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Biochemical and mutational analysis of a novel nicotinamidase from Oceanobacillus iheyensis HTE831

Sanchez-Carron, G.; Garcia-Garcia, M.I.; Zapata-Perez, R.; Takami, H.; Garcia-Carmona, F.; Sanchez-Ferrer, A.; PLoS ONE 8, e56727 (2013)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
3.5.1.19
phylogenetic analysis, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta 2 (DE3)
Oceanobacillus iheyensis
Engineering
EC Number
Amino acid exchange
Commentary
Organism
3.5.1.19
C133A
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
E65H
site-directed mutagenesis, the mutation highly reduces the activity with pyrazinamide compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
F68W
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
F68W/C133A
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
K104A
site-directed mutagenesis, the mutant shows 98.55% reduced activity compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
additional information
mutation of residues Cys133 and Phe68 is required for increasing pyrazinamidase activity 2.9 and 2.5fold, respectively. In addition, the change in the fourth residue involved in the ion metal binding (Glu65) is detrimental to pyrazinamidase activity decreasing it 6fold
Oceanobacillus iheyensis
3.5.1.19
Q96A
site-directed mutagenesis, the mutant shows similar activity with nicotinamide and increased activity with 5-methyl nicotinamide compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
Q96K
site-directed mutagenesis, the mutant shows 62.7% reduced activity compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
T12Q
site-directed mutagenesis, the mutant shows 98.42% reduced activity compared to the wild-type enzyme
Oceanobacillus iheyensis
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.1.19
5-bromonicotinaldehyde
competitive inhibition
Oceanobacillus iheyensis
3.5.1.19
nicotinaldehyde
competitive inhibition
Oceanobacillus iheyensis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.1.19
additional information
-
additional information
Michaelis-Menten kinetics
Oceanobacillus iheyensis
3.5.1.19
0.13
-
nicotinamide
recombinant mutant E65H, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.17
-
nicotinamide
recombinant mutant F68W, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.19
-
nicotinamide
recombinant mutant F68W/C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.26
-
nicotinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.34
-
ethyl nicotinate
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.36
-
nicotinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.36
-
Pyrazinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.4
-
nicotinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.44
-
methyl nicotinate
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.5
-
Pyrazinamide
recombinant mutant F68W, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.55
-
5-methyl nicotinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.56
-
methyl nicotinate
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.57
-
nicotinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.62
-
5-methyl nicotinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.64
-
5-methyl nicotinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.68
-
5-methyl nicotinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.81
-
Pyrazinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.82
-
Pyrazinamide
recombinant mutant E65H, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.83
-
Pyrazinamide
recombinant mutant F68W/C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.89
-
methyl nicotinate
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.02
-
ethyl nicotinate
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.03
-
methyl nicotinate
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.11
-
ethyl nicotinate
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.15
-
Pyrazinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.2
-
Pyrazinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.43
-
ethyl nicotinate
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.5.1.19
additional information
the enzyme contains 0.235 mM of Zn2+ in 0.242 mM protein, while it only contains 0.0032 mM of Fe2+ and 0.00077 mM Mn2+
Oceanobacillus iheyensis
3.5.1.19
Zn2+
presence of Zn2+ in the active center of the enzyme in a molecular ratio of 1:1 of protein to metal ion
Oceanobacillus iheyensis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.1.19
21100
-
2 * 21100, mass spectrometry
Oceanobacillus iheyensis
3.5.1.19
42900
-
gel filtration
Oceanobacillus iheyensis
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
Oceanobacillus iheyensis
-
nicotinate + NH3
-
-
?
3.5.1.19
nicotinamide + H2O
Oceanobacillus iheyensis HTE831
-
nicotinate + NH3
-
-
?
3.5.1.19
pyrazinamide + H2O
Oceanobacillus iheyensis
-
pyrazinoic acid + NH3
-
-
?
