BRENDA - Enzyme Database show

Kinetics and inhibition of nicotinamidase from Mycobacterium tuberculosis

Seiner, D.R.; Hegde, S.S.; Blanchard, J.S.; Biochemistry 49, 9613-9619 (2010)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
3.5.1.19
expressed in Escherichia coli BL21(DE3) cells
Mycobacterium tuberculosis
Engineering
EC Number
Amino acid exchange
Commentary
Organism
3.5.1.19
H57D
the mutant has reduced Mn2+ content and shows a 6fold and 38fold decrease in kcat value for nicotinamide and pyrazinamide, respectively
Mycobacterium tuberculosis
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.1.19
3-pyridine carboxaldehyde
competitive inhibitor
Mycobacterium tuberculosis
3.5.1.19
iodoacetamide
reversible, competitive inhibitor
Mycobacterium tuberculosis
3.5.1.19
pyrazinecarbonitrile
irreversible inactivator
Mycobacterium tuberculosis
3.5.1.19
pyridine-3-carbonitrile
reversible, competitive inhibitor
Mycobacterium tuberculosis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.1.19
0.013
-
nicotinamide
mutant enzyme H57D, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
0.014
-
nicotinamide
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
0.016
-
Pyrazinamide
mutant enzyme H57D, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
0.3
-
Pyrazinamide
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.5.1.19
Fe2+
the enzyme contains 0.47 M of Fe2+ per mol of enzyme
Mycobacterium tuberculosis
3.5.1.19
Mn2+
the enzyme contains 0.44 M Mn2+ per mol of enzyme
Mycobacterium tuberculosis
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.5.1.19
nicotinamide + H2O
Mycobacterium tuberculosis
-
nicotinate + NH3
-
-
?
3.5.1.19
nicotinamide + H2O
Mycobacterium tuberculosis H37Rv
-
nicotinate + NH3
-
-
?
3.5.1.19
pyrazinamide + H2O
Mycobacterium tuberculosis
-
pyrazine-2-carboxylic acid + NH3
-
-
?
3.5.1.19
pyrazinamide + H2O
Mycobacterium tuberculosis H37Rv
-
pyrazine-2-carboxylic acid + NH3
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.5.1.19
Mycobacterium tuberculosis
-
-
-
3.5.1.19
Mycobacterium tuberculosis H37Rv
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
3.5.1.19
Ni-NTA column chromatography
Mycobacterium tuberculosis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.1.19
nicotinamide + H2O
-
718885
Mycobacterium tuberculosis
nicotinate + NH3
-
-
-
?
3.5.1.19
nicotinamide + H2O
-
718885
Mycobacterium tuberculosis H37Rv
nicotinate + NH3
-
-
-
?
3.5.1.19
pyrazinamide + H2O
-
718885
Mycobacterium tuberculosis
pyrazine-2-carboxylic acid + NH3
-
-
-
?
3.5.1.19
pyrazinamide + H2O
-
718885
Mycobacterium tuberculosis H37Rv
pyrazine-2-carboxylic acid + NH3
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.5.1.19
0.1
-
Pyrazinamide
mutant enzyme H57D, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
0.5
-
nicotinamide
mutant enzyme H57D, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
3.1
-
nicotinamide
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
3.8
-
Pyrazinamide
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.5.1.19
8
-
around pH 8.0
Mycobacterium tuberculosis
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
3.5.1.19
0.00029
-
3-pyridine carboxaldehyde
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
0.1
-
pyridine-3-carbonitrile
Ki about 0.1 mM, wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.1.19
expressed in Escherichia coli BL21(DE3) cells
Mycobacterium tuberculosis
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
3.5.1.19
H57D
the mutant has reduced Mn2+ content and shows a 6fold and 38fold decrease in kcat value for nicotinamide and pyrazinamide, respectively
Mycobacterium tuberculosis
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.1.19
3-pyridine carboxaldehyde
competitive inhibitor
Mycobacterium tuberculosis
3.5.1.19
iodoacetamide
reversible, competitive inhibitor
Mycobacterium tuberculosis
3.5.1.19
pyrazinecarbonitrile
irreversible inactivator
Mycobacterium tuberculosis
3.5.1.19
pyridine-3-carbonitrile
reversible, competitive inhibitor
Mycobacterium tuberculosis
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
3.5.1.19
0.00029
-
3-pyridine carboxaldehyde
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
0.1
-
pyridine-3-carbonitrile
Ki about 0.1 mM, wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.1.19
0.013
-
nicotinamide
mutant enzyme H57D, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
0.014
-
nicotinamide
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
0.016
-
Pyrazinamide
mutant enzyme H57D, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
0.3
-
Pyrazinamide
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.5.1.19
Fe2+
the enzyme contains 0.47 M of Fe2+ per mol of enzyme
Mycobacterium tuberculosis
3.5.1.19
Mn2+
the enzyme contains 0.44 M Mn2+ per mol of enzyme
Mycobacterium tuberculosis
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.5.1.19
nicotinamide + H2O
Mycobacterium tuberculosis
-
nicotinate + NH3
-
-
?
3.5.1.19
nicotinamide + H2O
Mycobacterium tuberculosis H37Rv
-
nicotinate + NH3
-
-
?
3.5.1.19
pyrazinamide + H2O
Mycobacterium tuberculosis
-
pyrazine-2-carboxylic acid + NH3
-
-
?
3.5.1.19
pyrazinamide + H2O
Mycobacterium tuberculosis H37Rv
-
pyrazine-2-carboxylic acid + NH3
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.1.19
Ni-NTA column chromatography
Mycobacterium tuberculosis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.1.19
nicotinamide + H2O
-
718885
Mycobacterium tuberculosis
nicotinate + NH3
-
-
-
?
3.5.1.19
nicotinamide + H2O
-
718885
Mycobacterium tuberculosis H37Rv
nicotinate + NH3
-
-
-
?
3.5.1.19
pyrazinamide + H2O
-
718885
Mycobacterium tuberculosis
pyrazine-2-carboxylic acid + NH3
-
-
-
?
3.5.1.19
pyrazinamide + H2O
-
718885
Mycobacterium tuberculosis H37Rv
pyrazine-2-carboxylic acid + NH3
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.5.1.19
0.1
-
Pyrazinamide
mutant enzyme H57D, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
0.5
-
nicotinamide
mutant enzyme H57D, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
3.1
-
nicotinamide
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
3.8
-
Pyrazinamide
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.5.1.19
8
-
around pH 8.0
Mycobacterium tuberculosis
General Information
EC Number
General Information
Commentary
Organism
3.5.1.19
metabolism
the enzyme is involved in the NAD+ salvage pathway
Mycobacterium tuberculosis
General Information (protein specific)
EC Number
General Information
Commentary
Organism
3.5.1.19
metabolism
the enzyme is involved in the NAD+ salvage pathway
Mycobacterium tuberculosis
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
3.5.1.19
6.2
-
Pyrazinamide
mutant enzyme H57D, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
13
-
Pyrazinamide
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
37
-
nicotinamide
mutant enzyme H57D, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
220
-
nicotinamide
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
3.5.1.19
6.2
-
Pyrazinamide
mutant enzyme H57D, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
13
-
Pyrazinamide
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
37
-
nicotinamide
mutant enzyme H57D, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis
3.5.1.19
220
-
nicotinamide
wild type enzyme, in 100 mM HEPES at pH 8.0 and 25°C
Mycobacterium tuberculosis