Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli BL21(DE3) | Neisseria meningitidis |
Crystallization (Comment) | Organism |
---|---|
vapor-diffusion hanging drop method, pH 4.6 and pH 5.0, no diffraction quality crystals at higher pH in the range of the active enzyme | Neisseria meningitidis |
Protein Variants | Comment | Organism |
---|---|---|
additional information | replacement of asparagine with metal-binding cysteine is not sufficient to produce a metal-dependent mutant, 3 to 4 mutations establish a fully functional obligate metal-dependent KDO8PS, metal-dependent mutants containing the C246S exchange are activated in the presence of Mn2+ and Cd2+ to specific activities comparable to the wild type, other metal-dependent mutations show lower activation levels, high concentrations can have inhibitory effects, Mg2+, Ca2+, Ni2+, Sr2+, and Ba2+ have little or no effect on activity, Co2+ and Fe2+ have varying effects, Zn2+ is inhibitory for all cloned enzymes | Neisseria meningitidis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cd2+ | inhibition of metal-independent wild type and mutant enzymes D247E, P249A at 2 microM and above, inhibition of mutant 247E/P249A at 8 microM and above | Neisseria meningitidis | |
Cu2+ | slight reduction of wild type enzyme activity | Neisseria meningitidis | |
Fe2+ | inhibitory for N23C, N23C/C246S/P249A, N23C/C246S/D247E/P249A | Neisseria meningitidis | |
Mn2+ | 15% reduced activity above 200 microM for mutant N23C/D247E/P249A | Neisseria meningitidis | |
Zn2+ | inhibitory for all cloned enzymes | Neisseria meningitidis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | N23C/C246S/D247E mutant control is barely active, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
additional information | - |
additional information | N23C/C246S/D247E/P249A mutant control is barely active, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
additional information | - |
additional information | N23C/C246S/P249A mutant control is barely active, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.0024 | - |
phosphoenolpyruvate | N23C/C247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.0025 | - |
phosphoenolpyruvate | wild type, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.0027 | - |
phosphoenolpyruvate | N23C/C247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.0028 | - |
phosphoenolpyruvate | N23C mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.0033 | - |
phosphoenolpyruvate | N23C/P249A mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.0038 | - |
phosphoenolpyruvate | N23C/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.0047 | - |
phosphoenolpyruvate | P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.0049 | - |
phosphoenolpyruvate | D247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.0054 | - |
phosphoenolpyruvate | N23C mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.0057 | - |
D-arabinose 5-phosphate | wild type, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.0066 | - |
phosphoenolpyruvate | N23C/C246S mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.0077 | - |
phosphoenolpyruvate | N23C/C247E mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.0085 | - |
phosphoenolpyruvate | D247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.0093 | - |
phosphoenolpyruvate | C246S mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.0095 | - |
phosphoenolpyruvate | N23C/C246S/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.012 | - |
D-arabinose 5-phosphate | N23C mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.012 | - |
D-arabinose 5-phosphate | P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.013 | - |
D-arabinose 5-phosphate | N23C/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.0153 | - |
phosphoenolpyruvate | N23C/C246S/D247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.017 | - |
phosphoenolpyruvate | N23C/C247E/P249A mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.022 | - |
phosphoenolpyruvate | N23C/C246S/D247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.05 | - |
phosphoenolpyruvate | N23C/C246S mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.06 | - |
D-arabinose 5-phosphate | D247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.06 | - |
D-arabinose 5-phosphate | N23C mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.074 | - |
D-arabinose 5-phosphate | D247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.097 | - |
D-arabinose 5-phosphate | N23C/C246S mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.107 | - |
D-arabinose 5-phosphate | N23C/C247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.109 | - |
D-arabinose 5-phosphate | N23C/P249A mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.14 | - |
D-arabinose 5-phosphate | N23C/C247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.19 | - |
D-arabinose 5-phosphate | N23C/C246S/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.213 | - |
D-arabinose 5-phosphate | C246S mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.24 | - |
D-arabinose 5-phosphate | N23C/C247E mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.33 | - |
D-arabinose 5-phosphate | N23C/C247E/P249A mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.498 | - |
D-arabinose 5-phosphate | N23C/C246S/D247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
0.54 | - |
D-arabinose 5-phosphate | N23C/C246S/D247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
1.04 | - |
D-arabinose 5-phosphate | N23C/C246S mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ba2+ | no activation of wild type enzyme and metal-dependent mutants, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4 | Neisseria meningitidis | |
Ca2+ | no activation of wild type enzyme and metal-dependent mutants, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4 | Neisseria meningitidis | |
