mutant enzyme I189V | Sulfurisphaera tokodaii | |
9.7 | - |
D-glucose | pH 9.0, 80°C, mutant enzyme I189V | Sulfurisphaera tokodaii | |
10.1 | - |
NADP+ | pH 9.0, 80°C, wild-type enzyme | Sulfurisphaera tokodaii | |
10.1 | - |
D-glucose | pH 9.0, 80°C, wild-type enzyme | Sulfurisphaera tokodaii | |
16.7 | - |
NADP+ | pH 9.0, 80°C, mutant enzyme V254I | Sulfurisphaera tokodaii | |
16.7 | - |
D-glucose | pH 9.0, 80°C, mutant enzyme V254I | Sulfurisphaera tokodaii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
assay at | Sulfurisphaera tokodaii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Sulfurisphaera tokodaii | |
NADP+ | the enzyme prefers NADP+ over NAD+ as the cofactor. It shows a larger Km value for NAD+ (4.5 mM) than for NADP+ (0.12 mM) at 80°C. At 25°C the Km values are 0.069 mM for NAD+ and 0.0059 mM for NADP+ | Sulfurisphaera tokodaii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.092 | - |
NADP+ | pH 9.0, 25°C, mutant enzyme I189V | Sulfurisphaera tokodaii | |
0.092 | - |
NADP+ | mutant enzyme I189V, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.126 | - |
NADP+ | pH 9.0, 80°C, mutant enzyme V254I | Sulfurisphaera tokodaii | |
0.126 | - |
NADP+ | mutant enzyme V254I, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.186 | - |
NADP+ | pH 9.0, 25°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.186 | - |
NADP+ | wild type enzyme, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.234 | - |
NADP+ | pH 9.0, 25°C, mutant enzyme V254I | Sulfurisphaera tokodaii | |
0.234 | - |
NADP+ | mutant enzyme V254I, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
0.31 | - |
D-glucose | pH 9.0, 80°C, mutant enzyme I189V | Sulfurisphaera tokodaii | |
0.31 | - |
D-glucose | mutant enzyme I189V, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.351 | - |
D-glucose | pH 9.0, 80°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.351 | - |
D-glucose | wild type enzyme, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.365 | - |
D-glucose | pH 9.0, 80°C, mutant enzyme V254I | Sulfurisphaera tokodaii | |
0.365 | - |
D-glucose | mutant enzyme V254I, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.863 | - |
NADP+ | pH 9.0, 80°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.863 | - |
NADP+ | wild type enzyme, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
0.893 | - |
NADP+ | pH 9.0, 80°C, mutant enzyme I189V | Sulfurisphaera tokodaii | |
0.893 | - |
NADP+ | mutant enzyme I189V, at pH 9.0 and 80°C | Sulfurisphaera tokodaii | |
2.91 | - |
D-glucose | pH 9.0, 25°C, mutant enzyme I189V | Sulfurisphaera tokodaii | |
2.91 | - |
D-glucose | mutant enzyme I189V, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
3.82 | - |
D-glucose | pH 9.0, 25°C, wild-type enzyme | Sulfurisphaera tokodaii | |
3.82 | - |
D-glucose | wild type enzyme, at pH 9.0 and 25°C | Sulfurisphaera tokodaii | |
5.25 | - |
D-glucose | pH 9.0, 25°C, mutant enzyme V254I | Sulfurisphaera tokodaii | |
5.25 | - |
D-glucose | mutant enzyme V254I, at pH 9.0 and 25°C | Sulfurisphaera tokodaii |