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Literature summary for 1.4.1.4 extracted from

  • Vamsee-Krishna, C.; Phale, P.S.
    Bypassing isophthalate inhibition by modulating glutamate dehydrogenase (GDH): purification and kinetic characterization of NADP-GDHs from isophthalate-degrading Pseudomonas aeruginosa strain PP4 and Acinetobacter lwoffii strain ISP4 (2010), J. Bacteriol., 192, 801-806.
    View publication on PubMedView publication on EuropePMC

Inhibitors

Inhibitors Comment Organism Structure
isophthalate a competitive inhibitor of glutamate dehydrogenase, is involved in C and N metabolism Acinetobacter lwoffii
isophthalate a competitive inhibitor of glutamate dehydrogenase, is involved in C and N metabolism Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetic analysis, overview, the isozyme GDHI' shows negative cooperativity with NH4+ substrate, with a decreased affinity for NH4Cl Acinetobacter lwoffii
additional information
-
additional information kinetic analysis, overview, the isozymes show negative cooperativity with NH4+ substrate, with a decreased affinity for NH4Cl Pseudomonas aeruginosa

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
43000
-
4 * 43000, isozyme GDHI, SDS-PAGE Acinetobacter lwoffii
43000
-
4 * 43000, isozymes GDHI and GDHII, SDS-PAGE Pseudomonas aeruginosa
170000
-
isozymes GDHI and GDHII, gel filtration Pseudomonas aeruginosa
250000
-
isozyme GDHI', gel filtration Acinetobacter lwoffii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-glutamate + H2O + NADP+ Pseudomonas aeruginosa
-
2-oxoglutarate + NH3 + NADPH + H+
-
r
L-glutamate + H2O + NADP+ Acinetobacter lwoffii
-
2-oxoglutarate + NH3 + NADPH + H+
-
r
L-glutamate + H2O + NADP+ Pseudomonas aeruginosa PP4
-
2-oxoglutarate + NH3 + NADPH + H+
-
r
L-glutamate + H2O + NADP+ Acinetobacter lwoffii ISP4
-
2-oxoglutarate + NH3 + NADPH + H+
-
r

Organism

Organism UniProt Comment Textmining
Acinetobacter lwoffii
-
NADPH-GDH isozymes GDHI, GDHII, and GDHI'
-
Acinetobacter lwoffii ISP4
-
NADPH-GDH isozymes GDHI, GDHII, and GDHI'
-
Pseudomonas aeruginosa
-
NADPH-GDH isozymes GDHI, GDHII, and GDHI'
-
Pseudomonas aeruginosa PP4
-
NADPH-GDH isozymes GDHI, GDHII, and GDHI'
-

Purification (Commentary)

Purification (Comment) Organism
native GDHI' 142fold from cells grown on isophthalate and 510fold from cells grown on 2YT Acinetobacter lwoffii
native isozymes GDHI and GDHII from strain PP4 grown on isophthalate and glucose, respectively. Enzyme yields are better in the presence of 1 mM 2-oxoglutarate, 1 mM EDTA, and 4.35% glycerol. GDHI and GDHII are purified 228fold and 428fold, respectively Pseudomonas aeruginosa

Source Tissue

Source Tissue Comment Organism Textmining
culture condition:isophthalate-grown cell isophthalate as a sole source of carbon and energy. Strain ISP4 produces a single form of NADP-GDH, GDHI' when it is grown on either isophthalate or rich medium 2YT Acinetobacter lwoffii
-
culture condition:isophthalate-grown cell isophthalate as a sole source of carbon and energy. Strain PP4 shows carbon source-dependent modulation of NADP-GDH. GDHI is produced when cells are grown on isophthalate Pseudomonas aeruginosa
-

Storage Stability

Storage Stability Organism
4°C, purified enzyme, stable for about 5 to 6 months without any significant loss of activity Acinetobacter lwoffii
4°C, purified enzymes, stable for about 5 to 6 months without any significant loss of activity Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-glutamate + H2O + NADP+
-
Pseudomonas aeruginosa 2-oxoglutarate + NH3 + NADPH + H+
-
r
L-glutamate + H2O + NADP+
-
Acinetobacter lwoffii 2-oxoglutarate + NH3 + NADPH + H+
-
r
L-glutamate + H2O + NADP+
-
Pseudomonas aeruginosa 2-oxoglutarate + NH3 + NADPH + H+ usage of NH4Cl as substrate for the reverse reaction r
L-glutamate + H2O + NADP+ usage of NH4Cl as substrate for the reverse reaction Acinetobacter lwoffii 2-oxoglutarate + NH3 + NADPH + H+
-
r
L-glutamate + H2O + NADP+
-
Pseudomonas aeruginosa PP4 2-oxoglutarate + NH3 + NADPH + H+
-
r
L-glutamate + H2O + NADP+
-
Pseudomonas aeruginosa PP4 2-oxoglutarate + NH3 + NADPH + H+ usage of NH4Cl as substrate for the reverse reaction r
L-glutamate + H2O + NADP+
-
Acinetobacter lwoffii ISP4 2-oxoglutarate + NH3 + NADPH + H+
-
r
L-glutamate + H2O + NADP+ usage of NH4Cl as substrate for the reverse reaction Acinetobacter lwoffii ISP4 2-oxoglutarate + NH3 + NADPH + H+
-
r

Subunits

Subunits Comment Organism
homohexamer 4 * 43000, isozyme GDHI', SDS-PAGE Acinetobacter lwoffii
homotetramer 4 * 43000, isozymes GDHI and GDHII, SDS-PAGE Pseudomonas aeruginosa

Synonyms

Synonyms Comment Organism
GDHI
-
Pseudomonas aeruginosa
GDHI'
-
Acinetobacter lwoffii
NADP-GDH
-
Pseudomonas aeruginosa
NADP-GDH
-
Acinetobacter lwoffii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8 8.5 reverse reaction, assay at Pseudomonas aeruginosa
8 8.5 reverse reaction, assay at Acinetobacter lwoffii
9.5
-
forward reaction, assay at Pseudomonas aeruginosa
9.5
-
forward reaction, assay at Acinetobacter lwoffii

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Pseudomonas aeruginosa
NADP+
-
Acinetobacter lwoffii
NADPH
-
Pseudomonas aeruginosa
NADPH
-
Acinetobacter lwoffii

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
additional information
-
additional information Ki values for isophthalate with different isozymes from differently grown strains in forward and reverse reactions, overview Pseudomonas aeruginosa
additional information
-
additional information Ki values for isophthalate with isozyme from differently grown strains in forward and reverse reactions, overview Acinetobacter lwoffii