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Literature summary extracted from

  • Auger, G.; Martin, L.; Bertrand, J.; Ferrari, P.; Fanchon, E.; Vaganay, S.; Petillot, Y.; van Heijenoort, J.; Blanot, D.; Dideberg, O.
    Large-scale preparation, purification, and crystallization of UDP-N-acetylmuramoyl-L-alanine: D-glutamate ligase from Escherichia coli (1998), Protein Expr. Purif., 13, 23-29.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.2.9 expressed in Escherichia coli JM83(pMLD58) Escherichia coli

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
6.3.2.9 hanging-drop vapor diffusion technique Escherichia coli

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.2.9 arsenate increases storage stability Escherichia coli
6.3.2.9 Ca2+ increases storage stability Escherichia coli
6.3.2.9 Cd2+ abolishes the enzyme activity at 5mM Escherichia coli
6.3.2.9 Mg2+ increases storage stability Escherichia coli
6.3.2.9 Mn2+ increases storage stability Escherichia coli
6.3.2.9 phosphate increases storage stability Escherichia coli
6.3.2.9 sulfate increases storage stability Escherichia coli
6.3.2.9 vanadate abolishes the enzyme activity at 5mM Escherichia coli
6.3.2.9 Zn2+ abolishes the enzyme activity at 5mM Escherichia coli

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.3.2.9 46840
-
 mass spectrometry Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.9 Escherichia coli
-
-
-
6.3.2.9 Escherichia coli JM83(pMLD58)
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.2.9 method that includes DEAE-Trisacryl M and hydroxyapatite-Ultrogel chromatography. Large-scale purification using 2-mercaptoethanol as a reducing agent, results in 200 mg of purified enzyme from 54 l of culture, adducts with one or two molecules of 2-mercaptoethanol are formed. Replacement of 2-mercaptoethanol by dithiothreitol gave 162 mg of enzyme from 36 l of culture yielding no clusters with the reducing agent. Escherichia coli

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.3.2.9 1.772
-
 37°C Escherichia coli

Storage Stability

EC Number Storage Stability Organism
6.3.2.9 -20°C, 20 mM potassium phosphate, pH 7.0, 1 mM 2-mercaptoethanol, 0.1 mM MgCl2, several months, no loss of activity at high enzyme concentrations: 1-10 mg/ml Escherichia coli
6.3.2.9 -20°C, 20 mM potassium phosphate, pH 7.0, 1 mM dithiothreitol, 0.1 mM MgCl2, several months, no loss of activity at high enzyme concentrations: 1-10 mg/ml Escherichia coli
6.3.2.9 8°C, 20 mM potassium phosphate, pH 7.0, 1 mM 2-mercaptoethanol, 0.1 mM MgCl2, 1 week, 30% loss of activity Escherichia coli
6.3.2.9 8°C, 20 mM potassium phosphate, pH 7.0, 1 mM 2-mercaptoethanol, 0.1 mM MgCl2, sodium Hepes, 1 week, 35-43% loss of activity Escherichia coli
6.3.2.9 8°C, 20 mM potassium phosphate, pH 7.0, 1 mM dithiothreitol, 0.1 mM MgCl2, 1 week, 30% loss of activity Escherichia coli
6.3.2.9 8°C, 20 mM potassium phosphate, pH 7.0, 1 mM dithiothreitol, 0.1 mM MgCl2, sodium Hepes 1 week, 35-43% loss of activity Escherichia coli
6.3.2.9 8°C, 20 mM potassium phosphate, pH 8.0, 1 mM 2-mercaptoethanol, 0.1 mM MgCl2, sodium Hepes, 1 week, 35-43% loss of activity Escherichia coli
6.3.2.9 8°C, 20 mM potassium phosphate, pH 8.0, 1 mM dithiothreitol, 0.1 mM MgCl2, sodium Hepes 1 week, 35-43% loss of activity Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.3.2.9 additional information
-
 additional information turnover of 6.55 at saturation concentrations of all the substrates: 5 mM ATP, 0.25 mM UDP-N-acetylmuramoyl L-alanine, 0.55 mM D-glutamate Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.2.9 8.8
-
-
 Escherichia coli