EC Number | Application | Comment | Organism |
---|---|---|---|
3.5.1.19 | analysis | nicotinamidases are key analytical biocatalysts for screening modulators in relevant biomedical enzymes, such as sirtuins and poly-ADP-ribosyltransferases | uncultured bacterium |
3.5.1.19 | medicine | the absence of nicotinamidase in humans makes it a promising drug target | unidentified |
EC Number | Cloned (Comment) | Organism |
---|---|---|
3.5.1.B15 | combined whole-cell high-throughput functional screening for identification of nicotinamidases/pyrazinamidases in metagenomic/polygenomic libraries. A fosmid polygenomic expression library obtained from deep-sea mesophilic bacteria is screened, discovering several positive clones with the ammonium ferrous sulfate method. Quantitative rescreening with the SNP method allowing the finding of the first nicotinamidase with balanced catalytic efficiency toward nicotinamidase activity (EC 3.5.1.19) and pyrazinamide (pyrazinamidase activity). Escherichia coli EPI300-T1 is used for the screening of the genomic libraries cloned in pCC1FOS fosmids library. Escherichia coli strain Rosetta2 (DE3) is used as the host for pET46Ek/LIC expression vector. The fosmid library contains inserts of mixed genomic DNA of 194 mesophilic bacteria (MB) isolated from deep-sea hydrothermal vents sampled at various sites in Mid Atlantic Ridge, genotyping, DNA and amino acid sequence determination and analysis | uncultured bacterium |
3.5.1.19 | combined whole-cell high-throughput functional screening for identification of nicotinamidases/pyrazinamidases in metagenomic/polygenomic libraries. A fosmid polygenomic expression library obtained from deep-sea mesophilic bacteria is screened, discovering several positive clones with the ammonium ferrous sulfate method. Quantitative rescreening with the SNP method allowing the finding of the first nicotinamidase with balanced catalytic efficiency toward NAM (nicotinamidase activity) and pyrazinamide (pyrazinamidase activity). Escherichia coli EPI300-T1 is used for the screening of the genomic libraries cloned in pCC1FOS fosmids library. Escherichia coli strain Rosetta2 (DE3) is used as the host for pET46Ek/LIC expression vector. The fosmid library contains inserts of mixed genomic DNA of 194 mesophilic bacteria (MB) isolated from deep-sea hydrothermal vents sampled at various sites in Mid Atlantic Ridge, genotyping, DNA and amino acid sequence determination and analysis | uncultured bacterium |
3.5.1.19 | expression in Escherichia coli Rosetta 2DE3 | unidentified |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
3.5.1.B15 | 5-bromo-nicotinaldehyde | - |
uncultured bacterium | |
3.5.1.B15 | nicotinaldehyde | - |
uncultured bacterium | |
3.5.1.19 | 5-bromo-nicotinaldehyde | - |
uncultured bacterium | |
3.5.1.19 | 5-bromonicotinaldehyde | - |
unidentified | |
3.5.1.19 | nicotinaldehyde | - |
uncultured bacterium | |
3.5.1.19 | nicotinaldehyde | - |
unidentified |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.1.19 | 0.049 | - |
nicotinamide | pH 7.3, temperature not specified in the publication | unidentified | |
3.5.1.19 | 0.103 | - |
Pyrazinamide | pH 7.3, temperature not specified in the publication | unidentified | |
3.5.1.19 | 0.355 | - |
methylnicotinate | pH 7.3, temperature not specified in the publication | unidentified | |
3.5.1.19 | 0.435 | - |
5-Methylnicotinamide | pH 7.3, temperature not specified in the publication | unidentified | |
3.5.1.19 | 1.092 | - |
ethylnicotinate | pH 7.3, temperature not specified in the publication | unidentified |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
3.5.1.19 | Fe2+ | the conserved specific metal ion binding includes one aspartate (D67) and two histidines (H69 and H86) | uncultured bacterium |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
3.5.1.19 | 50300 | - |
