Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Saccharomyces cerevisiae |
Crystallization (Comment) | Organism |
---|---|
in complex with nicotinaldehyde, hanging drop vapor diffusion method, using 1.6 M NaOAc, 10% (w/v) ethylene glycol, and 0.1 M HEPES (pH 7.4) | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
C167A | the mutant shows most strongly decreased kcat value compared to the wild type enzyme (below the assay detection limit) | Saccharomyces cerevisiae |
D51A | the zinc-binding mutant shows decreased kcat value compared to the wild type enzyme | Saccharomyces cerevisiae |
D51N | the zinc-binding mutant shows decreased kcat value compared to the wild type enzyme | Saccharomyces cerevisiae |
D8A | the mutant shows 770fold decreased kcat value compared to the wild type enzyme | Saccharomyces cerevisiae |
D8E | the mutant shows 100fold decreased kcat value compared to the wild type enzyme | Saccharomyces cerevisiae |
D8N | the mutant shows 1000fold decreased kcat value compared to the wild type enzyme | Saccharomyces cerevisiae |
H53A | the zinc-binding mutant shows decreased kcat value compared to the wild type enzyme | Saccharomyces cerevisiae |
H94A | the zinc-binding mutant shows decreased kcat value compared to the wild type enzyme | Saccharomyces cerevisiae |
K122A | the mutant shows 16fold decreased kcat value compared to the wild type enzyme | Saccharomyces cerevisiae |
K122R | the mutant shows 770fold decreased kcat value compared to the wild type enzyme | Saccharomyces cerevisiae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
3-acetylpyridine | competitive inhibition | Saccharomyces cerevisiae | |
benzaldehyde | competitive inhibition | Saccharomyces cerevisiae | |
additional information | not inhibited by 10 mM EDTA | Saccharomyces cerevisiae | |
nicotinaldehyde | most potent inhibitor | Saccharomyces cerevisiae | |
nicotinic acid | competitive inhibition | Saccharomyces cerevisiae | |
pyrazinoic acid | competitive inhibition | Saccharomyces cerevisiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0018 | - |
nicotinamide | Km below 0.0018 mM, mutant enzyme D51N, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.0024 | - |
nicotinamide | Km below 0.0024 mM, mutant enzyme D51A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.0027 | - |
nicotinamide | Km below 0.0027 mM, mutant enzyme H94A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.0065 | - |
nicotinamide | Km below 0.0065 mM, mutant enzyme K122A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.0065 | - |
nicotinamide | mutant enzyme H53A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.0096 | - |
nicotinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.025 | - |
Benzamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.061 | - |
5-Methylnicotinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.157 | - |
Pyrazinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | contains tightly bound zinc. Addition of 2 mM ZnCl2 fails to increase the activity of the wild type enzyme | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
nicotinamide + H2O | Saccharomyces cerevisiae | best substrate | nicotinate + NH3 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | P53184 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography and Sephacryl S-300 gel filtration | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-methylnicotinamide + H2O | - |
Saccharomyces cerevisiae | 1-methylnicotinate + NH3 | - |
? | |
5-methylnicotinamide + H2O | - |
Saccharomyces cerevisiae | 5-methylnicotinate + NH3 | - |
? | |
benzamide + H2O | - |
Saccharomyces cerevisiae | benzoate + NH3 | - |
? | |
additional information | no activity with nicotinamide mononucleotide and NAD+ | Saccharomyces cerevisiae | ? | - |
? | |
nicotinamide + H2O | best substrate | Saccharomyces cerevisiae | nicotinate + NH3 | - |
? | |
pyrazinamide + H2O | - |
Saccharomyces cerevisiae | pyrazine-2-carboxylic acid + NH3 | - |
? | |
thionicotinamide + H2O | - |
Saccharomyces cerevisiae | pyridine-3-carbothioic O-acid | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Pnc1 | - |
Saccharomyces cerevisiae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0005 | - |
nicotinamide | kcat below 0.0005 s-1, mutant enzyme C167A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.0006 | - |
nicotinamide | mutant enzyme D8E, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.0009 | - |
nicotinamide | mutant enzyme D8A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.0009 | - |
nicotinamide | mutant enzyme D8N, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.0009 | - |
nicotinamide | mutant enzyme K122R, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.007 | - |
nicotinamide | mutant enzyme D51A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.0088 | - |
Benzamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.014 | - |
nicotinamide | mutant enzyme H53A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.02 | - |
nicotinamide | mutant enzyme H94A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.044 | - |
nicotinamide | mutant enzyme K122A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.064 | - |
nicotinamide | mutant enzyme D51N, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.69 | - |
nicotinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
1.75 | - |
5-Methylnicotinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
2.56 | - |
Pyrazinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | 8 | - |
Saccharomyces cerevisiae |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00094 | - |
nicotinaldehyde | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.0206 | - |
benzaldehyde | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.12 | - |
nicotinic acid | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.316 | - |
3-acetylpyridine | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
6.7 | - |
pyrazinoic acid | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0086 | - |
1-methylnicotinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.0096 | - |
nicotinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.025 | - |
Benzamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.05 | - |
Thionicotinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.061 | - |
5-Methylnicotinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.157 | - |
Pyrazinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
0.35 | - |
Benzamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
2.2 | - |
nicotinamide | mutant enzyme H53A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
6.7 | - |
nicotinamide | kcat/Km above 6.7 (1/sec*mM), mutant enzyme K122A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
7.4 | - |
nicotinamide | kcat/Km above 7.4 (1/sec*mM), mutant enzyme H94A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
8.3 | - |
nicotinamide | kcat/Km above 8.3 (1/sec*mM), mutant enzyme D51A, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
16 | - |
Pyrazinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
29 | - |
5-Methylnicotinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
35 | - |
nicotinamide | kcat/Km above 35 (1/sec*mM), mutant enzyme D51N, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae | |
72 | - |
nicotinamide | wild type enzyme, in 50 mM sodium phosphate (pH 7.5), at 25°C | Saccharomyces cerevisiae |