EC Number | Cloned (Comment) | Organism |
---|---|---|
2.3.2.2 | gene ggt, sequence comparisons, recombinant expression of Strep-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Bacillus licheniformis |
3.4.19.13 | expression in Escherichia coli BL21 (DE3) | Bacillus licheniformis |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.3.2.2 | R109E | site-directed mutagenesis, the mutation reduces the enzyme's affinity for L-Gln | Bacillus licheniformis |
2.3.2.2 | R109F | site-directed mutagenesis, the mutation reduces the enzyme's affinity for L-Gln | Bacillus licheniformis |
2.3.2.2 | R109K | site-directed mutagenesis of the catalytic residue results in low enzyme consumption, the mutation reduces the enzyme's affinity for L-Gln. The Arg109Lys mutant has increased transpeptidation activity and catalytic efficiency compared to the other R109 mutants. The Arg109Lys mutant shows high conversion rates for L-theanine synthesis as well | Bacillus licheniformis |
2.3.2.2 | R109L | site-directed mutagenesis, the mutation reduces the enzyme's affinity for L-Gln, highly reduced transpeptidase activity compared to wild-type | Bacillus licheniformis |
2.3.2.2 | R109M | site-directed mutagenesis of the catalytic residue results in reduced activity and high salt stability of the mutant, the mutation reduces the enzyme's affinity for L-Gln. The Arg109Met mutant shows increased hydrolytic activity as it completely alters the binding of L-Gln at the active site. Also, the salt stability of the mutant enzyme is significantly improved. Highly reduced transpeptidase activity compared to wild-type | Bacillus licheniformis |
2.3.2.2 | R109S | site-directed mutagenesis, the mutation reduces the enzyme's affinity for L-Gln | Bacillus licheniformis |
EC Number | General Stability | Organism |
---|---|---|
3.4.19.13 | mutant enzyme R109M shows high salt stability with almost no loss in activity even in the presence of 20% (w/v) NaCl whereas wild-type enzyme and mutant enzyme R109K retain only 50% of residual activity | Bacillus licheniformis |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
2.3.2.2 | NaCl | - |
Bacillus licheniformis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.3.2.2 | additional information | - |
additional information | enzyme kinetics analysis is done according to the ping-pong mechanism when both the substrates L-gamma-glutamyl-4-nitroanilide and glycyl-glycine are used | Bacillus licheniformis | |
2.3.2.2 | 0.3 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109F, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 0.4 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109E, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 0.4 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109S, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 0.4 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant wild-type enzyme, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 0.7 | - |
glycylglycine | recombinant mutant R109E, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 0.7 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109K, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 0.8 | - |
glycylglycine | recombinant mutant R109F, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 0.94 | - |
glycylglycine | recombinant wild-type enzyme, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 1.5 | - |
glycylglycine | recombinant mutant R109K, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 1.6 | - |
glycylglycine | recombinant mutant R109S, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 8.2 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109L, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 10.6 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109M, pH 9.0, 60°C | Bacillus licheniformis |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.3.2.2 | a (5-L-glutamyl)-peptide + an amino acid | Bacillus licheniformis | - |
a peptide + a 5-L-glutamyl amino acid | - |
? | |
2.3.2.2 | a (5-L-glutamyl)-peptide + an amino acid | Bacillus licheniformis ER-15 | - |
a peptide + a 5-L-glutamyl amino acid | - |
? | |
2.3.2.2 | a (5-L-glutamyl)-peptide + an amino acid | Bacillus licheniformis ER15 | - |
a peptide + a 5-L-glutamyl amino acid | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.3.2.2 | Bacillus licheniformis | A9YTT0 | - |
