Crystallization (Comment) | Organism |
---|---|
the three-dimensional structure of the H240N glucose 6-phosphate dehydrogenase is determined at 2.5 A resolution. Crystals are grown in the presence of either glucose 6-phosphate and NAD+ or glucose 6-phosphate and NADP+, hanging drop vapor diffusion method with 2.27 M ammonium sulfate | Leuconostoc mesenteroides |
Protein Variants | Comment | Organism |
---|---|---|
D177N | kcat/KM for D-glucose 6-phosphate (with cosubstrate NADP+) is fold lower than the value for the wild-type enzyme, kcat/KM for NADP+ is fold lower than the value for the wild-type enzyme, kcat/KM for D-glucose 6-phosphate (with cosubstrate NAD) is fold lower than the value for the wild-type enzyme, kcat/KM for D-glucose 6-phosphate (with cosubstrate NAD+) is fold lower than the value for the wild-type enzyme | Leuconostoc mesenteroides |
H178N | kcat/KM for D-glucose 6-phosphate (with cosubstrate NADP+) is fold lower than the value for the wild-type enzyme, kcat/KM for NADP+ is fold lower than the value for the wild-type enzyme, kcat/KM for D-glucose 6-phosphate (with cosubstrate NAD) is fold lower than the value for the wild-type enzyme, kcat/KM for D-glucose 6-phosphate (with cosubstrate NAD+) is fold lower than the value for the wild-type enzyme | Leuconostoc mesenteroides |
H250N | kcat/KM for D-glucose 6-phosphate (with cosubstrate NADP+) is fold lower than the value for the wild-type enzyme, kcat/KM for NADP+ is fold lower than the value for the wild-type enzyme, kcat/KM for D-glucose 6-phosphate (with cosubstrate NAD) is fold lower than the value for the wild-type enzyme, kcat/KM for D-glucose 6-phosphate (with cosubstrate NAD+) is fold lower than the value for the wild-type enzyme | Leuconostoc mesenteroides |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Leuconostoc mesenteroides |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0082 | - |
NADP+ | 25°C, pH 7.6, mutant enzyme D177N | Leuconostoc mesenteroides | |
0.0088 | - |
NADP+ | 25°C, pH 7.6, mutant enzyme H240N | Leuconostoc mesenteroides | |
0.0152 | - |
NADP+ | 25°C, pH 7.6, mutant enzyme H1787N | Leuconostoc mesenteroides | |
0.117 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme D177N | Leuconostoc mesenteroides | |
0.29 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme D177N | Leuconostoc mesenteroides | |
0.8 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme H240N | Leuconostoc mesenteroides | |
0.81 | - |
NAD+ | 25°C, pH 7.6, mutant enzyme H240N | Leuconostoc mesenteroides | |
0.95 | - |
NAD+ | 25°C, pH 7.6, mutant enzyme H1787N | Leuconostoc mesenteroides | |
1.14 | - |
NAD+ | 25°C, pH 7.6, mutant enzyme D177N | Leuconostoc mesenteroides | |
1.28 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme H240N | Leuconostoc mesenteroides | |
13.6 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme H1787N | Leuconostoc mesenteroides | |
42.9 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme H1787N | Leuconostoc mesenteroides |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glucose 6-phosphate + NAD+ | Leuconostoc mesenteroides | - |
6-phospho-D-glucono-1,5-lactone + NADH + H+ | - |
? | |
D-glucose 6-phosphate + NADP+ | Leuconostoc mesenteroides | - |
6-phospho-D-glucono-1,5-lactone + NADPH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Leuconostoc mesenteroides | - |
- |
- |
Purification (Comment) | Organism |
---|---|
mutant enzymes H250N, D177N and H178N | Leuconostoc mesenteroides |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
D-glucose 6-phosphate + NAD(P)+ = 6-phospho-D-glucono-1,5-lactone + NAD(P)H + H+ | mechanism in which His240 acts as the general base that abstracts the proton from the C1-hydroxyl group of glucose 6-phosphate, and the carboxylate group of D177 stabilizes the positive charge that forms on H240 in the transition state. The results also confirm the postulated role of His178 in binding the phosphate moiety of glucose 6-phosphate | Leuconostoc mesenteroides |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glucose 6-phosphate + NAD+ | - |
Leuconostoc mesenteroides | 6-phospho-D-glucono-1,5-lactone + NADH + H+ | - |
? | |
D-glucose 6-phosphate + NADP+ | - |
Leuconostoc mesenteroides | 6-phospho-D-glucono-1,5-lactone + NADPH + H+ | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Leuconostoc mesenteroides |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.007 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme H240N | Leuconostoc mesenteroides | |
0.007 | - |
NADP+ | 25°C, pH 7.6, mutant enzyme H240N | Leuconostoc mesenteroides | |
0.02 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme H240N | Leuconostoc mesenteroides | |
0.02 | - |
NAD+ | 25°C, pH 7.6, mutant enzyme H240N | Leuconostoc mesenteroides | |
1.1 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme D177N | Leuconostoc mesenteroides | |
1.1 | - |
NADP+ | 25°C, pH 7.6, mutant enzyme D177N | Leuconostoc mesenteroides | |
4.85 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme D177N | Leuconostoc mesenteroides | |
4.85 | - |
NAD+ | 25°C, pH 7.6, mutant enzyme D177N | Leuconostoc mesenteroides | |
13.8 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme H1787N | Leuconostoc mesenteroides | |
13.8 | - |
NADP+ | 25°C, pH 7.6, mutant enzyme H1787N | Leuconostoc mesenteroides | |
14.2 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme H1787N | Leuconostoc mesenteroides | |
14.2 | - |
NAD+ | 25°C, pH 7.6, mutant enzyme H1787N | Leuconostoc mesenteroides |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at | Leuconostoc mesenteroides |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0037 | - |
NADP+ | 25°C, pH 7.6, mutant enzyme H1787N | Leuconostoc mesenteroides | |
0.015 | - |
NADP+ | 25°C, pH 7.6, mutant enzyme D177N | Leuconostoc mesenteroides |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.005 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme H240N | Leuconostoc mesenteroides | |
0.025 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme H240N | Leuconostoc mesenteroides | |
0.025 | - |
NAD+ | 25°C, pH 7.6, mutant enzyme H240N | Leuconostoc mesenteroides | |
0.32 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme H1787N | Leuconostoc mesenteroides | |
0.8 | - |
NADP+ | 25°C, pH 7.6, mutant enzyme H240N | Leuconostoc mesenteroides | |
1.04 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme H1787N | Leuconostoc mesenteroides | |
4.25 | - |
NAD+ | 25°C, pH 7.6, mutant enzyme D177N | Leuconostoc mesenteroides | |
9.4 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme D177N | Leuconostoc mesenteroides | |
14.95 | - |
NAD+ | 25°C, pH 7.6, mutant enzyme H1787N | Leuconostoc mesenteroides | |
16.7 | - |
D-glucose 6-phosphate | 25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme D177N | Leuconostoc mesenteroides | |
134.1 | - |
NADP+ | 25°C, pH 7.6, mutant enzyme D177N | Leuconostoc mesenteroides | |
904 | - |
NADP+ | 25°C, pH 7.6, mutant enzyme H1787N | Leuconostoc mesenteroides |