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Literature summary extracted from

  • Bellinzoni, M.; De Rossi, E.; Branzoni, M.; Milano, A.; Peverali, F.A.; Rizzi, M.; Riccardi, G.
    Heterologous expression, purification, and enzymatic activity of Mycobacterium tuberculosis NAD(+) synthetase (2002), Protein Expr. Purif., 25, 547-557.
    View publication on PubMed

Cloned(Commentary)

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

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.1.5 thrombin enzyme is highly susceptible too proteolytic cleavage Mycobacterium tuberculosis

KM Value [mM]

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

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.1.5 Mg2+ required Mycobacterium tuberculosis

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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
-
?

Organism

EC Number Organism UniProt Comment Textmining
6.3.1.5 Mycobacterium tuberculosis
-
-
-
6.3.5.1 Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Storage Stability

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

Substrates and Products (Substrate)

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+
-
?

Subunits

EC Number Subunits Comment Organism
6.3.5.1 octamer 8 * 67000, SDS-PAGE Mycobacterium tuberculosis

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.3.1.5 37
-
assay at Mycobacterium tuberculosis

pH Optimum

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

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.1.5 ATP required Mycobacterium tuberculosis