EC Number | Cloned (Comment) | Organism |
---|---|---|
6.3.1.5 | nadE gene, expression of the enzyme as a His-tagged protein and as a thioredoxin-His6-fusion protein in insect cells of Thrichoplusia ni BTI-TN-5B1-4 via baculovirus infection, for large scale production, and in Escherichia coli | Mycobacterium tuberculosis |
6.3.5.1 | most efficient expression of His-tagged NadE-738 and NadE-679 in Escherichia coli strain Origami | Mycobacterium tuberculosis |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
6.3.1.5 | thrombin | enzyme is highly susceptible too proteolytic cleavage | Mycobacterium tuberculosis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
6.3.1.5 | 0.00052 | - |
deamido-NAD+ | pH 8.5, 37°C, recombinant truncated enzyme | Mycobacterium tuberculosis | |
6.3.1.5 | 0.00079 | - |
ATP | pH 8.5, 37°C, recombinant truncated enzyme | Mycobacterium tuberculosis | |
6.3.1.5 | 0.0011 | - |
glutamine | pH 8.5, 37°C, recombinant full length enzyme | Mycobacterium tuberculosis | |
6.3.1.5 | 0.0013 | - |
ATP | pH 8.5, 37°C, recombinant full length enzyme | Mycobacterium tuberculosis | |
6.3.1.5 | 0.0016 | - |
glutamine | pH 8.5, 37°C, recombinant truncated enzyme | Mycobacterium tuberculosis | |
6.3.1.5 | 0.0029 | - |
deamido-NAD+ | pH 8.5, 37°C, recombinant full length enzyme | Mycobacterium tuberculosis | |
6.3.1.5 | 0.025 | - |
NH4+ | pH 8.5, 37°C, recombinant truncated enzyme | Mycobacterium tuberculosis | |
6.3.1.5 | 0.027 | - |
NH4+ | pH 8.5, 37°C, recombinant full length enzyme | Mycobacterium tuberculosis | |
6.3.5.1 | 0.52 | - |
deamido-NAD+ | 37°C, pH 8.0, recombinant NadE-679 | Mycobacterium tuberculosis | |
6.3.5.1 | 0.79 | - |
ATP | 37°C, pH 8.0, recombinant NadE-679 | Mycobacterium tuberculosis | |
6.3.5.1 | 1.1 | - |
L-glutamine | 37°C, pH 8.8, recombinant NadE-738 | Mycobacterium tuberculosis | |
6.3.5.1 | 1.3 | - |
ATP | 37°C, pH 8.5, recombinant NadE-738 | Mycobacterium tuberculosis | |
6.3.5.1 | 1.6 | - |
L-glutamine | 37°C, pH 8.5, recombinant NadE-679 | Mycobacterium tuberculosis | |
6.3.5.1 | 2.5 | - |
NH4+ | 37°C, pH 8.0, recombinant NadE-679 | Mycobacterium tuberculosis | |
6.3.5.1 | 2.9 | - |
deamido-NAD+ | 37°C, pH 8.5, recombinant NadE-738 | Mycobacterium tuberculosis | |
6.3.5.1 | 27 | - |
NH4+ | 37°C, pH 8.5, recombinant NadE-738 | Mycobacterium tuberculosis |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
6.3.1.5 | Mg2+ | required | Mycobacterium tuberculosis |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
6.3.5.1 | 67000 | - |
8 * 67000, SDS-PAGE | Mycobacterium tuberculosis |
6.3.5.1 | 600000 | - |
gel filtration, NadE-679 | Mycobacterium tuberculosis |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
6.3.1.5 | ATP + deamido-NAD+ + NH4+ | Mycobacterium tuberculosis | - |
AMP + diphosphate + NAD+ + H+ | - |
? | |
6.3.5.1 | ATP + deamido-NAD+ + L-glutamine + H2O | Mycobacterium tuberculosis | - |
AMP + diphosphate + NAD+ + L-glutamate | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
6.3.1.5 | Mycobacterium tuberculosis | - |
- |
- |
6.3.5.1 | Mycobacterium tuberculosis | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
6.3.1.5 | recombinant His-tagged enzyme and recombinant thioredoxin-His6-fusion protein from Escherichia coli and insect cells, the His-tag is cleaved off afterwards | Mycobacterium tuberculosis |
