Activating Compound | Comment | Organism | Structure |
---|---|---|---|
DMSO | activates | Halobacterium salinarum | |
glycerol | activates at 10% | Halobacterium salinarum |
Cloned (Comment) | Organism |
---|---|
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] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000011 | - |
L-glutamate | pH 9.0, 40°C, recombinant enzyme | Halobacterium salinarum | |
0.000025 | - |
L-glutamate | pH 9.0, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
0.00013 | - |
NH3 | pH 8.5, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
0.00035 | - |
NH3 | pH 8.5, 40°C, recombinant enzyme | Halobacterium salinarum | |
0.0008 | - |
2-oxoglutarate | pH 8.5, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
0.0034 | - |
2-oxoglutarate | pH 8.5, 40°C, recombinant enzyme | Halobacterium salinarum | |
0.009 | - |
NADH | pH 8.5, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
0.014 | - |
NADH | pH 8.5, 40°C, recombinant enzyme | Halobacterium salinarum | |
0.112 | - |
NAD+ | pH 9.0, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
0.118 | - |
NAD+ | pH 9.0, 40°C, recombinant enzyme | Halobacterium salinarum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
betaine | high concentrations of salt may be substituted with 30% DMSO or betaine with good stability and activity | Halobacterium salinarum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate + H2O + NAD+ | Halobacterium salinarum | - |
2-oxoglutarate + NH3 + NADH + H+ | - |
r | |
L-glutamate + H2O + NAD+ | Halobacterium salinarum NRC-36014 | - |
2-oxoglutarate + NH3 + NADH + H+ | - |
r |
Organic Solvent | Comment | Organism |
---|---|---|
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 |
Acetone | GDHX retains 38% activity in 10% after 72 h, pH 8.5, room temperature | Halobacterium salinarum |
acetonitrile | GDHX retains 30% activity in 10% after 72 h, pH 8.5, room temperature | Halobacterium salinarum |
dimethylformamide | 51% activity remaining after 72 h, pH 8.5, room temperature | Halobacterium salinarum |
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 |
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 |
Ethanol | GDHX retains 34% activity in 10% after 72 h, pH 8.5, room temperature | Halobacterium salinarum |
Glycerol | GDHX retains 73% activity in 10% glycerol after 72 h, pH 8.5, room temperature | Halobacterium salinarum |
Methanol | GDHX retains more than 73% activity in 10% after 72 h, pH 8.5, room temperature | Halobacterium salinarum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Halobacterium salinarum | P29051 | - |
- |
Halobacterium salinarum NRC-36014 | P29051 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant soluble enzyme 11.3fold from Haloferax volcanii strain DS70, by hydrophobic interaction and anion exchange chromatography, method optimization | Halobacterium salinarum |
Renatured (Comment) | Organism |
---|---|
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 Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
6.52 | - |
recombinant enzyme in Haloferax volcanii, pH 8.5, 40°C | Halobacterium salinarum |
35 | - |
recombinant enzyme purified from Haloferax volcanii, pH 8.5, 40°C | Halobacterium salinarum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate + H2O + NAD+ | - |
Halobacterium salinarum | 2-oxoglutarate + NH3 + NADH + H+ | - |
r | |
L-glutamate + H2O + NAD+ | - |
Halobacterium salinarum NRC-36014 | 2-oxoglutarate + NH3 + NADH + H+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
NAD+-dependent GDHX | - |
Halobacterium salinarum |
NAD+-dependent glutamate dehydrogenase | - |
Halobacterium salinarum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
assay at | Halobacterium salinarum |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 90 | temperature profile, overview | Halobacterium salinarum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 70 | recombinant purified enzyme, fully stable at 40°C | Halobacterium salinarum |
80 | - |
recombinant purified enzyme, 30 min, 88% remaining | Halobacterium salinarum |
90 | - |
recombinant purified enzyme, 30 min, 56% remaining | Halobacterium salinarum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
15 | - |
NAD+ | pH 9.0, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
16 | - |
NAD+ | pH 9.0, 40°C, recombinant enzyme | Halobacterium salinarum | |
17 | - |
L-glutamate | pH 9.0, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
24 | - |
L-glutamate | pH 9.0, 40°C, recombinant enzyme | Halobacterium salinarum | |
39 | - |
NADH | pH 8.5, 40°C, recombinant enzyme | Halobacterium salinarum | |
46 | - |
2-oxoglutarate | pH 8.5, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
47 | - |
NADH | pH 8.5, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
56 | - |
2-oxoglutarate | pH 8.5, 40°C, recombinant enzyme | Halobacterium salinarum | |
84 | - |
NH3 | pH 8.5, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
85 | - |
NH3 | pH 8.5, 40°C, recombinant enzyme | Halobacterium salinarum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
amination | Halobacterium salinarum |
9 | - |
deamination | Halobacterium salinarum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.5 | 10.5 | pH profile, overvew | Halobacterium salinarum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Halobacterium salinarum | |
NADH | - |
Halobacterium salinarum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0018 | - |
L-glutamate | pH 9.0, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
0.0035 | - |
L-glutamate | pH 9.0, 40°C, recombinant enzyme | Halobacterium salinarum | |
0.72 | - |
NAD+ | pH 9.0, 40°C, recombinant enzyme | Halobacterium salinarum | |
0.75 | - |
NAD+ | pH 9.0, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
1.7 | - |
NADH | pH 8.5, 40°C, recombinant enzyme | Halobacterium salinarum | |
1.9 | - |
NADH | pH 8.5, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
16.5 | - |
2-oxoglutarate | pH 8.5, 40°C, recombinant enzyme | Halobacterium salinarum | |
45 | - |
NH3 | pH 8.5, 40°C, recombinant enzyme | Halobacterium salinarum | |
57.5 | - |
2-oxoglutarate | pH 8.5, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum | |
115 | - |
NH3 | pH 8.5, 40°C, recombinant enzyme in presence of DMSO | Halobacterium salinarum |