Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination, subcloning and expression of the recombinant enzyme possessing a Asp70 and Asp184 residues in Escherichia coli strain BL21(DE3), the soluble enzyme is inducible by growth on 1 M sorbitol, optimization of the expression method | Streptomyces griseus |
Protein Variants | Comment | Organism |
---|---|---|
E131 | site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type enzyme | Streptomyces griseus |
Y246 | site-directed mutagenesis, the mutant enzyme shows highly reduced activity compared to the wild-type enzyme | Streptomyces griseus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | - |
Streptomyces griseus | |
Zn2+ | the enzyme is a double-zinc aminopeptidase | Streptomyces griseus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces griseus | - |
DSM 40855 | - |
Purification (Comment) | Organism |
---|---|
recombinant soluble wild-type and mutant enzymes 28fold from Escherichia coli strain BL21(DE3) by heat treatment for 20 min at 50°C and anion exchange chromatography | Streptomyces griseus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues | catalytic pathway and reaction mechanism, catalytic residues are Glu131 and Tyr246, Tyr246 is involved in stabilization of the reaction transition state intermediate, also residue Glu151 is involved | Streptomyces griseus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
830 | - |
purified recombinant wild-type enzyme | Streptomyces griseus |
Synonyms | Comment | Organism |
---|---|---|
double-zinc aminopeptidase | - |
Streptomyces griseus |
SGAP | - |
Streptomyces griseus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Streptomyces griseus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
20 min, stable | Streptomyces griseus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Streptomyces griseus |