Cloned (Comment) | Organism |
---|---|
gene ggtB, recombinat expression of GgtB with an N-terminal fusion of the chitin-binding domain and intein tag to the target protein, subcloning in Escherichia coli strain DH5alphaMCR and subsequently transformation into and expression in Escherichia coli strain ER25669 | Corynebacterium glutamicum |
Protein Variants | Comment | Organism |
---|---|---|
additional information | deletion of the complete coding sequence of ggtB in the chromosome of Corynebacterium glutamicum. Intracellular concentrations of gamma-glutamyl dipeptides upon deletion and overexpression of gene ggtB in Corynebacterium glutamicum strain ATCC 13032 | Corynebacterium glutamicum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
NaCl | at 10% NaCl, GgtBCg still retains 70% of its initial activity. Further addition of NaCl to concentrations of 15%, 20%, and 22.5% reduces the activity to 30%, 27%, and 20%, respectively. For all tested salt concentrations, activity is higher in the presence of glycyl-glycine than in its absence | Corynebacterium glutamicum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.123 | - |
L-gamma-glutamyl-4-nitroanilide | pH 9.6, 37°C, recombinant enzyme | Corynebacterium glutamicum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cell surface | the enzyme is a lipoprotein and is attached to the outer side of the cytoplasmic membrane | Corynebacterium glutamicum | 9986 | - |
membrane | the enzyme is a lipoprotein and is attached to the outer side of the cytoplasmic membrane | Corynebacterium glutamicum | 16020 | - |
additional information | prediction of signal peptides in protein sequences of the N-terminus with a putative SpI and a SpII cleavage site, overview | Corynebacterium glutamicum | - |
- |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
a (5-L-glutamyl)-peptide + an amino acid | Corynebacterium glutamicum | - |
a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | Corynebacterium glutamicum LMG 3730 | - |
a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | Corynebacterium glutamicum ATCC 13032 | - |
a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | Corynebacterium glutamicum JCM 1318 | - |
a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | Corynebacterium glutamicum NCIMB 10025 | - |
a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | Corynebacterium glutamicum DSM 20300 | - |
a peptide + a 5-L-glutamyl amino acid | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Corynebacterium glutamicum | Q8NRT7 | - |
- |
Corynebacterium glutamicum ATCC 13032 | Q8NRT7 | - |
- |
Corynebacterium glutamicum DSM 20300 | Q8NRT7 | - |
- |
Corynebacterium glutamicum JCM 1318 | Q8NRT7 | - |
- |
Corynebacterium glutamicum LMG 3730 | Q8NRT7 | - |
- |
Corynebacterium glutamicum NCIMB 10025 | Q8NRT7 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
lipoprotein | the enzyme is a lipoprotein and is attached to the outer side of the cytoplasmic membrane | Corynebacterium glutamicum |
proteolytic modification | the enzyme is expressed as a single inactive pre-protein that undergoes posttranslational proteolytic self-cleavage | Corynebacterium glutamicum |
Purification (Comment) | Organism |
---|---|
recombinat GgtB N-terminally tagged with chitin-binding domain and intein tag from Escherichia coli strain ER25669 by chitin affinity chromatography and ultrafiltration | Corynebacterium glutamicum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
quantitation of dipeptides and amino acids by HPLC in whole-cell assays | Corynebacterium glutamicum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
a (5-L-glutamyl)-peptide + an amino acid | - |
Corynebacterium glutamicum | a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | - |
Corynebacterium glutamicum LMG 3730 | a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | - |
Corynebacterium glutamicum ATCC 13032 | a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | - |
Corynebacterium glutamicum JCM 1318 | a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | - |
Corynebacterium glutamicum NCIMB 10025 | a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | - |
Corynebacterium glutamicum DSM 20300 | a peptide + a 5-L-glutamyl amino acid | - |
? | |
L-gamma-glutamyl-4-nitroanilide + glycylglycine | - |
Corynebacterium glutamicum | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + glycylglycine | - |
Corynebacterium glutamicum LMG 3730 | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + glycylglycine | - |
Corynebacterium glutamicum ATCC 13032 | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + glycylglycine | - |
Corynebacterium glutamicum JCM 1318 | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + glycylglycine | - |
Corynebacterium glutamicum NCIMB 10025 | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + glycylglycine | - |
Corynebacterium glutamicum DSM 20300 | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamate | - |
Corynebacterium glutamicum | 4-nitroaniline + 5-L-glutamyl-L-glutamate | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamate | - |
Corynebacterium glutamicum LMG 3730 | 4-nitroaniline + 5-L-glutamyl-L-glutamate | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamate | - |
Corynebacterium glutamicum ATCC 13032 | 4-nitroaniline + 5-L-glutamyl-L-glutamate | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamate | - |
Corynebacterium glutamicum JCM 1318 | 4-nitroaniline + 5-L-glutamyl-L-glutamate | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamate | - |
Corynebacterium glutamicum NCIMB 10025 | 4-nitroaniline + 5-L-glutamyl-L-glutamate | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamate | - |
Corynebacterium glutamicum DSM 20300 | 4-nitroaniline + 5-L-glutamyl-L-glutamate | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamine | - |
Corynebacterium glutamicum | 4-nitroaniline + 5-L-glutamyl-L-glutamine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamine | - |
Corynebacterium glutamicum LMG 3730 | 4-nitroaniline + 5-L-glutamyl-L-glutamine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamine | - |
Corynebacterium glutamicum ATCC 13032 | 4-nitroaniline + 5-L-glutamyl-L-glutamine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamine | - |
Corynebacterium glutamicum JCM 1318 | 4-nitroaniline + 5-L-glutamyl-L-glutamine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamine | - |
Corynebacterium glutamicum NCIMB 10025 | 4-nitroaniline + 5-L-glutamyl-L-glutamine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-glutamine | - |
Corynebacterium glutamicum DSM 20300 | 4-nitroaniline + 5-L-glutamyl-L-glutamine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-leucine | - |
Corynebacterium glutamicum | 4-nitroaniline + 5-L-glutamyl-L-leucine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-methionine | - |
Corynebacterium glutamicum | 4-nitroaniline + 5-L-glutamyl-L-methionine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + L-valine | - |
Corynebacterium glutamicum | 4-nitroaniline + 5-L-glutamyl-L-valine | - |
? | |
additional information | L-gamma-glutamyl-4-nitroanilide is a chromogenic substrate. With the isolated recombinant enzyme, it is determined that besides GPNA also the dipeptides gamma-Glu-Glu, gamma-Glu-Gln, gamma-Glu-Met, gamma-Glu-Val, and gamma-Glu-Leu are hydrolyzed, cf. EC 3.4.19.13 | Corynebacterium glutamicum | ? | - |
- |
|
additional information | L-gamma-glutamyl-4-nitroanilide is a chromogenic substrate. With the isolated recombinant enzyme, it is determined that besides GPNA also the dipeptides gamma-Glu-Glu, gamma-Glu-Gln, gamma-Glu-Met, gamma-Glu-Val, and gamma-Glu-Leu are hydrolyzed, cf. EC 3.4.19.13 | Corynebacterium glutamicum LMG 3730 | ? | - |
- |
|
additional information | L-gamma-glutamyl-4-nitroanilide is a chromogenic substrate. With the isolated recombinant enzyme, it is determined that besides GPNA also the dipeptides gamma-Glu-Glu, gamma-Glu-Gln, gamma-Glu-Met, gamma-Glu-Val, and gamma-Glu-Leu are hydrolyzed, cf. EC 3.4.19.13 | Corynebacterium glutamicum ATCC 13032 | ? | - |
- |
|
additional information | L-gamma-glutamyl-4-nitroanilide is a chromogenic substrate. With the isolated recombinant enzyme, it is determined that besides GPNA also the dipeptides gamma-Glu-Glu, gamma-Glu-Gln, gamma-Glu-Met, gamma-Glu-Val, and gamma-Glu-Leu are hydrolyzed, cf. EC 3.4.19.13 | Corynebacterium glutamicum JCM 1318 | ? | - |
- |
|
additional information | L-gamma-glutamyl-4-nitroanilide is a chromogenic substrate. With the isolated recombinant enzyme, it is determined that besides GPNA also the dipeptides gamma-Glu-Glu, gamma-Glu-Gln, gamma-Glu-Met, gamma-Glu-Val, and gamma-Glu-Leu are hydrolyzed, cf. EC 3.4.19.13 | Corynebacterium glutamicum NCIMB 10025 | ? | - |
- |
|
additional information | L-gamma-glutamyl-4-nitroanilide is a chromogenic substrate. With the isolated recombinant enzyme, it is determined that besides GPNA also the dipeptides gamma-Glu-Glu, gamma-Glu-Gln, gamma-Glu-Met, gamma-Glu-Val, and gamma-Glu-Leu are hydrolyzed, cf. EC 3.4.19.13 | Corynebacterium glutamicum DSM 20300 | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
heterodimer | bacterial gamma-glutamyl transpeptidase (GGT) is a heterodimeric enzyme, consisting of one large and one small subunit | Corynebacterium glutamicum |
Synonyms | Comment | Organism |
---|---|---|
Cgl0954 | - |
Corynebacterium glutamicum |
gamma-glutamyl transpeptidase | - |
Corynebacterium glutamicum |
ggtB | - |
Corynebacterium glutamicum |
More | see also EC 3.4.19.13 | Corynebacterium glutamicum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
- |
Corynebacterium glutamicum |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | 60 | the optimal temperature of GgtBCg is 45°C. The enzyme exhibits 85% and 60% of the maximal activity at 37°C and 22°C, respectively. Whereas GgtBCg retains 50% activity at 60°C, it is completely inactivated at 70°C, recombinant enzyme | Corynebacterium glutamicum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9.6 | - |
- |
Corynebacterium glutamicum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 11 | activity range, recombinant enzyme | Corynebacterium glutamicum |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the superfamily of N-terminal nucleophile (NTN-)aminohydrolases | Corynebacterium glutamicum |
malfunction | gamma-glutamyl transpeptidase activity is abolished upon deletion of ggtB. But although deletion and overexpression of ggtB has significant effects on intracellular dipeptide concentrations, it is neither essential for biosynthesis nor catabolismof these dipeptides in vivo | Corynebacterium glutamicum |
physiological function | gamma-glutamyl transpeptidase activity is associated with intact cells of Corynebacterium glutamicum. Enzyme GgtB catalyzes the concentration-dependent synthesis and hydrolysis of gamma-glutamyl dipeptides and shows strong glutaminase activity. The intracellular concentrations of five gamma-glutamyl dipeptides, i.e. gamma-Glu-Glu, gamma-Glu-Gln, gamma-Glu-Val, gamma-Glu-Leu, gamma-Glu-Met, are determined by HPLC-MS. Metabolic profiling reveals that ggtB is not essential for theformation of intracellular gamma-glutamyl dipeptides. GgtBCg might contribute to catabolism by hydrolyzing L-glutamine to glutamate and ammonia | Corynebacterium glutamicum |