Cloned (Comment) | Organism |
---|---|
recombinant expression in Pichia pastoris strain GS115, the enzyme is secreted, the recombinant enzyme level is 7.1 times that of wild-type Bacillus subtilis strain Zj016, subcloning in Escherichia coli strain JM109 | Bacillus subtilis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | strong inhibition | Bacillus subtilis | |
Ca2+ | 68.4% inhibition at 1 mM | Bacillus subtilis | |
Cu2+ | slight inhibition at 1 mM | Bacillus subtilis | |
DTT | complete inhibition at 1 mM, 50°C, 1 h | Bacillus subtilis | |
EDTA | complete inhibition at 1 mM, 50°C, 1 h, the activity can be restored by Co2+ and partially by Zn2+ | Bacillus subtilis | |
Mg2+ | slight inhibition at 1 mM | Bacillus subtilis | |
Mn2+ | slight inhibition at 1 mM | Bacillus subtilis | |
Ni2+ | 92.5% inhibition at 1 mM | Bacillus subtilis | |
PMSF | slight inhibition | Bacillus subtilis | |
Zn2+ | slight inhibition at 1 mM | Bacillus subtilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.97 | - |
Leu-4-nitroanilide | pH 8.5, 50°C, recombinant enzyme | Bacillus subtilis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | activates to 228% activity at 0.1 mM | Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | O32106 | - |
- |
Bacillus subtilis Zj016 | O32106 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | the sequence analysis of aminopeptidase indicates three potential N-glycosylation sites, the N-glycosylation of the aminopeptidase leads to higher thermostability and catalytic efficiency. Secondary structure of recombinant BSAP is analyzed by CD spectroscopy and compared with that of wild-type BSAP revealing that the effect of glycosylation on secondary structure is negligible | Bacillus subtilis |
Purification (Comment) | Organism |
---|---|
recombinant extracellular enzyme 8.4fold from Pichia pastoris strain GS115 culture supernatant by dialysis, anion exchange chromatography, ultrafiltration, and gel filtration | Bacillus subtilis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
132.5 | - |
purified native enzyme, pH 8.5, 50°C, substrate Leu-4-nitroanilide | Bacillus subtilis |
240.48 | - |
purified recombinant enzyme, pH 8.5, 50°C, substrate Leu-4-nitroanilide | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Arg-4-nitroanilide + H2O | 74.5% activity compared to Leu-4-nitroanilide | Bacillus subtilis | Arg + 4-nitroaniline | - |
? | |
Leu-4-nitroanilide + H2O | - |
Bacillus subtilis | Leu + 4-nitroaniline | - |
? | |
Lys-4-nitroanilide + H2O | 20% activity compared to Leu-4-nitroanilide | Bacillus subtilis | Lys + 4-nitroaniline | - |
? | |
Met-4-nitroanilide + H2O | 8.2% activity compared to Leu-4-nitroanilide | Bacillus subtilis | Met + 4-nitroaniline | - |
? | |
additional information | no activity is determined by using His-4-nitroanilide, Ile-4-nitroanilide, Ala-4-nitroanilide, Phe-4-nitroanilide, and Val-4-nitroanilide as substrates | Bacillus subtilis | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BSAP | - |
Bacillus subtilis |
leucine aminopeptidase | - |
Bacillus subtilis |
More | see also EC 3.4.11.10 | Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
recombinant enzyme | Bacillus subtilis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 70 | purified recombinant enzyme, stable at 30-60°C. The enzyme retains 63.5% activity after incubation at 70°C for 1 h, and 75% activity after 3 h at 30-60°C | Bacillus subtilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
recombinant enzyme | Bacillus subtilis |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
8 | 9 | purified recombinant enzyme, the enzyme retains more than 90% activity after incubation at pH 9.0 for 1 h | Bacillus subtilis |