Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | The transcription of puuA is induced by putrescine, deletion of gene puuR leads to overexpression of puuA | Escherichia coli K-12 |
Cloned (Comment) | Organism |
---|---|
gene puuA, expression analysis, expression of untagged enzyme | Escherichia coli K-12 |
Protein Variants | Comment | Organism |
---|---|---|
H269N | site-directed mutagenesis, the mutant shows 3% activity compared to the wild-type enzyme, the mutant also shows increased susceptibility to proteolytic degradation | Escherichia coli K-12 |
H269N/R344Q | site-directed mutagenesis, the mutant shows 0.1% activity compared to the wild-type enzyme, the mutant also shows increased susceptibility to proteolytic degradation | Escherichia coli K-12 |
H282N | site-directed mutagenesis, the mutant shows 9% activity compared to the wild-type enzyme | Escherichia coli K-12 |
H282N/R357Q | site-directed mutagenesis, the mutant shows 0.05% activity compared to the wild-type enzyme | Escherichia coli K-12 |
additional information | deletion of the puu gene cluster, puuA deletion mutants cannot grow on minimal medium with putrescine as the sole source of nitrogen, phenotypes, overview | Escherichia coli K-12 |
R357Q | site-directed mutagenesis, the mutant shows 3% activity compared to the wild-type enzyme | Escherichia coli K-12 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.07 | - |
L-glutamate | pH 8.0 37°C, recombinant enzyme | Escherichia coli K-12 | |
2.35 | - |
ATP | pH 8.0 37°C, recombinant enzyme | Escherichia coli K-12 | |
44.6 | - |
putrescine | pH 8.0 37°C, recombinant enzyme | Escherichia coli K-12 |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required, optimal at 25 mM | Escherichia coli K-12 | |
Mn2+ | can substitute Mg2+ | Escherichia coli K-12 |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
53000 | - |
12 * 53000, recombinant enzyme, SDS-PAGE | Escherichia coli K-12 |
630000 | - |
recombinant enzyme, gel filtration | Escherichia coli K-12 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-glutamate + putrescine | Escherichia coli K-12 | the enzyme catalyzes the gamma-glutamylation of putrescine, the first step in a different putrescine utilization pathway involving gamma-glutamylated intermediates, the Puu pathway, in Escherichia coli, overview. The intracellular concentration of putrescine is probably optimized by PuuA transcriptionally and posttranslationally and excess putrescine is converted to a nutrient source by the Puu pathway, regulation, overview | ADP + phosphate + gamma-L-glutamylputrescine | - |
? | |
additional information | Escherichia coli K-12 | intracellular polyamine profile of various strains relevant to puuA, overview | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli K-12 | - |
gene puuA | - |
Purification (Comment) | Organism |
---|---|
recombinant untagged enzyme by ammonium sulfate fractionation and adsorption chromatography | Escherichia coli K-12 |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
12.7 | - |
purified recombinant enzyme | Escherichia coli K-12 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-glutamate + putrescine | the enzyme catalyzes the gamma-glutamylation of putrescine, the first step in a different putrescine utilization pathway involving gamma-glutamylated intermediates, the Puu pathway, in Escherichia coli, overview. The intracellular concentration of putrescine is probably optimized by PuuA transcriptionally and posttranslationally and excess putrescine is converted to a nutrient source by the Puu pathway, regulation, overview | Escherichia coli K-12 | ADP + phosphate + gamma-L-glutamylputrescine | - |
? | |
ATP + L-glutamate + putrescine | PuuA exhibits the highest activity for putrescine, which has four carbon atoms, and activity drops when the methylene chain of diamine is longer or shorter than C4 | Escherichia coli K-12 | ADP + phosphate + gamma-L-glutamylputrescine | - |
? | |
additional information | intracellular polyamine profile of various strains relevant to puuA, overview | Escherichia coli K-12 | ? | - |
? | |
additional information | when any one of the substrates and a cofactor , i.e. putrescine, glutamate, ATP, and the metal ion, are omitted from the reaction mixture, gamma-Glu-Put is not formed | Escherichia coli K-12 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dodecamer | 12 * 53000, recombinant enzyme, SDS-PAGE | Escherichia coli K-12 |
Synonyms | Comment | Organism |
---|---|---|
gamma-glutamylputrescine synthetase | - |
Escherichia coli K-12 |
PuuA | - |
Escherichia coli K-12 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli K-12 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.94 | - |
putrescine | pH 8.0 37°C, recombinant enzyme | Escherichia coli K-12 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Escherichia coli K-12 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | dependent on | Escherichia coli K-12 |