3.5.1.19
pyrazinamide + H2O
Oceanobacillus iheyensis HTE831
-
pyrazinoic acid + NH3
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.5.1.19
Oceanobacillus iheyensis
Q8ESQ6
deep-sea extremely halotolerant and alkaliphilic organism
-
3.5.1.19
Oceanobacillus iheyensis HTE831
Q8ESQ6
deep-sea extremely halotolerant and alkaliphilic organism
-
Purification (Commentary)
EC Number
Commentary
Organism
3.5.1.19
recombinant His-tagged wild-type and mutant enzymes methylnicotinamide from Escherichia coli strain Rosetta 2 (DE3) by ultrafiltration and nickel affinity chromatography
Oceanobacillus iheyensis
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.5.1.19
0.06
-
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate ethyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
0.13
-
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate ethyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
0.22
-
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate ethyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
0.31
-
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate methyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
0.34
-
purified recombinant mutant C133A, pH 7.3, 37°C, substrate ethyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
0.36
-
purified recombinant mutant K104A, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
0.37
-
purified recombinant mutant T12Q, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
0.45
-
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate methyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
0.46
-
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
0.7
-
purified recombinant mutant E65H, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
1.16
-
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate methyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
1.4
-
purified recombinant mutant C133A, pH 7.3, 37°C, substrate methyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
1.8
-
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
3.62
-
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
3.96
-
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate 5-methyl nicotinamide
Oceanobacillus iheyensis
3.5.1.19
6.39
-
purified recombinant mutant C133A, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
7.43
-
purified recombinant mutant F68W/C133A, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
7.5
-
purified recombinant mutant F68W, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
8.7
-
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
9.01
-
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate 5-methyl nicotinamide
Oceanobacillus iheyensis
3.5.1.19
11.74
-
purified recombinant mutant C133A, pH 7.3, 37°C, substrate 5-methyl nicotinamide
Oceanobacillus iheyensis
3.5.1.19
13.5
-
purified recombinant mutant E65H, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
17.5
-
purified recombinant mutant C133A, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
19.97
-
purified recombinant mutant F68W, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
22.9
-
purified recombinant mutant F68W/C133A, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
23.3
25
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
24.8
-
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
28
-
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate 5-methyl nicotinamide
Oceanobacillus iheyensis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.1.19
5-methyl nicotinamide + H2O
-
735084
Oceanobacillus iheyensis
5-methyl nicotinate + NH3
-
-
-
?
3.5.1.19
5-methyl nicotinamide + H2O
-
735084
Oceanobacillus iheyensis HTE831
5-methyl nicotinate + NH3
-
-
-
?
3.5.1.19
ethyl nicotinate + H2O
-
735084
Oceanobacillus iheyensis
nicotinate + ethanol
-
-
-
?
3.5.1.19
methyl nicotinate + H2O
-
735084
Oceanobacillus iheyensis
nicotinate + methanol
-
-
-
?
3.5.1.19
nicotinamide + H2O
-
735084
Oceanobacillus iheyensis
nicotinate + NH3
-
-
-
?
3.5.1.19
nicotinamide + H2O
preferred substrate
735084
Oceanobacillus iheyensis
nicotinate + NH3
-
-
-
?
3.5.1.19
nicotinamide + H2O
-
735084
Oceanobacillus iheyensis HTE831
nicotinate + NH3
-
-
-
?
3.5.1.19
nicotinamide + H2O
preferred substrate
735084
Oceanobacillus iheyensis HTE831
nicotinate + NH3
-
-
-
?
3.5.1.19
pyrazinamide + H2O
-
735084
Oceanobacillus iheyensis
pyrazinoic acid + NH3
-
-
-
?
3.5.1.19
pyrazinamide + H2O
residues Cys133 and Phe68 are critical for the pyraminidase activity
735084
Oceanobacillus iheyensis
pyrazinoic acid + NH3
-
-
-
?
3.5.1.19
pyrazinamide + H2O
-
735084
Oceanobacillus iheyensis HTE831
pyrazinoic acid + NH3
-
-
-
?
