Cloned (Comment) | Organism |
---|---|
expression with C-terminal His-tag | Bacillus subtilis |
Protein Variants | Comment | Organism |
---|---|---|
C110S | 0.4 mol iron per mol of protein, no enzymic activity | Bacillus subtilis |
C230S | 0.6 mol iron per mol of protein, no enzymic activity | Bacillus subtilis |
C259S | 4.5 mol iron per mol of protein, 80% of wild-type activity | Bacillus subtilis |
C318S | 3.3 mol iron per mol of protein, 75% of wild-type activity | Bacillus subtilis |
C318S/C320S | 0.3 mol iron per mol of protein, no enzymic activity | Bacillus subtilis |
C320S | 1.5 mol iron per mol of protein, no enzymic activity | Bacillus subtilis |
C82S | 4.3 mol iron per mol of protein, activity similar to wild-type | Bacillus subtilis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
O2 | complete loss of activity upon purification of enzyme in aerobic conditions or exposure to oxygen overnight | Bacillus subtilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.36 | - |
dihydroxyacetone phosphate | 25°C | Bacillus subtilis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Iron | enzyme contains a [4Fe-4S]-cluster, coordinated by residues C110, C230, C320. 3.8 mol iron per mol of enzyme | Bacillus subtilis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
41000 | - |
3 * 41000, calculated | Bacillus subtilis |
124000 | - |
gel filtration | Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | - |
isoform NadA | - |
Purification (Comment) | Organism |
---|---|
recombinant protein, complete loss of activity upon purification of enzyme in aerobic conditions or exposure to oxygen overnight | Bacillus subtilis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
discontinuous enzymatic assay that couples the production of iminoaspartate by NadB with the condensation between DHAP and iminoaspartate to form quinolinic acid catalyzed by NadA. The assay is linear up to 0.25 mg of NadA, 10 microg of NadB is the lowest amount suitable to measure NadA activity, and under anaerobic conditions, NadA activity becomes independent of fumarate concentration, starting from 1 mM fumarate, but decreases at concentrations higher than 2 mM fumarate, due to inhibition of NadB by fumarate | Bacillus subtilis |
0.027 | - |
25°C, presence of oxygen to reoxidize NadB in the coupled assay | Bacillus subtilis |
0.05 | - |
25°C, presence of fumarate as electron acceptor for NadB in the coupled assay | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dihydroxyacetone phosphate + iminoaspartate | - |
Bacillus subtilis | ? + H2O + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
trimer | 3 * 41000, calculated | Bacillus subtilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
iron-sulfur centre | enzyme contains a [4Fe-4S]-cluster, coordinated by residues C110, C230, C320. 3.8 mol iron per mol of enzyme, 3.3 mol sulfur per mol of enzyme | Bacillus subtilis |