Cloned (Comment) | Organism |
---|---|
gene ggt, sequence comparisons, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain M15 | Bacillus licheniformis |
Protein Variants | Comment | Organism |
---|---|---|
additional information | molecular modeling of mutant enzymes using the X-ray crystal structures of BlGGT (PDB ID 4OTT) and T399A-BlGGT (PDB ID 4Y23) as templates. GdnHCl-induced denaturation of BlGGT and its variants, overview | Bacillus licheniformis |
N450A | site-directed mutagenesis, the mutant shows about 4.7fold increased catalytic efficiency compared to wild-type | Bacillus licheniformis |
N450D | site-directed mutagenesis, the mutant shows about 8fold increased catalytic efficiency compared to wild-type | Bacillus licheniformis |
N450K | site-directed mutagenesis, N450K exhibits 81% increase in KM and 44.3% decrease in kcat compared to wild-type, leading to a profound reduction in its catalytic efficiency | Bacillus licheniformis |
N450R | site-directed mutagenesis, the mutant shows a significant reduction in the catalytic activity compared to wild-type | Bacillus licheniformis |
T399A | site-directed mutagenesis, mutant structure analysis and comparison | Bacillus licheniformis |
General Stability | Organism |
---|---|
GdnHCl-induced denaturation of BlGGT and its variants, overview | Bacillus licheniformis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.42 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant wild-type enzyme, pH 9.0, 40°C | Bacillus licheniformis | |
0.58 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant N450D, pH 9.0, 40°C | Bacillus licheniformis | |
0.58 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant N450Q, pH 9.0, 40°C | Bacillus licheniformis | |
0.69 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant N450A, pH 9.0, 40°C | Bacillus licheniformis | |
0.76 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant N450K, pH 9.0, 40°C | Bacillus licheniformis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
a (5-L-glutamyl)-peptide + an amino acid | Bacillus licheniformis | - |
a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | Bacillus licheniformis DSM 13 | - |
a peptide + a 5-L-glutamyl amino acid | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus licheniformis | Q62WE3 | - |
- |
Bacillus licheniformis DSM 13 | Q62WE3 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
proteolytic modification | the enzyme is synthetized as single-chain precursor, and then self-processes to form the mature enzyme. Analysis of autocatalytic processing of recombinant mutant enzymes expressed in Escherichia coli strain M15 | Bacillus licheniformis |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain M15 by nickel affinity chromatography | Bacillus licheniformis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
a (5-L-glutamyl)-peptide + an amino acid | - |
Bacillus licheniformis | a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | the binding of L-glutamate occurs in a concave site lined by residues Arg109,Thr399, Glu438, Asp441, Ser460, Ser461, Gly481, and Gly482 | Bacillus licheniformis | a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | - |
Bacillus licheniformis DSM 13 | a peptide + a 5-L-glutamyl amino acid | - |
? | |
a (5-L-glutamyl)-peptide + an amino acid | the binding of L-glutamate occurs in a concave site lined by residues Arg109,Thr399, Glu438, Asp441, Ser460, Ser461, Gly481, and Gly482 | Bacillus licheniformis DSM 13 | a peptide + a 5-L-glutamyl amino acid | - |
? | |
L-gamma-glutamyl-4-nitroanilide + glycylglycine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
L-gamma-glutamyl-4-nitroanilide + glycylglycine | - |
Bacillus licheniformis DSM 13 | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterodimer | in the crystal structure of BlGGT, the large subunit contains a globular domain consisting of 14 alpha-helices, six small 310 helices and 11 beta-strands, and the small subunit comprises of 3 alpha-helices, two small 310 helices and 11 beta-strands. Furthermore, both subunits provide strands to a nearly flat beta-sheet that constitutes the core of the heterodimeric arrangement. Unlike their structural consistency, the primary structure of GGT enzymes displays a great degree of variability | Bacillus licheniformis |
Synonyms | Comment | Organism |
---|---|---|
BlGGT | - |
Bacillus licheniformis |
gamma-glutamyltranspeptidase | - |
Bacillus licheniformis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
- |
Bacillus licheniformis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
8.93 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant N450K, pH 9.0, 40°C | Bacillus licheniformis | |
16.04 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant wild-type enzyme, pH 9.0, 40°C | Bacillus licheniformis | |
21.74 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant N450Q, pH 9.0, 40°C | Bacillus licheniformis | |
123.65 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant N450A, pH 9.0, 40°C | Bacillus licheniformis | |
175.52 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant N450D, pH 9.0, 40°C | Bacillus licheniformis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Bacillus licheniformis |
General Information | Comment | Organism |
---|---|---|
evolution | Bacillus licheniformis gamma-glutamyltranspeptidase (BlGGT) belongs to N-terminal nucleophile hydrolase superfamily in which all inclusive members are synthetized as single-chain precursors, and then self-processed to form mature enzymes | Bacillus licheniformis |
additional information | the binding of L-glutamate occurs in a concave site lined by residues Arg109,Thr399, Glu438, Asp441, Ser460, Ser461, Gly481, and Gly482. Asn450 interacts with Asp441 by a hydrogenbond and consequently binds with Arg109 and Glu438 through the hydrogen bond network to hold the substrate in the properposition, suggesting that this residue can be also important for the catalytic function of BlGGT. The putative active residue is Thr399. Superimposition of the catalytic environment of wild-type and mutant enzymes, overview | Bacillus licheniformis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
11.75 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant N450K, pH 9.0, 40°C | Bacillus licheniformis | |
37.48 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant N450Q, pH 9.0, 40°C | Bacillus licheniformis | |
38.19 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant wild-type enzyme, pH 9.0, 40°C | Bacillus licheniformis | |
179.02 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant N450A, pH 9.0, 40°C | Bacillus licheniformis | |
302.62 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant N450D, pH 9.0, 40°C | Bacillus licheniformis |