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Literature summary for 6.3.1.11 extracted from

  • Kurihara, S.; Oda, S.; Tsuboi, Y.; Kim, H.G.; Oshida, M.; Kumagai, H.; Suzuki, H.
    gamma-Glutamylputrescine synthetase in the putrescine utilization pathway of Escherichia coli K-12 (2008), J. Biol. Chem., 283, 19981-19990.
    View publication on PubMed
Search for more literature for 6.3.1.11

Activating Compound

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(Commentary)

Cloned (Comment) Organism
gene puuA, expression analysis, expression of untagged enzyme Escherichia coli K-12

Protein Variants

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 [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

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Escherichia coli K-12
-
 gene puuA
-

Purification (Commentary)

Purification (Comment) Organism
recombinant untagged enzyme by ammonium sulfate fractionation and adsorption chromatography Escherichia coli K-12

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
12.7
-
 purified recombinant enzyme Escherichia coli K-12

Substrates and Products (Substrate)

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

Subunits Comment Organism
dodecamer 12 * 53000, recombinant enzyme, SDS-PAGE Escherichia coli K-12

Synonyms

Synonyms Comment Organism
gamma-glutamylputrescine synthetase
-
 Escherichia coli K-12
PuuA
-
 Escherichia coli K-12

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Escherichia coli K-12

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
 Escherichia coli K-12

Cofactor

Cofactor Comment Organism Structure
ATP dependent on Escherichia coli K-12