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Literature summary for 6.3.2.3 extracted from

  • Albino, A.; Marco, S.; Di Maro, A.; Chambery, A.; Masullo, M.; De Vendittis, E.
    Characterization of a cold-adapted glutathione synthetase from the psychrophile Pseudoalteromonas haloplanktis (2012), Mol. Biosyst., 8, 2405-2414.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene gshB, located on chromosome I, expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Pseudoalteromonas haloplanktis

Inhibitors

Inhibitors Comment Organism Structure
GSSG the oxidised form of glutathione acts as an irreversible inhibitor of recombinant GshB Pseudoalteromonas haloplanktis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.17
-
ATP pH 7.8, 10°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
0.18
-
ATP pH 7.8, 20°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
0.18
-
ATP pH 7.8, 30°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
0.2 1 ATP pH 7.8, 28°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
0.25
-
gamma-L-glutamyl-L-cysteine pH 7.8, 30°C, recombinant enzyme Pseudoalteromonas haloplanktis
0.26
-
ATP pH 7.8, 15°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
0.26
-
ATP pH 7.8, 30°C, recombinant enzyme, with gamma-L-glutamyl-L-cysteine and glycine Pseudoalteromonas haloplanktis
0.28
-
ATP pH 7.8, 25°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
0.75
-
glycine pH 7.8, 30°C, recombinant enzyme Pseudoalteromonas haloplanktis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ a divalent cation is absolutely required for the activity, whereas monovalent cations are dispensable. Mg2+ is significantly more effective than Mn2+ Pseudoalteromonas haloplanktis
Mn2+ a divalent cation is absolutely required for the activity, whereas monovalent cations are dispensable. Mg2+ is significantly more effective than Mn2+ Pseudoalteromonas haloplanktis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35888
-
2 * 36000, recombinant His-tagged enzyme, SDS-PAGE, 2 * 35888, sequence calculation Pseudoalteromonas haloplanktis
36000
-
2 * 36000, recombinant His-tagged enzyme, SDS-PAGE, 2 * 35888, sequence calculation Pseudoalteromonas haloplanktis
74000
-
recombinant His-tagged enzyme, gel filtration Pseudoalteromonas haloplanktis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + gamma-L-glutamyl-L-cysteine + glycine Pseudoalteromonas haloplanktis
-
ADP + phosphate + glutathione
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine Pseudoalteromonas haloplanktis TAC 125
-
ADP + phosphate + glutathione
-
?
additional information Pseudoalteromonas haloplanktis the enzyme forms a disulfide adduct with 2-mercaptoethanol, when purified in the presence of this reducing agent ?
-
?
additional information Pseudoalteromonas haloplanktis TAC 125 the enzyme forms a disulfide adduct with 2-mercaptoethanol, when purified in the presence of this reducing agent ?
-
?

Organism

Organism UniProt Comment Textmining
Pseudoalteromonas haloplanktis Q3IFA2 cold-adapted, gene gshB
-
Pseudoalteromonas haloplanktis TAC 125 Q3IFA2 cold-adapted, gene gshB
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Pseudoalteromonas haloplanktis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + gamma-L-glutamyl-L-cysteine + glycine
-
Pseudoalteromonas haloplanktis ADP + phosphate + glutathione
-
?
ATP + gamma-L-glutamyl-L-cysteine + glycine
-
Pseudoalteromonas haloplanktis TAC 125 ADP + phosphate + glutathione
-
?
additional information the enzyme forms a disulfide adduct with 2-mercaptoethanol, when purified in the presence of this reducing agent Pseudoalteromonas haloplanktis ?
-
?
additional information assay method development based on [gamma32P]ATP hydrolysis coupled to the GSH synthesis, overview Pseudoalteromonas haloplanktis ?
-
?
additional information the enzyme forms a disulfide adduct with 2-mercaptoethanol, when purified in the presence of this reducing agent Pseudoalteromonas haloplanktis TAC 125 ?
-
?
additional information assay method development based on [gamma32P]ATP hydrolysis coupled to the GSH synthesis, overview Pseudoalteromonas haloplanktis TAC 125 ?
-
?

Subunits

Subunits Comment Organism
homodimer 2 * 36000, recombinant His-tagged enzyme, SDS-PAGE, 2 * 35888, sequence calculation Pseudoalteromonas haloplanktis

Synonyms

Synonyms Comment Organism
gshB
-
Pseudoalteromonas haloplanktis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
-
Pseudoalteromonas haloplanktis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
10 30 the recombinant enzyme displays a great temperature-dependent increase in its activity with an unusually high value of energy of activation of 75kJ/mol for a psychrophilic enzyme Pseudoalteromonas haloplanktis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
-
30 purified recombinant enzyme, pH 7.8, 10 min, completely stable Pseudoalteromonas haloplanktis
50.5
-
purified recombinant enzyme, pH 7.8, 10 min, 50% inactivation Pseudoalteromonas haloplanktis
60
-
purified recombinant enzyme, pH 7.8, 10 min, inactivation Pseudoalteromonas haloplanktis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.87
-
ATP pH 7.8, 10°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
1.85
-
ATP pH 7.8, 15°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
2.42
-
ATP pH 7.8, 20°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
4.72
-
ATP pH 7.8, 25°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
4.99
-
ATP pH 7.8, 28°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
5.32
-
ATP pH 7.8, 30°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4 8.6
-
Pseudoalteromonas haloplanktis

Cofactor

Cofactor Comment Organism Structure
ATP
-
Pseudoalteromonas haloplanktis

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
10.7
-
GSSG pH 7.8, 30°C, recombinant enzyme Pseudoalteromonas haloplanktis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.1
-
ATP pH 7.8, 10°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
7.1
-
ATP pH 7.8, 15°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
13.4
-
ATP pH 7.8, 20°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
16.9
-
ATP pH 7.8, 25°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
23.8
-
ATP pH 7.8, 28°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis
29.6
-
ATP pH 7.8, 30°C, recombinant enzyme, ATPase activity Pseudoalteromonas haloplanktis