3.5.1.19
pyrazinamide + H2O
residues Cys133 and Phe68 are critical for the pyraminidase activity
735084
Oceanobacillus iheyensis HTE831
pyrazinoic acid + NH3
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
3.5.1.19
dimer
2 * 21100, mass spectrometry
Oceanobacillus iheyensis
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.5.1.19
45
-
-
Oceanobacillus iheyensis
Temperature Range [°C]
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
3.5.1.19
25
55
activity range, below 25°C or above 55°C the enzyme loses most of its activity
Oceanobacillus iheyensis
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.5.1.19
4
20
purified recombinant wild-type enzyme, 20 h, stable
Oceanobacillus iheyensis
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.5.1.19
0.05
-
ethyl nicotinate
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.1
-
ethyl nicotinate
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.14
-
ethyl nicotinate
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.17
-
methyl nicotinate
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.19
-
ethyl nicotinate
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.32
-
methyl nicotinate
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.43
-
Pyrazinamide
recombinant mutant E65H, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.45
-
Pyrazinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.6
-
methyl nicotinate
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.02
-
methyl nicotinate
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.61
-
Pyrazinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
2.16
-
5-methyl nicotinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
2.37
-
nicotinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
2.6
-
Pyrazinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
3.31
-
Pyrazinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
3.94
-
Pyrazinamide
recombinant mutant F68W, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
5.1
-
5-methyl nicotinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
5.21
-
Pyrazinamide
recombinant mutant F68W/C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
5.63
-
nicotinamide
recombinant mutant E65H, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
7.09
-
5-methyl nicotinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
8.79
-
nicotinamide
recombinant mutant F68W, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
9.82
-
nicotinamide
recombinant mutant F68W/C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
11.65
-
nicotinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
13.6
-
nicotinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
15.9
-
5-methyl nicotinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
16.05
-
nicotinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.5.1.19
6
6.5
-
Oceanobacillus iheyensis
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.5.1.19
5
8
activity range, marked decrease in activity below pH 5.0 and above pH 8.0
Oceanobacillus iheyensis
pH Stability
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
3.5.1.19
6
7.3
purified recombinant wild-type enzyme, very stable at pH 6.0 and pH 7.3, where it maintains 40% and 30% residual activity, respectively, after 20 hours of incubation
Oceanobacillus iheyensis
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
3.5.1.19
0.0035
-
nicotinaldehyde
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.0044
-
5-bromonicotinaldehyde
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.1.19
phylogenetic analysis, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta 2 (DE3)
Oceanobacillus iheyensis
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
3.5.1.19
C133A
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
E65H
site-directed mutagenesis, the mutation highly reduces the activity with pyrazinamide compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
F68W
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
F68W/C133A
site-directed mutagenesis, the mutation affectes the active site cavity, and enhances the binding of pyrazinamide compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
K104A
site-directed mutagenesis, the mutant shows 98.55% reduced activity compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
additional information
mutation of residues Cys133 and Phe68 is required for increasing pyrazinamidase activity 2.9 and 2.5fold, respectively. In addition, the change in the fourth residue involved in the ion metal binding (Glu65) is detrimental to pyrazinamidase activity decreasing it 6fold
Oceanobacillus iheyensis
3.5.1.19
Q96A
site-directed mutagenesis, the mutant shows similar activity with nicotinamide and increased activity with 5-methyl nicotinamide compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
Q96K
site-directed mutagenesis, the mutant shows 62.7% reduced activity compared to the wild-type enzyme
Oceanobacillus iheyensis
3.5.1.19
T12Q
site-directed mutagenesis, the mutant shows 98.42% reduced activity compared to the wild-type enzyme
Oceanobacillus iheyensis
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.1.19
5-bromonicotinaldehyde
competitive inhibition
Oceanobacillus iheyensis
3.5.1.19
nicotinaldehyde
competitive inhibition
Oceanobacillus iheyensis
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
3.5.1.19
0.0035
-
nicotinaldehyde
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.0044
-
5-bromonicotinaldehyde
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.1.19
additional information
-
additional information
Michaelis-Menten kinetics
Oceanobacillus iheyensis
3.5.1.19
0.13
-
nicotinamide
recombinant mutant E65H, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.17
-
nicotinamide
recombinant mutant F68W, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.19
-
nicotinamide
recombinant mutant F68W/C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.26
-
nicotinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.34
-
ethyl nicotinate
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.36
-
nicotinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.36
-
Pyrazinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.4
-
nicotinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.44
-
methyl nicotinate
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.5
-
Pyrazinamide
recombinant mutant F68W, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.55
-
5-methyl nicotinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.56
-
methyl nicotinate
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.57
-
nicotinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.62
-
5-methyl nicotinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.64
-
5-methyl nicotinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.68
-
5-methyl nicotinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.81
-
Pyrazinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.82
-
Pyrazinamide
recombinant mutant E65H, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.83
-
Pyrazinamide
recombinant mutant F68W/C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.89
-
methyl nicotinate
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.02
-
ethyl nicotinate
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.03
-
methyl nicotinate
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.11
-
ethyl nicotinate
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.15
-
Pyrazinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.2
-
Pyrazinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.43
-
ethyl nicotinate
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.5.1.19
additional information
the enzyme contains 0.235 mM of Zn2+ in 0.242 mM protein, while it only contains 0.0032 mM of Fe2+ and 0.00077 mM Mn2+
Oceanobacillus iheyensis
3.5.1.19
Zn2+
presence of Zn2+ in the active center of the enzyme in a molecular ratio of 1:1 of protein to metal ion
Oceanobacillus iheyensis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.1.19
21100
-
2 * 21100, mass spectrometry
Oceanobacillus iheyensis
3.5.1.19
42900
-
gel filtration
Oceanobacillus iheyensis
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
Oceanobacillus iheyensis
-
nicotinate + NH3
-
-
?
3.5.1.19
nicotinamide + H2O
Oceanobacillus iheyensis HTE831
-
nicotinate + NH3
-
-
?
3.5.1.19
pyrazinamide + H2O
Oceanobacillus iheyensis
-
pyrazinoic acid + NH3
-
-
?
3.5.1.19
pyrazinamide + H2O
Oceanobacillus iheyensis HTE831
-
pyrazinoic acid + NH3
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.1.19
recombinant His-tagged wild-type and mutant enzymes methylnicotinamide from Escherichia coli strain Rosetta 2 (DE3) by ultrafiltration and nickel affinity chromatography
Oceanobacillus iheyensis
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.5.1.19
0.06
-
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate ethyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
0.13
-
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate ethyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
0.22
-
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate ethyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
0.31
-
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate methyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
0.34
-
purified recombinant mutant C133A, pH 7.3, 37°C, substrate ethyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
0.36
-
purified recombinant mutant K104A, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
0.37
-
purified recombinant mutant T12Q, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
0.45
-
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate methyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
0.46
-
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
0.7
-
purified recombinant mutant E65H, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
1.16
-
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate methyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
1.4
-
purified recombinant mutant C133A, pH 7.3, 37°C, substrate methyl nicotinate
Oceanobacillus iheyensis
3.5.1.19
1.8
-
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
3.62
-
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
3.96
-
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate 5-methyl nicotinamide
Oceanobacillus iheyensis
3.5.1.19
6.39
-
purified recombinant mutant C133A, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
7.43
-
purified recombinant mutant F68W/C133A, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
7.5
-
purified recombinant mutant F68W, pH 7.3, 37°C, substrate pyrazinamide
Oceanobacillus iheyensis
3.5.1.19
8.7
-
purified recombinant mutant Q96K, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
9.01
-
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate 5-methyl nicotinamide
Oceanobacillus iheyensis
3.5.1.19
11.74
-
purified recombinant mutant C133A, pH 7.3, 37°C, substrate 5-methyl nicotinamide
Oceanobacillus iheyensis
3.5.1.19
13.5
-
purified recombinant mutant E65H, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
17.5
-
purified recombinant mutant C133A, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
19.97
-
purified recombinant mutant F68W, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
22.9
-
purified recombinant mutant F68W/C133A, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
23.3
25
purified recombinant wild-type enzyme, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
24.8
-
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate nicotinamide
Oceanobacillus iheyensis
3.5.1.19
28
-
purified recombinant mutant Q96A, pH 7.3, 37°C, substrate 5-methyl nicotinamide
Oceanobacillus iheyensis
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
5-methyl nicotinamide + H2O
-
735084
Oceanobacillus iheyensis
5-methyl nicotinate + NH3
-
-
-
?
