Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli BL21(DE3) | Cupriavidus necator |
gene kguK, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) pLysS | Cupriavidus necator |
General Stability | Organism |
---|---|
the enzyme is stabilized by storage in an imidazole solution which is removed just before use | Cupriavidus necator |
the enzyme is unstable in its pure form | Cupriavidus necator |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics of the recombinant enzyme | Cupriavidus necator | |
0.073 | - |
ATP | pH 8.0, temperature not specified in the publication | Cupriavidus necator | |
0.35 | - |
2-dehydro-D-gluconate | pH 8.0, temperature not specified in the publication | Cupriavidus necator | |
1.4 | - |
2-keto-3-deoxy-D-gluconate | pH 8.0, temperature not specified in the publication | Cupriavidus necator | |
4 | - |
2-keto-D-gulonate | pH 8.0, temperature not specified in the publication | Cupriavidus necator | |
25 | - |
2-keto-3-deoxy-D-gulonate | pH 8.0, temperature not specified in the publication | Cupriavidus necator |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Cupriavidus necator |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 2-dehydro-D-gluconate | Cupriavidus necator | - |
ADP + 6-phospho-2-dehydro-D-gluconate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cupriavidus necator | Q0K080 | - |
- |
Cupriavidus necator | Q0K080 | i.e. Ralstonia eutropha strain H16 | - |
Purification (Comment) | Organism |
---|---|
- |
Cupriavidus necator |
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) pLysS by nickel affinity chromatography and desalting gel filtration | Cupriavidus necator |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
8.7 | - |
purified recombinant His-tagged enzyme, pH 8.0, 37°C | Cupriavidus necator |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 2-dehydro-D-gluconate | - |
Cupriavidus necator | ADP + 6-phospho-2-dehydro-D-gluconate | - |
? | |
ATP + 2-dehydro-D-gluconate | - |
Cupriavidus necator | ADP + 6-phospho-2-dehydro-D-gluconate | - |
r | |
ATP + 2-dehydro-D-gluconate | preferred substrate | Cupriavidus necator | ADP + 6-phospho-2-dehydro-D-gluconate | - |
r | |
ATP + 2-dehydro-D-gulonate | - |
Cupriavidus necator | ADP + 6-phospho-2-dehydro-D-gulonate | - |
r | |
ATP + 2-keto-3-deoxy-D-gluconate | - |
Cupriavidus necator | ADP + ? | - |
? | |
ATP + 2-keto-3-deoxy-D-gulonate | - |
Cupriavidus necator | ADP + ? | - |
? | |
ATP + 2-keto-D-gulonate | - |
Cupriavidus necator | ADP + ? | - |
? | |
additional information | the substrate specificity analysis reveals that three ketoacids (2-dehydro-3-deoxy-D-gluconate, 2-dehydro-D-gulonate, and 2-dehydro-3-deoxy-D-gulonate) with structures close to the natural substrate (2-dehydro-D-gluconate) are successfully phosphorylated at an efficiency lower than or comparable to 2-dehydrogluconate, as depicted by the measured kinetic constant values. Assay method optimization, overview | Cupriavidus necator | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
? | x * 35700, recombinant His-tagged enzyme, SDS-PAGE | Cupriavidus necator |
Synonyms | Comment | Organism |
---|---|---|
2-dehydrogluconokinase | - |
Cupriavidus necator |
2-ketogluconate kinase | - |
Cupriavidus necator |
KGUK | - |
Cupriavidus necator |
KGUKCnec | - |
Cupriavidus necator |
More | see also EC 2.7.1.45 | Cupriavidus necator |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Cupriavidus necator |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.1 | - |
2-keto-D-gulonate | pH 8.0, temperature not specified in the publication | Cupriavidus necator | |
2.1 | - |
2-keto-3-deoxy-D-gulonate | pH 8.0, temperature not specified in the publication | Cupriavidus necator | |
5.71 | - |
ATP | pH 8.0, temperature not specified in the publication | Cupriavidus necator | |
5.8 | - |
2-dehydro-D-gluconate | pH 8.0, temperature not specified in the publication | Cupriavidus necator | |
10.2 | - |
2-keto-3-deoxy-D-gluconate | pH 8.0, temperature not specified in the publication | Cupriavidus necator |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Cupriavidus necator |
General Information | Comment | Organism |
---|---|---|
physiological function | 2-ketogluconate kinase (KGUK) is involved in the glucose and 2-ketogluconate catabolism of several aerobic bacteria, but relatively few bacterial species are able to utilize 2-ketogluconate as the sole carbon source for growth and energy provision | Cupriavidus necator |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.083 | - |
2-keto-3-deoxy-D-gulonate | pH 8.0, temperature not specified in the publication | Cupriavidus necator | |
0.246 | - |
2-keto-D-gulonate | pH 8.0, temperature not specified in the publication | Cupriavidus necator | |
7.37 | - |
2-keto-3-deoxy-D-gluconate | pH 8.0, temperature not specified in the publication | Cupriavidus necator | |
16.77 | - |
2-dehydro-D-gluconate | pH 8.0, temperature not specified in the publication | Cupriavidus necator | |
78.22 | - |
ATP | pH 8.0, temperature not specified in the publication | Cupriavidus necator |