- |
2.3.2.2 | Bacillus licheniformis ER15 | A9YTT0 | - |
- |
3.4.19.13 | Bacillus licheniformis | A9YTT0 | - |
- |
3.4.19.13 | Bacillus licheniformis ER-15 | A9YTT0 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.3.2.2 | recombinant Strep-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by affinity chromatography and ultrafiltration, to homogeneity | Bacillus licheniformis |
3.4.19.13 | - |
Bacillus licheniformis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.3.2.2 | a (5-L-glutamyl)-peptide + an amino acid | - |
Bacillus licheniformis | a peptide + a 5-L-glutamyl amino acid | - |
? | |
2.3.2.2 | a (5-L-glutamyl)-peptide + an amino acid | - |
Bacillus licheniformis ER-15 | a peptide + a 5-L-glutamyl amino acid | - |
? | |
2.3.2.2 | a (5-L-glutamyl)-peptide + an amino acid | - |
Bacillus licheniformis ER15 | a peptide + a 5-L-glutamyl amino acid | - |
? | |
2.3.2.2 | L-gamma-glutamyl-4-nitroanilide + glycylglycine | - |
Bacillus licheniformis | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
2.3.2.2 | L-gamma-glutamyl-4-nitroanilide + glycylglycine | - |
Bacillus licheniformis ER-15 | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
2.3.2.2 | L-gamma-glutamyl-4-nitroanilide + glycylglycine | - |
Bacillus licheniformis ER15 | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
2.3.2.2 | L-glutamine + ethylamine | formation of gamma-glutamylethylamine (L-theanine) | Bacillus licheniformis | NH3 + gamma-glutamylethylamine | - |
? | |
2.3.2.2 | L-glutamine + ethylamine | formation of gamma-glutamylethylamine (L-theanine) | Bacillus licheniformis ER-15 | NH3 + gamma-glutamylethylamine | - |
? | |
2.3.2.2 | L-glutamine + ethylamine | formation of gamma-glutamylethylamine (L-theanine) | Bacillus licheniformis ER15 | NH3 + gamma-glutamylethylamine | - |
? | |
2.3.2.2 | additional information | GGT is known to catalyze three types of reactions: (a) transpeptidation- when the acceptor is an NH2- group containing compound to which GGT can transfer the gamma-glutamyl moiety, (b) hydrolysis- when gamma-glutamyl moiety is transferred to a water molecule and glutamic acid is released as a product and (c) auto-transpeptidation- when the free NH2- group of a donor acts as an acceptor itself. Amino acid acceptor substrate specificity, overview | Bacillus licheniformis | ? | - |
- |
|
2.3.2.2 | additional information | GGT is known to catalyze three types of reactions: (a) transpeptidation- when the acceptor is an NH2- group containing compound to which GGT can transfer the gamma-glutamyl moiety, (b) hydrolysis- when gamma-glutamyl moiety is transferred to a water molecule and glutamic acid is released as a product and (c) auto-transpeptidation- when the free NH2- group of a donor acts as an acceptor itself. Amino acid acceptor substrate specificity, overview | Bacillus licheniformis ER-15 | ? | - |
- |
|
2.3.2.2 | additional information | GGT is known to catalyze three types of reactions: (a) transpeptidation- when the acceptor is an NH2- group containing compound to which GGT can transfer the gamma-glutamyl moiety, (b) hydrolysis- when gamma-glutamyl moiety is transferred to a water molecule and glutamic acid is released as a product and (c) auto-transpeptidation- when the free NH2- group of a donor acts as an acceptor itself. Amino acid acceptor substrate specificity, overview | Bacillus licheniformis ER15 | ? | - |
- |
|
3.4.19.13 | gamma-glutamyl-p-nitroanilide + H2O | hydrolytic activity | Bacillus licheniformis | L-glutamate + p-nitroaniline | - |
? | |
3.4.19.13 | additional information | the enzyme also shows transpeptidase activity | Bacillus licheniformis | ? | - |
? | |
3.4.19.13 | additional information | the enzyme also shows transpeptidase activity | Bacillus licheniformis ER-15 | ? | - |
? | |
3.4.19.13 | additional information | the enzyme also shows transpeptidase activity | Bacillus licheniformis ER15 | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
2.3.2.2 | heterodimer | 1 * 45000, large subunit, + 1 * 22000, small subunit, His6-tagged enzyme, SDS-PAGE | Bacillus licheniformis |
3.4.19.13 | ? | x * 45000 + x * 22000, SDS-PAGE, all the enzyme variants are of the same molecular weight with large subunit located around 45 kDa and small subunit at 22 kDa | Bacillus licheniformis |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
