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1.4.1.2: glutamate dehydrogenase

This is an abbreviated version!
For detailed information about glutamate dehydrogenase, go to the full flat file.

Word Map on EC 1.4.1.2

Reaction

L-glutamate
+
H2O
+
NAD+
=
2-oxoglutarate
+
NH3
+
NADH
+
H+

Synonyms

At5g18170, AtGDH1, BpNADGDH, c, CCNA_00086, CsGDH, dehydrogenase, glutamate, GDH, GDH isoenzyme 1, GDH, NAD-dependent, gdh-1, gdh-2, GDH1, GDH2, Gdh2p, GDH3, GdhA, GDHB, GDHI, GdhZ, Glu dehydrogenase, GluD, GLUD1, GLUD2, GluDH, glutamate dehydrogenase, glutamate dehydrogenase (NAD), glutamate dehydrogenase 2, glutamate dehydrogenase alpha subunit, glutamate dehydrogenase beta subunit, glutamate dehydrogenase isoform 1, glutamate oxidoreductase, glutamic acid dehydrogenase, glutamic dehydrogenase, hGDH1, hGDH2-nerve-specific GDH, house-keeping GDH, L-glutamate dehydrogenase, L-glutamic acid dehydrogenase, More, NAD(+)-dependent glutamate dehydrogenase, NAD(H)-dependent glutamate dehydrogenase, NAD+-dependant glutamate dehydrogenase, NAD+-dependent GDH, NAD+-dependent GDHX, NAD+-dependent GluDH, NAD+-dependent glutamate dehydrogenase, NAD+-GDH, NAD+-glutamate dehydrogenase, NAD+-specific GDH, NAD+-specific glutamate dehydrogenase, NAD-dependent GDH, NAD-dependent glutamate dehydrogenase, NAD-dependent glutamic dehydrogenase, NAD-dependent L-glutamate dehydrogenase, NAD-GDH, NAD-glutamate dehydrogenase, NAD-linked glutamate dehydrogenase, NAD-linked glutamic dehydrogenase, NAD-specific glutamate dehydrogenase, NAD-specific glutamic dehydrogenase, NAD-ylGdh2p, NAD:glutamate oxidoreductase, NADH-dependent GDH, NADH-dependent glutamate dehydrogenase, NADH-GDH, NADH-glutamate dehydrogenase, NADH-linked glutamate dehydrogenase, OsGDH1, OsGDH2, OsGDH3, Pcal_1031, RocG, sco2999, Surface-associated protein PGAG1, t-GDH, type I GDH, YALI0E09603g, ylGDH2

ECTree

     1 Oxidoreductases
         1.4 Acting on the CH-NH2 group of donors
             1.4.1 With NAD+ or NADP+ as acceptor
                1.4.1.2 glutamate dehydrogenase

Purification

Purification on EC 1.4.1.2 - glutamate dehydrogenase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
18-22°C, 20% glycerol, heat treatment, ammonium sulfate, gel filtration, ion-exchange chromatography
-
2 isoenzymes GDHA, GDHB
-
2 isoenzymes: GDH-I and GDH-II
-
4 isoenzymes
-
7 isoenzymes
-
activated and non-activated form
-
by sonication and centrifugation
DEAE-cellulose column chromatography and Super-Q Toyopearl column chromatography
-
DEAE-Sephacel column chromatography
-
DEAE-Toyopearl column chromatography, butyl-Toyopearl column chromatography, and Superdex 200 gel filtration
GDH I and II
-
GDH is readily adsorbed on highly activated anionic exchangers (HAAE), but hardly adsorbed on lowly activated supports (LAAE) or on highly activated epoxy supports. Using amino-epoxy supports, GDH immobilizes on HAAE-epoxy and more slowly on LAAE-epoxy supports. Both immobilized biocatalysts are incubated at pH 10 for different times to increase the multipoint covalent attachment. Lowly activated anionic exchangers-epoxy-GDH is stable at pH 4 and 25°C, the enzyme stability does not depend on the enzyme concentration and does not release any subunit to the supernatant, in opposition to the results obtained using HAAE-epoxy supports
-
gel filtration
-
gel filtration, native enzyme 232fold purified, recombinant enzyme 5.4fold purified
inactivate recombinant enzyme expressed from Escherichia coli by affinity and anion exchange chromatography, followed by gel filtrration, activation by 5 M urea and 70°c
-
NAD+-dependent and NADP+-dependent GDH
-
NAD-GDH is best obtained from proline grown Aspergillus niger mycelia
-
native enzyme from acetone powder of chicken liver by ammonium sulfate fractionation and dialysis, to near homogeneity (about over 90% purity)
-
Ni2+ HiTrap chelating column chromatography
-
overexpression in Escherichia coli
overexpression of F187D mutant in Escherichia coli
-
overexpression of GDHB in Escherichia coli
-
overexpression of mutants in Escherichia coli
-
Q sepharose column chromatography
-
recombinant enzyme
-
recombinant enzyme from Escherichia coli TOP10 cells by hydrophobic interaction chromatography
recombinant His-tagged enzyme
-
recombinant N-terminally Met-Ala-Ser-tagged and C-terminally His6-tagged enzyme from Eschericia coli strain Rosetta(DE3) by affinity chromatography and gel filtration
-
recombinant soluble enzyme 11.3fold from Haloferax volcanii strain DS70, by hydrophobic interaction and anion exchange chromatography, method optimization
recombinant wild-type and mutant enzymes by affinity chromatography purification, followed by gel filtration
-
recombinant wild-type and mutant enzymes from Escherichia coli strain XL1-Blue
-
Remazol-Red column chromatography
-
to homogeneity
to homogeneity, about 39fold
using dye-affintiy chromatography and anion-exchange chromatography
-