Any feedback?
Literature summary extracted from
- Proteolysin, a novel highly thermostable and cosolvent-compatible protease from the thermophilic bacterium Coprothermobacter proteolyticus (2013), Appl. Environ. Microbiol., 79, 5625-5632.
Activating Compound
| EC Number | Activating Compound | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.4.21.66 | cetyltrimethylammonium bromide | CTAB, 5% v/v, activates to 178% activity | Coprothermobacter proteolyticus |  |
| 3.4.21.66 | DTT | - |
Coprothermobacter proteolyticus | |
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.4.21.66 | gene aprE, DNA and amino acid sequence determination and analysis, phylogenetic tree, recombinant expression of wild-type and mutant N-terminally maltose-binding protein-tagged and C-terminally His6-tagged enzymes in Escherichia coli strain C43(DE3) | Coprothermobacter proteolyticus |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.4.21.66 | C182A/C201A | site-directed mutagenesis, elimination of the disulfide bond, the mutant shows increased activity but reduced thermostability compared to the wild-type enzyme | Coprothermobacter proteolyticus |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.4.21.66 | EDTA | 87% inhibition at 10 mM | Coprothermobacter proteolyticus | |
| 3.4.21.66 | guanidinium hydrochloride | 6 M guanidinium hydrochloride gives a 40% reduction of activity after 1 h of incubation at 40°C | Coprothermobacter proteolyticus | |
| 3.4.21.66 | H2O2 | 15% v/v, 90% inhibition | Coprothermobacter proteolyticus | |
| 3.4.21.66 | additional information | the enzyme shows good activity and stability in the presence of organic solvents, detergents, and dithiothreitol, and it remains active in 6 M guanidinium hydrochloride | Coprothermobacter proteolyticus | |
| 3.4.21.66 | PMSF | completely abolishes the activity of proteolysin | Coprothermobacter proteolyticus | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.4.21.66 | 0.33 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C, recombinant mutant C182A/C201A | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 0.41 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C, recombinant wild-type enzyme | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 0.57 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 70°C, recombinant wild-type enzyme | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 0.66 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 70°C, recombinant mutant C182A/C201A | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 1.58 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C, recombinant wild-type enzyme, in presence of DTT | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 1.96 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C, recombinant mutant C182A/C201A, in presence of DTT | Coprothermobacter proteolyticus | |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 3.4.21.66 | extracellular | - |
Coprothermobacter proteolyticus | - |
- |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.4.21.66 | Ca2+ | required, two calcium sites (Ca1 and Ca2) are present in thermitase are likely to be present in proteolysin as well | Coprothermobacter proteolyticus | |
| 3.4.21.66 | additional information | bivalent metal cations increase the activity of proteolysin | Coprothermobacter proteolyticus |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.4.21.66 | Coprothermobacter proteolyticus | B5Y6G0 | gene aprE | - |
Posttranslational Modification
| EC Number | Posttranslational Modification | Comment | Organism |
|---|---|---|---|
| 3.4.21.66 | additional information | recombinant proteolysin processing and maturation requies a heat treatment step. The processing can be performed at 70°C for 12 h or at 80°C for 3 h | Coprothermobacter proteolyticus |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.4.21.66 | recombinant wild-type and mutant N-terminally maltose-binding protein-tagged and C-terminally His6-tagged enzymes from Escherichia coli strain C43(DE3) to near homogeneity from crude cell lysate by heat treatment at 80°C for 3 h, and ultrafiltration | Coprothermobacter proteolyticus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.4.21.66 | azocasein + H2O | proteinolytic activity | Coprothermobacter proteolyticus | ? | - |
? | |
| 3.4.21.66 | additional information | proteolytic activities of cell extracts and purified enzyme samples are monitored on 5% skim milk-LB agar plates or by azocasein assey | Coprothermobacter proteolyticus | ? | - |
? | |
| 3.4.21.66 | N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O | amidolytic activity | Coprothermobacter proteolyticus | N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline | - |
? | |
| 3.4.21.66 | oxidized insulin B chain + H2O | peptidolytic activity, primary cleavage site of proteolysin is located between Ala14 and Leu15. Prolonged incubation of 24 h reveals additional cleavage sites after Phe1, Asn3, Gln4, Leu17, Cys19, Phe24, Phe25, Tyr25, and Lys29, which shows that proteolysin has a relaxed cleavage site specificity | Coprothermobacter proteolyticus | ? | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.4.21.66 | PrlA | - |
Coprothermobacter proteolyticus |
| 3.4.21.66 | proteolysin | - |
Coprothermobacter proteolyticus |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.4.21.66 | 40 | 70 | substrate N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | Coprothermobacter proteolyticus |
| 3.4.21.66 | 85 | - |
substrate azocasein | Coprothermobacter proteolyticus |
Temperature Range [°C]
| EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.4.21.66 | 20 | 80 | activity range, recombinant enzyme | Coprothermobacter proteolyticus |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.4.21.66 | 70 | - |
purified recombinant enzyme, substrate N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide, 20 h, 35% activity remaining | Coprothermobacter proteolyticus |
| 3.4.21.66 | 80 | 100 | thermostability profiles of the purified recombinant enzyme with different substrates, overview | Coprothermobacter proteolyticus |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.4.21.66 | 25 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C, recombinant wild-type enzyme | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 37 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C, recombinant wild-type enzyme, in presence of DTT | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 110 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 70°C, recombinant wild-type enzyme | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 111 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C, recombinant mutant C182A/C201A | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 124 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C, recombinant mutant C182A/C201A, in presence of DTT | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 248 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 70°C, recombinant mutant C182A/C201A | Coprothermobacter proteolyticus | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.4.21.66 | 8 | - |
- |
Coprothermobacter proteolyticus |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.4.21.66 | 7 | 9 | activity range | Coprothermobacter proteolyticus |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 3.4.21.66 | evolution | proteolysin is a serine protease of the thermitase subgroup of subtilases, phylogenetic tree | Coprothermobacter proteolyticus |
| 3.4.21.66 | additional information | structure homology modelling, overview | Coprothermobacter proteolyticus |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.4.21.66 | 62 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C, recombinant wild-type enzyme | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 65 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 70°C, recombinant wild-type enzyme | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 69 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C, recombinant wild-type enzyme, in presence of DTT | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 126 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C, recombinant mutant C182A/C201A, in presence of DTT | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 189 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 70°C, recombinant mutant C182A/C201A | Coprothermobacter proteolyticus | |
| 3.4.21.66 | 335 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C, recombinant mutant C182A/C201A | Coprothermobacter proteolyticus | |
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