6.3.5.1 | recombinant NadE-738 and NadE-679, Ni2+-nitrilotriacetic acid agarose, gel filtration | Mycobacterium tuberculosis |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
6.3.1.5 | 0.009 | - |
partially purified recombinant His-tagged enzyme expressed in insect cells, substrate NH4+ | Mycobacterium tuberculosis |
6.3.1.5 | 0.033 | - |
purified recombinant HIs-tagged enzyme expressed in Escherichia coli, substrate glutamine | Mycobacterium tuberculosis |
6.3.1.5 | 0.1 | - |
purified recombinant His-tagged enzyme expressed in Escherichia coli, substrate NH4+ | Mycobacterium tuberculosis |
6.3.1.5 | 0.67 | - |
purified recombinant His-tagged truncated enzyme expressed in Escherichia coli, substrate NH4+ | Mycobacterium tuberculosis |
6.3.5.1 | 0.033 | - |
recombinant NadE-738, nitrogen donor glutamine | Mycobacterium tuberculosis |
6.3.5.1 | 0.1 | - |
recombinant NadE-738, nitrogen donor NH4+ | Mycobacterium tuberculosis |
6.3.5.1 | 0.44 | - |
recombinant NadE-679, nitrogen donor glutamine | Mycobacterium tuberculosis |
6.3.5.1 | 0.67 | - |
recombinant NadE-679, nitrogen donor NH4+ | Mycobacterium tuberculosis |
EC Number | Storage Stability | Organism |
---|---|---|
6.3.1.5 | -20°C, recombinant truncated enzyme, overnight, loss of 59% activity | Mycobacterium tuberculosis |
6.3.1.5 | -70°C, recombinant truncated enzyme, overnight, loss of 30% activity | Mycobacterium tuberculosis |
6.3.1.5 | -80 - 4°C, recombinant full length enzyme, several months | Mycobacterium tuberculosis |
6.3.1.5 | 25°C, recombinant truncated enzyme, stable | Mycobacterium tuberculosis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
6.3.1.5 | ATP + deamido-NAD+ + glutamine | - |
Mycobacterium tuberculosis | AMP + diphosphate + NAD+ + glutamate | - |
? | |
6.3.1.5 | ATP + deamido-NAD+ + NH4+ | - |
Mycobacterium tuberculosis | AMP + diphosphate + NAD+ + H+ | - |
? | |
6.3.5.1 | ATP + deamido-NAD+ + L-glutamine + H2O | - |
Mycobacterium tuberculosis | AMP + diphosphate + NAD+ + L-glutamate | - |
? | |
6.3.5.1 | ATP + deamido-NAD+ + NH4+ + H2O | - |
Mycobacterium tuberculosis | AMP + diphosphate + NAD+ | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
6.3.5.1 | octamer | 8 * 67000, SDS-PAGE | Mycobacterium tuberculosis |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
6.3.1.5 | NAD synthase | - |
Mycobacterium tuberculosis |
6.3.1.5 | NAD synthetase | - |
Mycobacterium tuberculosis |
6.3.1.5 | Nicotinamide adenine dinucleotide synthetase | - |
Mycobacterium tuberculosis |
6.3.5.1 | NadE-679 | short NadE form, probably raising from translation of NadE gene from an internal start codon | Mycobacterium tuberculosis |
6.3.5.1 | NadE-679 | - |
Mycobacterium tuberculosis |
6.3.5.1 | NadE-738 | - |
Mycobacterium tuberculosis |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
6.3.1.5 | 37 | - |
assay at | Mycobacterium tuberculosis |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
6.3.1.5 | 8.5 | - |
with substrate NH4+ | Mycobacterium tuberculosis |
6.3.1.5 | 8.8 | - |
with substrate glutamine | Mycobacterium tuberculosis |
6.3.5.1 | 8 | - |
recombinant NadE-679, nitrogen donor NH4+ | Mycobacterium tuberculosis |
6.3.5.1 | 8.5 | - |
recombinant NadE-738, nitrogen donor NH4+, recombinant NadE-679, nitrogen donor glutamine | Mycobacterium tuberculosis |
6.3.5.1 | 8.8 | - |
recombinant NadE-738, nitrogen donor glutamine | Mycobacterium tuberculosis |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
6.3.1.5 | ATP | required | Mycobacterium tuberculosis |