Any feedback?
Literature summary for 2.3.2.2 extracted from
- Evolving transpeptidase and hydrolytic variants of gamma-glutamyl transpeptidase from Bacillus licheniformis by targeted mutations of conserved residue Arg109 and their biotechnological relevance (2017), J. Biotechnol., 249, 82-90 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene ggt, sequence comparisons, recombinant expression of Strep-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Bacillus licheniformis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| R109E | site-directed mutagenesis, the mutation reduces the enzyme's affinity for L-Gln | Bacillus licheniformis |
| R109F | site-directed mutagenesis, the mutation reduces the enzyme's affinity for L-Gln | Bacillus licheniformis |
| 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 |
| R109L | site-directed mutagenesis, the mutation reduces the enzyme's affinity for L-Gln, highly reduced transpeptidase activity compared to wild-type | Bacillus licheniformis |
| 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 |
| R109S | site-directed mutagenesis, the mutation reduces the enzyme's affinity for L-Gln | Bacillus licheniformis |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| NaCl | - |
Bacillus licheniformis |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 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 | |
| 0.3 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109F, pH 9.0, 60°C | Bacillus licheniformis | |
| 0.4 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109E, pH 9.0, 60°C | Bacillus licheniformis | |
| 0.4 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109S, pH 9.0, 60°C | Bacillus licheniformis | |
| 0.4 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant wild-type enzyme, pH 9.0, 60°C | Bacillus licheniformis | |
| 0.7 | - |
glycylglycine | recombinant mutant R109E, pH 9.0, 60°C | Bacillus licheniformis | |
| 0.7 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109K, pH 9.0, 60°C | Bacillus licheniformis | |
| 0.8 | - |
glycylglycine | recombinant mutant R109F, pH 9.0, 60°C | Bacillus licheniformis | |
| 0.94 | - |
glycylglycine | recombinant wild-type enzyme, pH 9.0, 60°C | Bacillus licheniformis | |
| 1.5 | - |
glycylglycine | recombinant mutant R109K, pH 9.0, 60°C | Bacillus licheniformis | |
| 1.6 | - |
glycylglycine | recombinant mutant R109S, pH 9.0, 60°C | Bacillus licheniformis | |
| 8.2 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109L, pH 9.0, 60°C | Bacillus licheniformis | |
| 10.6 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109M, pH 9.0, 60°C | Bacillus licheniformis | |
Natural Substrates/ Products (Substrates)
| 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 ER-15 | - |
a peptide + a 5-L-glutamyl amino acid | - |
? | |
| a (5-L-glutamyl)-peptide + an amino acid | Bacillus licheniformis ER15 | - |
a peptide + a 5-L-glutamyl amino acid | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bacillus licheniformis | A9YTT0 | - |
- |
| Bacillus licheniformis ER15 | A9YTT0 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant Strep-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by affinity chromatography and ultrafiltration, to homogeneity | Bacillus licheniformis |
Substrates and Products (Substrate)
| 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 | - |
Bacillus licheniformis ER-15 | a peptide + a 5-L-glutamyl amino acid | - |
? | |
| a (5-L-glutamyl)-peptide + an amino acid | - |
Bacillus licheniformis ER15 | 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 ER-15 | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
| L-gamma-glutamyl-4-nitroanilide + glycylglycine | - |
Bacillus licheniformis ER15 | 4-nitroaniline + 5-L-glutamyl-glycylglycine | - |
? | |
| L-glutamine + ethylamine | formation of gamma-glutamylethylamine (L-theanine) | Bacillus licheniformis | NH3 + gamma-glutamylethylamine | - |
? | |
| L-glutamine + ethylamine | formation of gamma-glutamylethylamine (L-theanine) | Bacillus licheniformis ER-15 | NH3 + gamma-glutamylethylamine | - |
? | |
| L-glutamine + ethylamine | formation of gamma-glutamylethylamine (L-theanine) | Bacillus licheniformis ER15 | NH3 + gamma-glutamylethylamine | - |
? | |
| 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 | ? | - |
- |
|
| 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 | ? | - |
- |
|
| 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 | ? | - |
- |
|
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| heterodimer | 1 * 45000, large subunit, + 1 * 22000, small subunit, His6-tagged enzyme, SDS-PAGE | Bacillus licheniformis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| BlGGT | - |
Bacillus licheniformis |
| gamma-glutamyl transpeptidase | - |
Bacillus licheniformis |
| GGT | - |
Bacillus licheniformis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | - |
recombinant enzyme mutants R109K and R109M | Bacillus licheniformis |
| 70 | - |
recombinant wild-type enzyme | Bacillus licheniformis |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.7 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109E, pH 9.0, 60°C | Bacillus licheniformis | |
| 1.7 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109F, pH 9.0, 60°C | Bacillus licheniformis | |
| 8.3 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109L, pH 9.0, 60°C | Bacillus licheniformis | |
| 21.7 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109M, pH 9.0, 60°C | Bacillus licheniformis | |
| 95 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109S, pH 9.0, 60°C | Bacillus licheniformis | |
| 106.7 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant wild-type enzyme, pH 9.0, 60°C | Bacillus licheniformis | |
| 305 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109K, pH 9.0, 60°C | Bacillus licheniformis | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
recombinant enzyme mutant R109M | Bacillus licheniformis |
| 9 | - |
recombinant enzyme mutants R109S and R109K | Bacillus licheniformis |
| 9 | 10 | recombinant wild-type enzyme | Bacillus licheniformis |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | gamma-glutamyl transpeptidase enzyme (GGT) is a member of the N-terminal nucleophile hydrolase superfamily | Bacillus licheniformis |
| 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 |
| 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 |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.01 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109L, pH 9.0, 60°C | Bacillus licheniformis | |
| 2.05 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109M, pH 9.0, 60°C | Bacillus licheniformis | |
| 4.25 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109E, pH 9.0, 60°C | Bacillus licheniformis | |
| 5.67 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109F, pH 9.0, 60°C | Bacillus licheniformis | |
| 237.5 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109S, pH 9.0, 60°C | Bacillus licheniformis | |
| 266.75 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant wild-type enzyme, pH 9.0, 60°C | Bacillus licheniformis | |
| 435.7 | - |
L-gamma-glutamyl-4-nitroanilide | recombinant mutant R109K, pH 9.0, 60°C | Bacillus licheniformis | |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
