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Literature summary for 3.4.24.B2 extracted from
- Characterization of the novel thermostable oligopeptidase from Geobacillus thermoleovorans DSM 15325 (2015), J. Microbiol. Biotechnol., 25, 1070-1083.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| glycerol | 26% inhibition at 5%, 15% ativation at 1% | Geobacillus thermoleovorans |  |
| TCEP | 10% activation at 1 mM | Geobacillus thermoleovorans | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene GT-SM3B, DNA and amino acid sequence determination and analysis, sequence comparisons, expression of the soluble enzyme with introduced N-terminal hexahistidine thrombin-cleavable tag in Escherichia coli strain BL21 (DE3) cytoplasm and secretion of the enzyme | Geobacillus thermoleovorans |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | 14% inhibition at 1 mM | Geobacillus thermoleovorans | |
| Brij 35 | complete inhibition at 1 mM | Geobacillus thermoleovorans | |
| Brij 58 | complete inhibition at 1 mM | Geobacillus thermoleovorans | |
| Co2+ | 6% inhibition at 1 mM | Geobacillus thermoleovorans | |
| Cu2+ | 47% inhibition at 1 mM | Geobacillus thermoleovorans | |
| DTT | 42% inhibition at 1 mM | Geobacillus thermoleovorans | |
| ethylene glycol | 34% inhibition at 1% | Geobacillus thermoleovorans | |
| Fe3+ | 77% inhibition at 1 mM | Geobacillus thermoleovorans | |
| glycerol | 26% inhibition at 5%, 15% ativation at 1% | Geobacillus thermoleovorans | |
| Guanidine-HCl | 22% inhibition at 1 mM | Geobacillus thermoleovorans | |
| Mn2+ | 51% inhibition at 1 mM | Geobacillus thermoleovorans | |
| Ni2+ | 41% inhibition at 1 mM | Geobacillus thermoleovorans | |
| SDS | 48% inhibition at 1 mM, complete inhibition at 10 mM | Geobacillus thermoleovorans | |
| Triton X-100 | 37% inhibition at 1% | Geobacillus thermoleovorans | |
| Triton X-305 | 98% inhibition at 1% | Geobacillus thermoleovorans | |
| Tween 20 | 33% inhibition at 0.1% | Geobacillus thermoleovorans | |
| Tween 40 | 42% inhibition at 0.1% | Geobacillus thermoleovorans | |
| Tween 60 | 38% inhibition at 0.1% | Geobacillus thermoleovorans | |
| Tween 80 | 58% inhibition at 0.1% | Geobacillus thermoleovorans | |
| Zn2+ | 14% inhibition at 1 mM | Geobacillus thermoleovorans | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00217 | - |
Carbobenzoxy-Gly-Pro-Gly-Gly-Pro-Ala | pH 7.3, 40°C, recombinant wild-type enzyme | Geobacillus thermoleovorans | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| extracellular | the enzyme sequence contains a 23-residue signal peptide | Geobacillus thermoleovorans | - |
- |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Li+ | 5% activation at 1 mM | Geobacillus thermoleovorans | |
| Mg2+ | 21% activation at 1 mM | Geobacillus thermoleovorans | |
| additional information | no effect by 1 mM K+ and 1-5 mM Ca2+ | Geobacillus thermoleovorans | |
| Na+ | 27% activation at 1 mM | Geobacillus thermoleovorans | |
| Zn2+ | the enzyme contains a conservative zinc metallopeptidase motif, His400-Glu401-X-XHis404 | Geobacillus thermoleovorans | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 67700 | - |
2 * 67700, enzyme without signal peptide, SDS-PAGE, 2 * 70200, enzyme with signal peptide, SDS-PAGE | Geobacillus thermoleovorans |
| 70200 | - |
2 * 67700, enzyme without signal peptide, SDS-PAGE, 2 * 70200, enzyme with signal peptide, SDS-PAGE | Geobacillus thermoleovorans |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Geobacillus thermoleovorans | W5RWH8 | formerly Geobacillus lituanicus DSM 15325, gene GT-SM3B | - |
| Geobacillus thermoleovorans DSM 15325 | W5RWH8 | formerly Geobacillus lituanicus DSM 15325, gene GT-SM3B | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant secreted enzyme from Escherichia coli strain BL21 (DE3) culture medium by dialysis, anion exchange and cation exchange chromatography | Geobacillus thermoleovorans |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| carbobenzoxy-Gly-Pro-Gly-Gly-Pro-Ala + H2O | - |
Geobacillus thermoleovorans | ? | - |
? | |
| carbobenzoxy-Gly-Pro-Gly-Gly-Pro-Ala + H2O | - |
Geobacillus thermoleovorans DSM 15325 | ? | - |
? | |
| additional information | the enzyme shows hydrolytic activity against short-chain collagenous peptides, gelatin-derived oligopeptides, type I collagen-derived oligopeptides, and raw collagen extract oligopeptides | Geobacillus thermoleovorans | ? | - |
? | |
| additional information | the enzyme shows hydrolytic activity against short-chain collagenous peptides, gelatin-derived oligopeptides, type I collagen-derived oligopeptides, and raw collagen extract oligopeptides | Geobacillus thermoleovorans DSM 15325 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homodimer | 2 * 67700, enzyme without signal peptide, SDS-PAGE, 2 * 70200, enzyme with signal peptide, SDS-PAGE | Geobacillus thermoleovorans |
| More | both the homodimer and monomer are catalytically active | Geobacillus thermoleovorans |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| GT-SM3B | - |
Geobacillus thermoleovorans |
| oligopeptidase F | UniProt | Geobacillus thermoleovorans |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
- |
Geobacillus thermoleovorans |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 10 | 75 | activity range, profile overview | Geobacillus thermoleovorans |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 50 | - |
purified recombinant enzyme, 84% activity remaining | Geobacillus thermoleovorans |
| 60 | - |
purified recombinant enzyme, 71% activity remaining after 1 h | Geobacillus thermoleovorans |
| 70 | - |
purified recombinant enzyme, 30% activity remaining after 1 h | Geobacillus thermoleovorans |
| 80 | - |
purified recombinant enzyme, no activity remaining after 1 h | Geobacillus thermoleovorans |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 5.99 | - |
Carbobenzoxy-Gly-Pro-Gly-Gly-Pro-Ala | pH 7.3, 40°C, recombinant wild-type enzyme | Geobacillus thermoleovorans | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.3 | - |
- |
Geobacillus thermoleovorans |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 4 | 8.5 | activity range, profile overview | Geobacillus thermoleovorans |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 5 | 10 | recombinant enzyme, stable at with over 60% of maximal activity | Geobacillus thermoleovorans |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme belongs to the M3B subfamily of metallopeptidases and displays the highest amino acid sequence identity (40.3%) to the oligopeptidase PepFBa from mesophilic Bacillus amyloliquefaciens 23-7A among the characterized oligopeptidases | Geobacillus thermoleovorans |
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