Cloned (Comment) | Organism |
---|---|
gene pncC, sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
E28A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
K61Q | site-directed mutagenesis, inactive mutant | Escherichia coli |
R142A | site-directed mutagenesis, inactive mutant | Escherichia coli |
S103A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
S29A | site-directed mutagenesis, inactive mutant | Escherichia coli |
T31A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Escherichia coli |
Y56A | site-directed mutagenesis, inactive mutant | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | no inhibition by PMSF and desthiobiotin-fluorophosphonate | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Escherichia coli | |
0.007 | - |
beta-nicotinamide D-ribonucleotide | recombinant wild-type enzyme, pH 7.5, 37°C | Escherichia coli | |
0.012 | - |
beta-nicotinamide D-ribonucleotide | recombinant mutant S103A, pH 7.5, 37°C | Escherichia coli | |
0.027 | - |
beta-nicotinamide D-ribonucleotide | recombinant mutant T31A, pH 7.5, 37°C | Escherichia coli | |
0.272 | - |
beta-nicotinamide D-ribonucleotide | recombinant mutant E28A, pH 7.5, 37°C | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
beta-nicotinamide D-ribonucleotide + H2O | Escherichia coli | - |
beta-nicotinate D-ribonucleotide + NH3 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
gene pncC | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nicke affinity chromatography | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
beta-nicotinamide D-ribonucleotide + H2O = beta-nicotinate D-ribonucleotide + NH3 | proposed mechanism of nicotinamide D-ribonucleotide deamidation by the enzyme via three reaction intermediates, which is a Ser/Lys hydrolase employing a Ser/Lys catalytic dyad, active site structure, overview | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
beta-nicotinamide D-ribonucleotide + H2O | - |
Escherichia coli | beta-nicotinate D-ribonucleotide + NH3 | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | enzyme structure comparisons | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
NMN deamidase | - |
Escherichia coli |
PncC | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
purified proteins, wild-type enzyme loses 40% activity after 20 min, 60% after 40 min, and 80% after 60 min, the mutant E28A lose s 50%, 80% and over 90% activity after 30, 40 qnd 60 min, the mutant T31A is inactivated within 20 min | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.1 | - |
beta-nicotinamide D-ribonucleotide | recombinant mutant S103A, pH 7.5, 37°C | Escherichia coli | |
3.2 | - |
beta-nicotinamide D-ribonucleotide | recombinant mutant T31A, pH 7.5, 37°C | Escherichia coli | |
3.9 | - |
beta-nicotinamide D-ribonucleotide | recombinant mutant E28A, pH 7.5, 37°C | Escherichia coli | |
4.1 | - |
beta-nicotinamide D-ribonucleotide | recombinant wild-type enzyme, pH 7.5, 37°C | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the PncC superfamily, active site configuration, structure comparisons, overview | Escherichia coli |
additional information | enzyme surface charge analysis and substrate docking performed on the enzyme structure from Agrobacterium tumefaciens, PDB code 2A9S, overview | Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
10 | - |
beta-nicotinamide D-ribonucleotide | recombinant mutant E28A, pH 7.5, 37°C | Escherichia coli | |
90 | - |
beta-nicotinamide D-ribonucleotide | recombinant mutant S103A, pH 7.5, 37°C | Escherichia coli | |
120 | - |
beta-nicotinamide D-ribonucleotide | recombinant mutant T31A, pH 7.5, 37°C | Escherichia coli | |
590 | - |
beta-nicotinamide D-ribonucleotide | recombinant wild-type enzyme, pH 7.5, 37°C | Escherichia coli |