Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Bacillus subtilis by vector pTB51, the enzymes are secreted to the culture medium | Bacillus sp. (in: Bacteria) |
Protein Variants | Comment | Organism |
---|---|---|
L155A | site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is slightly enhanced compared to the wild-type enzyme | Bacillus sp. (in: Bacteria) |
L155A/I156N | site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is enhanced compared to the wild-type enzyme | Bacillus sp. (in: Bacteria) |
L155A/I156V | site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is enhanced compared to the wild-type enzyme | Bacillus sp. (in: Bacteria) |
L155S | site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is slightly enhanced compared to the wild-type enzyme | Bacillus sp. (in: Bacteria) |
L155S/I156N | site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is enhanced compared to the wild-type enzyme | Bacillus sp. (in: Bacteria) |
L155S/I156V | site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is enhanced compared to the wild-type enzyme | Bacillus sp. (in: Bacteria) |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
12 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155S/I156N, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
12 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155S/I156V, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
13 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155A/I156N, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
13 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155A/I156V, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
13 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155S, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
14 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant wild-type enzyme, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
20 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155A, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | TLN contains one zinc ion and four calcium ions, and these ions contribute to enzymatic catalysis and structural stability | Bacillus sp. (in: Bacteria) | |
Zn2+ | TLN contains one zinc ion and four calcium ions, and these ions contribute to enzymatic catalysis and structural stability | Bacillus sp. (in: Bacteria) |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Bacillus sp. (in: Bacteria) | autodegradation at position 154155 | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus sp. (in: Bacteria) | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
proteolytic modification | autodegradation at position 154-155 | Bacillus sp. (in: Bacteria) |
Purification (Comment) | Organism |
---|---|
recombinant extracellular wild-type and mutant enzymes from Bacillus subtilis culture medium by ammonium sulfate fractionation, hydrophobic interaction chromatography, and gel filtration | Bacillus sp. (in: Bacteria) |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | autodegradation at position 154155 | Bacillus sp. (in: Bacteria) | ? | - |
? | |
N-benzyloxycarbonyl-Gly-L-Leu amide + H2O | - |
Bacillus sp. (in: Bacteria) | N-benzyloxycarbonyl-Gly + L-Leu amide | - |
? |
Synonyms | Comment | Organism |
---|---|---|
TLN | - |
Bacillus sp. (in: Bacteria) |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Bacillus sp. (in: Bacteria) |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
half-life of wild-type enzyme is 18.3 min, half-lives of mutants are 25 min for L155A, 24 min for L155S, 60.8 min for L155A/I156V, 62.4 min for L155S/I156N, 93.3 for L155S/I156N, and 40 min for L155S/I156V | Bacillus sp. (in: Bacteria) |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
13.58 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155A/I156V, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
26.67 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155A/I156N, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
48.33 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155S/I156N, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
51.67 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant wild-type enzyme, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
80 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155A, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
105 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155S/I156V, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
165 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155S, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Bacillus sp. (in: Bacteria) |
General Information | Comment | Organism |
---|---|---|
additional information | autodegradation at position 154155 | Bacillus sp. (in: Bacteria) |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.03 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155A/I156V, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
2 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155A/I156N, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
3.67 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant wild-type enzyme, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
4 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155A, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
4 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155S/I156N, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
12 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155S/I156V, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) | |
12.67 | - |
N-benzyloxycarbonyl-Gly-L-Leu amide | recombinant mutant L155S, pH 7.0, 37°C | Bacillus sp. (in: Bacteria) |