gel filtration | unidentified |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.5.1.B15 | pyrazinamide + H2O | uncultured bacterium | - |
pyrazinoic acid + NH3 | - |
r | |
3.5.1.19 | nicotinamide + H2O | unidentified | the enzyme is an essential component of the NAD+ salvage pathway | nicotinate + NH3 | - |
? | |
3.5.1.19 | nicotinamide + H2O | uncultured bacterium | - |
nicotinic acid + NH3 | - |
r |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.5.1.B15 | uncultured bacterium | - |
mesophilic bacteria (MB) isolated from deep-sea hydrothermal vents sampled at various sites in Mid Atlantic Ridge | - |
3.5.1.19 | no activity in Homo sapiens | - |
- |
- |
3.5.1.19 | uncultured bacterium | - |
mesophilic bacteria (MB) isolated from deep-sea hydrothermal vents sampled at various sites in Mid Atlantic Ridge | - |
3.5.1.19 | unidentified | - |
marine metagenome | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.5.1.19 | - |
unidentified |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.5.1.B15 | pyrazinamide + H2O | - |
uncultured bacterium | pyrazinoic acid + NH3 | - |
r | |
3.5.1.19 | 5-methylnicotinamide + H2O | - |
unidentified | 5-methylnicotinate + NH3 | - |
? | |
3.5.1.19 | 5-methylnicotinamide + H2O | - |
uncultured bacterium | 5-methylnicotinic acid + NH3 | - |
r | |
3.5.1.19 | ethylnicotinate + H2O | - |
unidentified | nicotinate + ethanol | - |
? | |
3.5.1.19 | ethylnicotinate + H2O | - |
uncultured bacterium | nicotinic acid + ethanol | - |
r | |
3.5.1.19 | methylnicotinate + H2O | - |
unidentified | nicotinate + methanol | - |
? | |
3.5.1.19 | nicotinamide + H2O | - |
unidentified | nicotinate + NH3 | - |
? | |
3.5.1.19 | nicotinamide + H2O | the enzyme is an essential component of the NAD+ salvage pathway | unidentified | nicotinate + NH3 | - |
? | |
3.5.1.19 | nicotinamide + H2O | - |
uncultured bacterium | nicotinic acid + NH3 | - |
r | |
3.5.1.19 | pyrazinamide + H2O | - |
unidentified | pyrazinoic acid + NH3 | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
3.5.1.19 | dimer | 2 * 23000, SDS-PAGE | unidentified |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.5.1.B15 | More | cf. EC 3.5.1.19 | uncultured bacterium |
3.5.1.B15 | PolyNic | - |
uncultured bacterium |
3.5.1.19 | polygenomic nicotinamidase | - |
unidentified |
3.5.1.19 | PolyNic | - |
unidentified |
3.5.1.19 | PolyNic | - |
uncultured bacterium |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.5.1.B15 | 37 | - |
assay at | uncultured bacterium |
3.5.1.19 | 50 | - |
- |
unidentified |
3.5.1.19 | 50 | - |
at 37°C | uncultured bacterium |
EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.5.1.19 | 10 | 90 | activity range, profile overview | uncultured bacterium |
3.5.1.19 | 37 | 55 | 37°C: about 50% of maximal activity, 55°C: about 60% of maximal activity | unidentified |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.1.19 | 1.1 | - |
ethylnicotinate | pH 7.3, temperature not specified in the publication | unidentified | |
3.5.1.19 | 5 | - |
nicotinamide | pH 7.3, temperature not specified in the publication | unidentified | |
3.5.1.19 | 7 | - |
methylnicotinate | pH 7.3, temperature not specified in the publication | unidentified | |
3.5.1.19 | 7.2 | - |
Pyrazinamide | pH 7.3, temperature not specified in the publication | unidentified | |
3.5.1.19 | 8.2 | - |
5-Methylnicotinamide | pH 7.3, temperature not specified in the publication | unidentified |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.5.1.B15 | 7.3 | - |
assay at | uncultured bacterium |
3.5.1.19 | 10 | - |
- |
unidentified |
3.5.1.19 | 10 | - |
at pH 8.0 | uncultured bacterium |
EC Number | pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|---|
3.5.1.19 | 4 | 11 | activity range, profile overview | uncultured bacterium |
3.5.1.19 | 6 | 10.5 | pH 6.0: about 40% of maximal activity, pH 10.5: about 45% of maximal activity | unidentified |