3.5.1.19
5-methyl nicotinamide + H2O
-
735084
Oceanobacillus iheyensis HTE831
5-methyl nicotinate + NH3
-
-
-
?
3.5.1.19
ethyl nicotinate + H2O
-
735084
Oceanobacillus iheyensis
nicotinate + ethanol
-
-
-
?
3.5.1.19
methyl nicotinate + H2O
-
735084
Oceanobacillus iheyensis
nicotinate + methanol
-
-
-
?
3.5.1.19
nicotinamide + H2O
-
735084
Oceanobacillus iheyensis
nicotinate + NH3
-
-
-
?
3.5.1.19
nicotinamide + H2O
preferred substrate
735084
Oceanobacillus iheyensis
nicotinate + NH3
-
-
-
?
3.5.1.19
nicotinamide + H2O
-
735084
Oceanobacillus iheyensis HTE831
nicotinate + NH3
-
-
-
?
3.5.1.19
nicotinamide + H2O
preferred substrate
735084
Oceanobacillus iheyensis HTE831
nicotinate + NH3
-
-
-
?
3.5.1.19
pyrazinamide + H2O
-
735084
Oceanobacillus iheyensis
pyrazinoic acid + NH3
-
-
-
?
3.5.1.19
pyrazinamide + H2O
residues Cys133 and Phe68 are critical for the pyraminidase activity
735084
Oceanobacillus iheyensis
pyrazinoic acid + NH3
-
-
-
?
3.5.1.19
pyrazinamide + H2O
-
735084
Oceanobacillus iheyensis HTE831
pyrazinoic acid + NH3
-
-
-
?
3.5.1.19
pyrazinamide + H2O
residues Cys133 and Phe68 are critical for the pyraminidase activity
735084
Oceanobacillus iheyensis HTE831
pyrazinoic acid + NH3
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.5.1.19
dimer
2 * 21100, mass spectrometry
Oceanobacillus iheyensis
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.5.1.19
45
-
-
Oceanobacillus iheyensis
Temperature Range [°C] (protein specific)
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
3.5.1.19
25
55
activity range, below 25°C or above 55°C the enzyme loses most of its activity
Oceanobacillus iheyensis
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.5.1.19
4
20
purified recombinant wild-type enzyme, 20 h, stable
Oceanobacillus iheyensis
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
0.05
-
ethyl nicotinate
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.1
-
ethyl nicotinate
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.14
-
ethyl nicotinate
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.17
-
methyl nicotinate
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.19
-
ethyl nicotinate
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.32
-
methyl nicotinate
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.43
-
Pyrazinamide
recombinant mutant E65H, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.45
-
Pyrazinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.6
-
methyl nicotinate
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.02
-
methyl nicotinate
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.61
-
Pyrazinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
2.16
-
5-methyl nicotinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
2.37
-
nicotinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
2.6
-
Pyrazinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
3.31
-
Pyrazinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
3.94
-
Pyrazinamide
recombinant mutant F68W, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
5.1
-
5-methyl nicotinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
5.21
-
Pyrazinamide
recombinant mutant F68W/C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
5.63
-
nicotinamide
recombinant mutant E65H, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
7.09
-
5-methyl nicotinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
8.79
-
nicotinamide
recombinant mutant F68W, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
9.82
-
nicotinamide
recombinant mutant F68W/C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
11.65
-
nicotinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
13.6
-
nicotinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
15.9
-
5-methyl nicotinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
16.05
-
nicotinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.5.1.19
6
6.5
-
Oceanobacillus iheyensis
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.5.1.19
5
8
activity range, marked decrease in activity below pH 5.0 and above pH 8.0
Oceanobacillus iheyensis
pH Stability (protein specific)
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
3.5.1.19
6
7.3
purified recombinant wild-type enzyme, very stable at pH 6.0 and pH 7.3, where it maintains 40% and 30% residual activity, respectively, after 20 hours of incubation
Oceanobacillus iheyensis
General Information
EC Number
General Information
Commentary
Organism
3.5.1.19
metabolism
nicotinamidases catalyze the hydrolysis of nicotinamide to nicotinic acid and ammonia, an important reaction in the NAD+ salvage pathway
Oceanobacillus iheyensis
3.5.1.19
additional information
two for residues involved in the catalysis (K104 and Q96), one for the fourth residue involved in metal binding (E65), one for the residue forming hydrogen bonds between the main and lateral chains (T12), one for the residue involved in the cis-peptide bond and in the formation of the oxyanion hole (C133), and one for the residue forming one of the faces of the active site F68