Cd2+ | activation of metal-dependent mutants, inhibition of wild type and metal-independent mutants, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4 | Neisseria meningitidis | |
Co2+ | no activation of wild type enzyme, adverse effect on mutant N23C, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4 | Neisseria meningitidis | |
Cu2+ | no activation of mutant enzymes containing the N23C substitution, slight reduction of wild type enzyme activity, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4 | Neisseria meningitidis | |
Fe2+ | no activation of wild type enzyme and metal-independent mutants, activation of N23C/D247E, N23C/C246S/D247E, inhibitory for N23C, N23C/C246S/P249A, N23C/C246S/D247E/P249A, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4 | Neisseria meningitidis | |
Mg2+ | no activation of wild type enzyme and metal-dependent mutants, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4 | Neisseria meningitidis | |
Mn2+ | no activation of wild type enzyme, activation of metal-dependent mutants, 15% reduced activity above 200 microM for mutant N23C/D247E/P249A, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4 | Neisseria meningitidis | |
additional information | the metal chelator EDTA has no effect on the activity of the metal-independent wild type enzyme and on the metal-independent mutants C246S, D247E, D247E/P249A or P249A | Neisseria meningitidis | |
Ni2+ | no activation of wild type enzyme and metal-dependent mutants, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4 | Neisseria meningitidis | |
Sr2+ | no activation of wild type enzyme and metal-dependent mutants, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4 | Neisseria meningitidis | |
Zn2+ | no activation of wild type enzyme, inhibitory for all cloned enzymes, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4 | Neisseria meningitidis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
30000 | - |
monomer, SDS-PAGE | Neisseria meningitidis |
130000 | - |
native protein, gel-filtration chromatography | Neisseria meningitidis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
phosphoenolpyruvate + D-arabinose 5-phosphate + H2O | Neisseria meningitidis | - |
2-dehydro-3-deoxy-D-octonate 8-phosphate + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Neisseria meningitidis | Q9JZ55 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Neisseria meningitidis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.01 | - |
N23C/C246S/D247E mutant control, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
0.01 | - |
N23C/C246S/D247E/P249A mutant control, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
0.06 | - |
N23C/C246S/P249A mutant control, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
0.19 | - |
N23C/C246S mutant control, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
0.3 | - |
N23C/C247E mutant control, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
0.35 | - |
N23C/C247E/P249A mutant control, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
0.76 | - |
N23C/P249A mutant control, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
1.9 | - |
N23C mutant control, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
2.34 | - |
N23C/P249A mutant, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis |
2.9 | - |
N23C mutant, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis |
3.1 | - |
N23C/C247E mutant, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis |
4.34 | - |
N23C/C247E/P249A mutant, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis |
5 | - |
N23C/C246S mutant, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis |
5.64 | - |
N23C/C246S/D247E/P249A mutant, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis |
6.3 | - |
D247E/P249A mutant, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
6.39 | - |
D247E mutant, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
8.4 | - |
N23C/C246S/D247E mutant, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis |
9.8 | - |
N23C/C246S/P249A mutant, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis |
10.1 | - |
C246S mutant, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
11.7 | - |
P249A mutant, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
12 | - |
wild type, 200 microM phosphoenolpyruvate, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
phosphoenolpyruvate + D-arabinose 5-phosphate + H2O | - |
Neisseria meningitidis | 2-dehydro-3-deoxy-D-octonate 8-phosphate + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | 4 * 30000, gel-filtration chromatography | Neisseria meningitidis |
Synonyms | Comment | Organism |
---|---|---|
metal-independent 3-deoxy-D-manno-octulosonate 8-phosphate synthase | - |
Neisseria meningitidis |
metal-independent KDO8PS | - |
Neisseria meningitidis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.5 | Neisseria meningitidis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | 70 | 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.5, 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 60°C, 70°C | Neisseria meningitidis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | N23C/C246S/D247E mutant control is barely active, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
additional information | - |
additional information | N23C/C246S/D247E/P249A mutant control is barely active, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
additional information | - |
additional information | N23C/C246S/P249A mutant control is barely active, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.02 | - |
phosphoenolpyruvate | N23C/C246S mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.31 | - |
phosphoenolpyruvate | N23C/C247E/P249A mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.38 | - |