EC Number | Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.1.B15 | 0.000015 | - |
nicotinaldehyde | pH 7.3, 37°C | uncultured bacterium | |
3.5.1.B15 | 0.00007 | - |
5-bromo-nicotinaldehyde | pH 7.3, 37°C | uncultured bacterium | |
3.5.1.19 | 0.000015 | - |
nicotinaldehyde | pH 7.3, temperature not specified in the publication, substrate: pyrazinamide | unidentified | |
3.5.1.19 | 0.00007 | - |
5-bromonicotinaldehyde | pH 7.3, temperature not specified in the publication, substrate: pyrazinamide | unidentified |
EC Number | General Information | Comment | Organism |
---|---|---|---|
3.5.1.B15 | additional information | combined whole-cell high-throughput functional screening for identification of nicotinamidases/pyrazinamidases in metagenomic/polygenomic libraries, screening of mesophilic marine bacteria (MB) polygenomic library. Development of two whole-cell methods using the chemical property of one of the products formed in the enzymatic reaction (pyrazinoic or NA) to form colored complexes with stable iron salts, such as ammonium ferrous sulfate or sodium nitroprusside (SNP), optimization of the assay. A fosmid polygenomic expression library obtained from deep-sea mesophilic bacteria is screened, discovering several positive clones with the ammonium ferrous sulfate method. Quantitative rescreening with the SNP method allowing the finding of the first nicotinamidase with balanced catalytic efficiency toward nicotinamidase activity (EC 3.5.1.19) and pyrazinamide (pyrazinamidase activity) | uncultured bacterium |
3.5.1.19 | additional information | combined whole-cell high-throughput functional screening for identification of nicotinamidases/pyrazinamidases in metagenomic/polygenomic libraries, screening of mesophilic marine bacteria (MB) polygenomic library. Development of two whole-cell methods using the chemical property of one of the products formed in the enzymatic reaction (pyrazinoic or NA) to form colored complexes with stable iron salts, such as ammonium ferrous sulfate or sodium nitroprusside (SNP), optimization of the assay. A fosmid polygenomic expression library obtained from deep-sea mesophilic bacteria is screened, discovering several positive clones with the ammonium ferrous sulfate method. Quantitative rescreening with the SNP method allowing the finding of the first nicotinamidase with balanced catalytic efficiency toward NAM (nicotinamidase activity) and pyrazinamide (pyrazinamidase activity). PolyNic contains all the conserved catalytic triad residues of the cysteine-hydrolases family:a catalytic cysteine at position 150, an aspartate at position 25, and a lysine at position 111. Other residues important for substrate and metal binding are F30, L36, W83, A112, Y120, and V149 | uncultured bacterium |
3.5.1.19 | physiological function | the enzyme is an essential component of the NAD+ salvage pathway | unidentified |
3.5.1.19 | physiological function | nicotinamidases catalyze the hydrolysis of the amide bond in nicotinamide (NAM) to produce ammonia and nicotinic acid (NA). These enzymes are an essential component of the NAD+ salvage pathway and are implicated in the viability of several pathogenic organisms | uncultured bacterium |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.1.19 | 1 | - |
ethylnicotinate | pH 7.3, temperature not specified in the publication | unidentified | |
3.5.1.19 | 18.9 | - |
5-Methylnicotinamide | pH 7.3, temperature not specified in the publication | unidentified | |
3.5.1.19 | 19.7 | - |
methylnicotinate | pH 7.3, temperature not specified in the publication | unidentified | |
3.5.1.19 | 69.9 | - |
Pyrazinamide | pH 7.3, temperature not specified in the publication | unidentified | |
3.5.1.19 | 102 | - |
nicotinamide | pH 7.3, temperature not specified in the publication | unidentified |