Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Picrophilus torridus |
Protein Variants | Comment | Organism |
---|---|---|
H369N | mutation introduces significant transpeptidase activity. Mutation results in lowering of KM in hydrolysis of gamma-L-glutamyl-4-nitroanilide and in decline in hydrolytic rate | Picrophilus torridus |
R346A | mutation results in lowering of KM in hydrolysis of gamma-L-glutamyl-4-nitroanilide and in decline in hydrolytic rate | Picrophilus torridus |
R346E | mutation results in lowering of KM in hydrolysis of gamma-L-glutamyl-4-nitroanilide and in decline in hydrolytic rate | Picrophilus torridus |
Y327E | mutation results in lowering of KM in hydrolysis of gamma-L-glutamyl-4-nitroanilide and in decline in hydrolytic rate | Picrophilus torridus |
Y327N | mutation introduces significant transpeptidase activity. Mutation results in lowering of KM in hydrolysis of gamma-L-glutamyl-4-nitroanilide and in decline in hydrolytic rate. The mutant enzyme retains more than 90% transpeptidase activity in the presence of Ba2+, Ca2+, Cu2+, Mg2+, Mn2+, and Zn2+ and is completely inhibited by Cd2+, Co2+, Fe2+, Hg2+, Ni2+ and Pb2+ | Picrophilus torridus |
Y349D | mutation results in lowering of KM in hydrolysis of gamma-L-glutamyl-4-nitroanilide and in decline in hydrolytic rate | Picrophilus torridus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
6-diazo-4 oxonorleucine | 1 mM, complete inhibition of hydrolysis of gamma-L-glutamyl-4-nitroanilide | Picrophilus torridus | |
azaserine | 1 mM, complete inhibition of hydrolysis of gamma-L-glutamyl-4-nitroanilide | Picrophilus torridus | |
Cu2+ | - |
Picrophilus torridus | |
additional information | the enzyme retains more than 90% of the gamma-L-glutamyl-4-nitroanilide hydrolase activity in the presence of most of divalent cations like Ba2+, Ca2+, Co2+, Cd2+, Fe2+, Hg2+, Mg2+, Mn2+ and Zn2+. The enzyme retains more than 90% of the gamma-L-glutamyl-4-nitroanilide hydrolase activity in the presence of 10 mM chelating agents like EDTA, EGTA and 1,10-phenanthroline and is not inhibited by N-bromosuccinimide and iodoacetic acid. Reducing agents like dithiothreitol and 2-mercaptoethanol have no significant effect on the activity | Picrophilus torridus | |
Ni2+ | - |
Picrophilus torridus | |
phenylmethylsulfonyl fluoride | 1 mM, complete inhibition of hydrolysis of gamma-L-glutamyl-4-nitroanilide | Picrophilus torridus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0143 | - |
gamma-L-glutamyl-4-nitroanilide | pH 7.0, temperature not specified in the publication, hydrolytic activity, mutant enzyme H368S | Picrophilus torridus | |
0.025 | - |
gamma-L-glutamyl-4-nitroanilide | pH 7.0, temperature not specified in the publication, transpeptidase activity with Gly-Gly as acceptor, mutant enzyme H368S | Picrophilus torridus | |
0.025 | - |
gamma-L-glutamyl-4-nitroanilide | pH 7.0, temperature not specified in the publication, transpeptidase activity with Gly-Gly as acceptor, mutant enzyme Y327N | Picrophilus torridus | |
0.067 | - |
gamma-L-glutamyl-4-nitroanilide | pH 7.0, temperature not specified in the publication, hydrolytic activity, mutant enzyme Y327N | Picrophilus torridus | |
0.1 | - |
gamma-L-glutamyl-4-nitroanilide | pH 7.0, temperature not specified in the publication, hydrolytic activity, wild-type enzyme | Picrophilus torridus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
17000 | - |
1 * 17000 + 1 * 30000, SDS-PAGE | Picrophilus torridus |
30000 | - |
1 * 17000 + 1 * 30000, SDS-PAGE | Picrophilus torridus |
47000 | - |
unprocessed precursor, SDS-PAGE | Picrophilus torridus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Picrophilus torridus | Q6KZT2 | - |
- |
Picrophilus torridus DSM 9790 | Q6KZT2 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Picrophilus torridus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
gamma-L-glutamyl-4-nitroanilide + Gly-Gly | the enzyme also catalyzes the hydrolase reaction: gamma-L-glutamyl-4-nitroanilide + H2O = L-glutamate + 4-nitroaniline | Picrophilus torridus | 4-nitroaniline + gamma-L-glutamyl-Gly-Gly | - |
? | |
gamma-L-glutamyl-4-nitroanilide + Gly-Gly | the enzyme also catalyzes the hydrolase reaction: gamma-L-glutamyl-4-nitroanilide + H2O = L-glutamate + 4-nitroaniline | Picrophilus torridus DSM 9790 | 4-nitroaniline + gamma-L-glutamyl-Gly-Gly | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterodimer | 1 * 17000 + 1 * 30000, SDS-PAGE | Picrophilus torridus |
Synonyms | Comment | Organism |
---|---|---|
gamma-glutamyl transpeptidase | - |
Picrophilus torridus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
hydrolysis of gamma-L-glutamyl-4-nitroanilide | Picrophilus torridus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 60 | more than 40% of maximal activity, hydrolysis of gamma-L-glutamyl-4-nitroanilide | Picrophilus torridus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
24 h, no loss of gamma-L-glutamyl-4-nitroanilide hydrolase activity | Picrophilus torridus |
50 | - |
t1/2: 1 h, gamma-L-glutamyl-4-nitroanilide hydrolase activity | Picrophilus torridus |
60 | - |
t1/2: 30 min, gamma-L-glutamyl-4-nitroanilide hydrolase activity | Picrophilus torridus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
hydrolysis of gamma-L-glutamyl-4-nitroanilide | Picrophilus torridus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 9 | more than 40% of maximal activity, hydrolysis of gamma-glutamyl-4-nitroanilide | Picrophilus torridus |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
3 | 10 | 25°C, 1 h, more than 50% residual activity | Picrophilus torridus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
25 | - |
gamma-L-glutamyl-4-nitroanilide | pH 7.0, temperature not specified in the publication, hydrolytic activity, mutant enzyme Y327N | Picrophilus torridus | |
39.3 | - |
gamma-L-glutamyl-4-nitroanilide | pH 7.0, temperature not specified in the publication, hydrolytic activity, wild-type enzyme | Picrophilus torridus | |
101 | - |
gamma-L-glutamyl-4-nitroanilide | pH 7.0, temperature not specified in the publication, hydrolytic activity, mutant enzyme H368S | Picrophilus torridus | |
172.8 | - |
gamma-L-glutamyl-4-nitroanilide | pH 7.0, temperature not specified in the publication, transpeptidase activity with Gly-Gly as acceptor, mutant enzyme H368S | Picrophilus torridus | |
182.3 | - |
gamma-L-glutamyl-4-nitroanilide | pH 7.0, temperature not specified in the publication, transpeptidase activity with Gly-Gly as acceptor, mutant enzyme Y327N | Picrophilus torridus |