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Literature summary extracted from

  • Munawar, N.; Engel, P.C.
    Overexpression in a non-native halophilic host and biotechnological potential of NAD+-dependent glutamate dehydrogenase from Halobacterium salinarum strain NRC-36014 (2012), Extremophiles, 16, 463-476.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.4.1.2 DMSO activates Halobacterium salinarum
1.4.1.2 glycerol activates at 10% Halobacterium salinarum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.1.2 overexpression in halophilic host Haloferax volcanii strain DS70 of soluble, active enzyme with NAD+-GDH activity about 800times higher in the induced sample as compared to controls, expression in inclusion bodies in mesophilic Escherichia coli Halobacterium salinarum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.1.2 0.000011
-
 L-glutamate pH 9.0, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 0.000025
-
 L-glutamate pH 9.0, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 0.00013
-
 NH3 pH 8.5, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 0.00035
-
 NH3 pH 8.5, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 0.0008
-
 2-oxoglutarate pH 8.5, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 0.0034
-
 2-oxoglutarate pH 8.5, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 0.009
-
 NADH pH 8.5, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 0.014
-
 NADH pH 8.5, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 0.112
-
 NAD+ pH 9.0, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 0.118
-
 NAD+ pH 9.0, 40°C, recombinant enzyme Halobacterium salinarum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.4.1.2 betaine high concentrations of salt may be substituted with 30% DMSO or betaine with good stability and activity Halobacterium salinarum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.4.1.2 L-glutamate + H2O + NAD+ Halobacterium salinarum
-
 2-oxoglutarate + NH3 + NADH + H+
-
 r
1.4.1.2 L-glutamate + H2O + NAD+ Halobacterium salinarum NRC-36014
-
 2-oxoglutarate + NH3 + NADH + H+
-
 r

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
1.4.1.2 2-propanol GDHX retains 33% activity in 10% after 72 h, pH 8.5, room temperature. GDHX loses 94% of activity within 3 days in 10% solution Halobacterium salinarum
1.4.1.2 Acetone GDHX retains 38% activity in 10% after 72 h, pH 8.5, room temperature Halobacterium salinarum
1.4.1.2 acetonitrile GDHX retains 30% activity in 10% after 72 h, pH 8.5, room temperature Halobacterium salinarum
1.4.1.2 dimethylformamide 51% activity remaining after 72 h, pH 8.5, room temperature Halobacterium salinarum
1.4.1.2 dimethylformamide GDHX retains 73% activity in 10% DMF after 30 min at pH 8.5, 40°C, and 51% after 72 h, pH 8.5, room temperature Halobacterium salinarum
1.4.1.2 DMSO GDHX retains 100% activity in the aqueous DMSO mixtures, high concentrations of salt may be substituted with 30% DMSO or betaine with good stability and activity Halobacterium salinarum
1.4.1.2 Ethanol GDHX retains 34% activity in 10% after 72 h, pH 8.5, room temperature Halobacterium salinarum
1.4.1.2 Glycerol GDHX retains 73% activity in 10% glycerol after 72 h, pH 8.5, room temperature Halobacterium salinarum
1.4.1.2 Methanol GDHX retains more than 73% activity in 10% after 72 h, pH 8.5, room temperature Halobacterium salinarum

Organism

EC Number Organism UniProt Comment Textmining
1.4.1.2 Halobacterium salinarum P29051
-
-
1.4.1.2 Halobacterium salinarum NRC-36014 P29051
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.1.2 recombinant soluble enzyme 11.3fold from Haloferax volcanii strain DS70, by hydrophobic interaction and anion exchange chromatography, method optimization Halobacterium salinarum

Renatured (Commentary)

EC Number Renatured (Comment) Organism
1.4.1.2 enzyme solubulization from inclusion bodies after expressionin Escherichia coli in 50 mM HEPES-NaOH, pH 7.5, containing 8 M urea and 25 mM DTT, and renaturation of protein using both rapid dilution and gradual dialysis methods Halobacterium salinarum

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.4.1.2 6.52
-
 recombinant enzyme in Haloferax volcanii, pH 8.5, 40°C Halobacterium salinarum
1.4.1.2 35
-
 recombinant enzyme purified from Haloferax volcanii, pH 8.5, 40°C Halobacterium salinarum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.1.2 L-glutamate + H2O + NAD+
-
 Halobacterium salinarum 2-oxoglutarate + NH3 + NADH + H+
-
 r
1.4.1.2 L-glutamate + H2O + NAD+
-
 Halobacterium salinarum NRC-36014 2-oxoglutarate + NH3 + NADH + H+
-
 r

Synonyms

EC Number Synonyms Comment Organism
1.4.1.2 NAD+-dependent GDHX
-
 Halobacterium salinarum
1.4.1.2 NAD+-dependent glutamate dehydrogenase
-
 Halobacterium salinarum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.4.1.2 40
-
 assay at Halobacterium salinarum

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.4.1.2 40 90 temperature profile, overview Halobacterium salinarum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.4.1.2 40 70 recombinant purified enzyme, fully stable at 40°C Halobacterium salinarum
1.4.1.2 80
-
 recombinant purified enzyme, 30 min, 88% remaining Halobacterium salinarum
1.4.1.2 90
-
 recombinant purified enzyme, 30 min, 56% remaining Halobacterium salinarum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4.1.2 15
-
 NAD+ pH 9.0, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 16
-
 NAD+ pH 9.0, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 17
-
 L-glutamate pH 9.0, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 24
-
 L-glutamate pH 9.0, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 39
-
 NADH pH 8.5, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 46
-
 2-oxoglutarate pH 8.5, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 47
-
 NADH pH 8.5, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 56
-
 2-oxoglutarate pH 8.5, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 84
-
 NH3 pH 8.5, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 85
-
 NH3 pH 8.5, 40°C, recombinant enzyme Halobacterium salinarum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.1.2 8.5
-
 amination Halobacterium salinarum
1.4.1.2 9
-
 deamination Halobacterium salinarum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.4.1.2 6.5 10.5 pH profile, overvew Halobacterium salinarum

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.1.2 NAD+
-
 Halobacterium salinarum
1.4.1.2 NADH
-
 Halobacterium salinarum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.4.1.2 0.0018
-
 L-glutamate pH 9.0, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 0.0035
-
 L-glutamate pH 9.0, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 0.72
-
 NAD+ pH 9.0, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 0.75
-
 NAD+ pH 9.0, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 1.7
-
 NADH pH 8.5, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 1.9
-
 NADH pH 8.5, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 16.5
-
 2-oxoglutarate pH 8.5, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 45
-
 NH3 pH 8.5, 40°C, recombinant enzyme Halobacterium salinarum
1.4.1.2 57.5
-
 2-oxoglutarate pH 8.5, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum
1.4.1.2 115
-
 NH3 pH 8.5, 40°C, recombinant enzyme in presence of DMSO Halobacterium salinarum