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Literature summary for 6.3.1.5 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)

Cloned (Comment) Organism
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

Inhibitors

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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00052
-
deamido-NAD+ pH 8.5, 37°C, recombinant truncated enzyme Mycobacterium tuberculosis
0.00079
-
ATP pH 8.5, 37°C, recombinant truncated enzyme Mycobacterium tuberculosis
0.0011
-
glutamine pH 8.5, 37°C, recombinant full length enzyme Mycobacterium tuberculosis
0.0013
-
ATP pH 8.5, 37°C, recombinant full length enzyme Mycobacterium tuberculosis
0.0016
-
glutamine pH 8.5, 37°C, recombinant truncated enzyme Mycobacterium tuberculosis
0.0029
-
deamido-NAD+ pH 8.5, 37°C, recombinant full length enzyme Mycobacterium tuberculosis
0.025
-
NH4+ pH 8.5, 37°C, recombinant truncated enzyme Mycobacterium tuberculosis
0.027
-
NH4+ pH 8.5, 37°C, recombinant full length enzyme Mycobacterium tuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + deamido-NAD+ + NH4+ Mycobacterium tuberculosis
-
AMP + diphosphate + NAD+ + H+
-
?

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.009
-
partially purified recombinant His-tagged enzyme expressed in insect cells, substrate NH4+ Mycobacterium tuberculosis
0.033
-
purified recombinant HIs-tagged enzyme expressed in Escherichia coli, substrate glutamine Mycobacterium tuberculosis
0.1
-
purified recombinant His-tagged enzyme expressed in Escherichia coli, substrate NH4+ Mycobacterium tuberculosis
0.67
-
purified recombinant His-tagged truncated enzyme expressed in Escherichia coli, substrate NH4+ Mycobacterium tuberculosis

Storage Stability

Storage Stability Organism
-20°C, recombinant truncated enzyme, overnight, loss of 59% activity Mycobacterium tuberculosis
-70°C, recombinant truncated enzyme, overnight, loss of 30% activity Mycobacterium tuberculosis
-80 - 4°C, recombinant full length enzyme, several months Mycobacterium tuberculosis
25°C, recombinant truncated enzyme, stable Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + deamido-NAD+ + glutamine
-
Mycobacterium tuberculosis AMP + diphosphate + NAD+ + glutamate
-
?
ATP + deamido-NAD+ + NH4+
-
Mycobacterium tuberculosis AMP + diphosphate + NAD+ + H+
-
?

Synonyms

Synonyms Comment Organism
NAD synthase
-
Mycobacterium tuberculosis
NAD synthetase
-
Mycobacterium tuberculosis
Nicotinamide adenine dinucleotide synthetase
-
Mycobacterium tuberculosis

Temperature Optimum [°C]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
with substrate NH4+ Mycobacterium tuberculosis
8.8
-
with substrate glutamine Mycobacterium tuberculosis

Cofactor

Cofactor Comment Organism Structure
ATP required Mycobacterium tuberculosis