Oceanobacillus iheyensis
General Information (protein specific)
EC Number
General Information
Commentary
Organism
3.5.1.19
metabolism
nicotinamidases catalyze the hydrolysis of nicotinamide to nicotinic acid and ammonia, an important reaction in the NAD+ salvage pathway
Oceanobacillus iheyensis
3.5.1.19
additional information
two for residues involved in the catalysis (K104 and Q96), one for the fourth residue involved in metal binding (E65), one for the residue forming hydrogen bonds between the main and lateral chains (T12), one for the residue involved in the cis-peptide bond and in the formation of the oxyanion hole (C133), and one for the residue forming one of the faces of the active site F68
Oceanobacillus iheyensis
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
0.05
-
ethyl nicotinate
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.1
-
ethyl nicotinate
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.13
-
ethyl nicotinate
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.31
-
methyl nicotinate
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.38
-
methyl nicotinate
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.39
-
Pyrazinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.41
-
ethyl nicotinate
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.52
-
Pyrazinamide
recombinant mutant E65H, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.14
-
methyl nicotinate
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.17
-
methyl nicotinate
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.34
-
Pyrazinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
3.2
-
Pyrazinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
3.4
-
5-methyl nicotinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
4.15
-
nicotinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
6.28
-
Pyrazinamide
recombinant mutant F68W/C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
7.5
-
5-methyl nicotinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
7.9
-
Pyrazinamide
recombinant mutant F68W, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
9.19
-
Pyrazinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
11.07
-
5-methyl nicotinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
29.04
-
5-methyl nicotinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
34
-
nicotinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
42.65
-
nicotinamide
recombinant mutant E65H, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
43.48
-
nicotinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
44.59
-
nicotinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
52.01
-
nicotinamide
recombinant mutant F68W, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
52.8
-
nicotinamide
recombinant mutant F68W/C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
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
0.05
-
ethyl nicotinate
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.1
-
ethyl nicotinate
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.13
-
ethyl nicotinate
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.31
-
methyl nicotinate
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.38
-
methyl nicotinate
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.39
-
Pyrazinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.41
-
ethyl nicotinate
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
0.52
-
Pyrazinamide
recombinant mutant E65H, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.14
-
methyl nicotinate
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.17
-
methyl nicotinate
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
1.34
-
Pyrazinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
3.2
-
Pyrazinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
3.4
-
5-methyl nicotinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
4.15
-
nicotinamide
recombinant mutant Q96K, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
6.28
-
Pyrazinamide
recombinant mutant F68W/C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
7.5
-
5-methyl nicotinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
7.9
-
Pyrazinamide
recombinant mutant F68W, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
9.19
-
Pyrazinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
11.07
-
5-methyl nicotinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
29.04
-
5-methyl nicotinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
34
-
nicotinamide
recombinant mutant C133A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
42.65
-
nicotinamide
recombinant mutant E65H, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
43.48
-
nicotinamide
recombinant wild-type enzyme, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
44.59
-
nicotinamide
recombinant mutant Q96A, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
52.01
-
nicotinamide
recombinant mutant F68W, pH 7.3, 37°C
Oceanobacillus iheyensis
3.5.1.19
52.8
-
nicotinamide
recombinant mutant F68W/C133A, pH 7.3, 37°C
Oceanobacillus iheyensis