2.3.2.2 | BlGGT | - |
Bacillus licheniformis |
2.3.2.2 | gamma-glutamyl transpeptidase | - |
Bacillus licheniformis |
2.3.2.2 | GGT | - |
Bacillus licheniformis |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
2.3.2.2 | 60 | - |
recombinant enzyme mutants R109K and R109M | Bacillus licheniformis |
2.3.2.2 | 70 | - |
recombinant wild-type enzyme | Bacillus licheniformis |
3.4.19.13 | 60 | - |
optimum of wild-type enzyme, mutant enzyme R109K and R109M | Bacillus licheniformis |
3.4.19.13 | 70 | - |
optimum of mutant enzyme R109S | Bacillus licheniformis |
EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.4.19.13 | 50 | - |
t1/2: 176 min (mutant enzyme R109M), 16.5 min (mutant enzyme R109S), 69.3 min (wild-type enzyme), 19.8 min (mutant enzyme R109K) | Bacillus licheniformis |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.3.2.2 | 1.7 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109E, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 1.7 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109F, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 8.3 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109L, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 21.7 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109M, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 95 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109S, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 106.7 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant wild-type enzyme, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 305 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109K, pH 9.0, 60°C | Bacillus licheniformis |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
2.3.2.2 | 8 | - |
recombinant enzyme mutant R109M | Bacillus licheniformis |
2.3.2.2 | 9 | - |
recombinant enzyme mutants R109S and R109K | Bacillus licheniformis |
2.3.2.2 | 9 | 10 | recombinant wild-type enzyme | Bacillus licheniformis |
3.4.19.13 | 8 | - |
pH-optimum of mutant enzyme R109M | Bacillus licheniformis |
3.4.19.13 | 9 | - |
comparable activities at both pH 9.0 and pH 10.0. A similar pattern is observed for R109S. Mutant enzyme R109K shows a sharp pH-optimum at pH 9.0 | Bacillus licheniformis |
3.4.19.13 | 10 | - |
comparable activities at both pH 9.0 and pH 10.0. A similar pattern is observed for R109S | Bacillus licheniformis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
2.3.2.2 | evolution | gamma-glutamyl transpeptidase enzyme (GGT) is a member of the N-terminal nucleophile hydrolase superfamily | Bacillus licheniformis |
2.3.2.2 | additional information | GGT enzymes follow a ping-pong mechanism of enzyme catalysis wherein the second substrate binds only when the first substrate is already bound to the enzyme and forms an enzyme-substrate complex. The C-alpha carboxyl group interacts with Arg109, Ser460 and Ser461 and the C-alpha amino group forms contacts with Glu438 and Asp441 while the side chain carboxylic group is oriented towards Thr399, Gly481, and Gly482. Arg109 interacts with C-alpha carboxyl group of the substrate | Bacillus licheniformis |
2.3.2.2 | physiological function | gamma-glutamyl transpeptidase (GGT) catalyzes the transfer of the gamma-glutamyl moiety from donor compounds such as L-glutamine (Gln) and glutathione (GSH) to an acceptor | Bacillus licheniformis |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.3.2.2 | 1.01 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109L, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 2.05 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109M, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 4.25 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109E, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 5.67 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109F, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 237.5 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109S, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 266.75 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant wild-type enzyme, pH 9.0, 60°C | Bacillus licheniformis | |
2.3.2.2 | 435.7 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109K, pH 9.0, 60°C | Bacillus licheniformis |