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Literature summary for 2.5.1.55 extracted from

  • Cochrane, F.C.; Cookson, T.V.; Jameson, G.B.; Parker, E.J.
    Reversing evolution: re-establishing obligate metal ion dependence in a metal-independent KDO8P synthase (2009), J. Mol. Biol., 390, 646-661.
    View publication on PubMed
Search for more literature for 2.5.1.55

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21(DE3) Neisseria meningitidis

Crystallization (Commentary)

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

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

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 [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

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 [Da] Molecular Weight Maximum [Da] Comment Organism
30000
-
 monomer, SDS-PAGE Neisseria meningitidis
130000
-
 native protein, gel-filtration chromatography Neisseria meningitidis

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Neisseria meningitidis Q9JZ55
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Neisseria meningitidis

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

Subunits Comment Organism
tetramer 4 * 30000, gel-filtration chromatography Neisseria meningitidis

Synonyms

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 [°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 Range [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
 50 mM 1,3-bis[tris(hydroxymethyl)-methylamino]propane (BTP) buffer Neisseria meningitidis

pH Range

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 [mM/s]

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