phosphoenolpyruvate | N23C/C247E mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.38 | - |
phosphoenolpyruvate | N23C/P249A mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.93 | - |
phosphoenolpyruvate | N23C mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.94 | - |
phosphoenolpyruvate | N23C/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
1.42 | - |
phosphoenolpyruvate | N23C mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
2.02 | - |
phosphoenolpyruvate | N23C/C247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
2.71 | - |
phosphoenolpyruvate | N23C/C246S mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
3.27 | - |
phosphoenolpyruvate | N23C/C247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
3.6 | - |
phosphoenolpyruvate | D247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
3.9 | - |
phosphoenolpyruvate | N23C/C246S/D247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
4.4 | - |
phosphoenolpyruvate | D247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
4.4 | - |
phosphoenolpyruvate | N23C/C246S/D247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
4.79 | - |
phosphoenolpyruvate | N23C/C246S/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
5.5 | - |
phosphoenolpyruvate | C246S mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
8 | - |
phosphoenolpyruvate | wild type, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
8.6 | - |
phosphoenolpyruvate | P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer | Neisseria meningitidis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
3.8 | 9.15 | 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, wild type enzyme is most active between pH 6.8 and pH 8.1 | Neisseria meningitidis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | N23C/C246S/D247E mutant control is barely active, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
additional information | - |
additional information | N23C/C246S/D247E/P249A mutant control is barely active, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
additional information | - |
additional information | N23C/C246S/P249A mutant control is barely active, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.02 | - |
D-arabinose 5-phosphate | N23C/C246S mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.4 | - |
phosphoenolpyruvate | N23C/C246S mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
0.9 | - |
D-arabinose 5-phosphate | N23C/C247E/P249A mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
1.6 | - |
D-arabinose 5-phosphate | N23C/C247E mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
3.5 | - |
D-arabinose 5-phosphate | N23C/P249A mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
7.9 | - |
D-arabinose 5-phosphate | N23C/C246S/D247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
8.1 | - |
D-arabinose 5-phosphate | N23C/C246S/D247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
16 | - |
D-arabinose 5-phosphate | N23C mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
18 | - |
phosphoenolpyruvate | N23C/C247E/P249A mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
19 | - |
D-arabinose 5-phosphate | N23C/C247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
23 | - |
D-arabinose 5-phosphate | N23C/C247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
25 | - |
D-arabinose 5-phosphate | N23C/C246S/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
26 | - |
D-arabinose 5-phosphate | C246S mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
28 | - |
D-arabinose 5-phosphate | N23C/C246S mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
49 | - |
D-arabinose 5-phosphate | D247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
49 | - |
phosphoenolpyruvate | N23C/C247E mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
72 | - |
D-arabinose 5-phosphate | N23C/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
73 | - |
D-arabinose 5-phosphate | D247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
115 | - |
phosphoenolpyruvate | N23C/P249A mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
118 | - |
D-arabinose 5-phosphate | N23C mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
200 | - |
phosphoenolpyruvate | N23C/C246S/D247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
247 | - |
phosphoenolpyruvate | N23C/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
257 | - |
phosphoenolpyruvate | N23C/C246S/D247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
263 | - |
phosphoenolpyruvate | N23C mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
332 | - |
phosphoenolpyruvate | N23C mutant control, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
411 | - |
phosphoenolpyruvate | N23C/C246S mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
424 | - |
phosphoenolpyruvate | D247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
504 | - |
phosphoenolpyruvate | N23C/C246S/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
591 | - |
phosphoenolpyruvate | C246S mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
717 | - |
D-arabinose 5-phosphate | P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
748 | - |
phosphoenolpyruvate | N23C/C247E/P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
898 | - |
phosphoenolpyruvate | D247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
1363 | - |
phosphoenolpyruvate | N23C/C247E mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 90 microM MnSO4.H2O | Neisseria meningitidis | |
1400 | - |
D-arabinose 5-phosphate | wild type, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
1830 | - |
phosphoenolpyruvate | P249A mutant, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis | |
3200 | - |
phosphoenolpyruvate | wild type, 37°C, 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer, pH 7.4, 20 microM EDTA | Neisseria meningitidis |