Cloned (Comment) | Organism |
---|---|
recombinant enzyme expression from expression vector pTONA5a, which includes a promoter from Streptomyces metalloendopeptidase, in Streptomyces lividans strain 1326 | Streptomyces griseus |
recombinant enzyme expression from expression vector pTONA5a, which includes a promoter from Streptomyces metalloendopeptidase, in Streptomyces lividans strain 1326 | Streptomyces omiyaensis |
Protein Variants | Comment | Organism |
---|---|---|
L71R/S172A | site-directed mutagenesis, inactive active site mutant | Streptomyces griseus |
L71Y | site-directed mutagenesis, the mutant shows a change in topological specificity and high hydrolytic activity toward type IV collagen. The active site mutant shows slightly lower activities toward type I collagen than the wild-type | Streptomyces griseus |
L71Y/Q72R | site-directed mutagenesis, the mutant shows a change in topological specificity and high hydrolytic activity toward type IV collagen. The active site mutant shows slightly lower activities toward type I collagen than the wild-type | Streptomyces griseus |
Q72R | site-directed mutagenesis, the mutant shows a change in topological specificity and high hydrolytic activity toward type IV collagen. The mutant shows slightly lower activities toward type I collagen than the wild-type | Streptomyces griseus |
S172A | site-directed mutagenesis, inactive active site mutant | Streptomyces griseus |
S172A | site-directed mutagenesis, inactive active site mutant | Streptomyces omiyaensis |
Y71L | site-directed mutagenesis, the mutant displays high activity toward type I collagen similar to the wild-type enzyme | Streptomyces omiyaensis |
Y71L/S172A | site-directed mutagenesis, inactive active site mutant | Streptomyces omiyaensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0061 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | wild-type enzyme, pH 8.0, 37°C | Streptomyces omiyaensis | |
0.012 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | mutant L71Y/Q72R, pH 8.0, 37°C | Streptomyces griseus | |
0.0172 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | mutant L71Y, pH 8.0, 37°C | Streptomyces griseus | |
0.0175 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | mutant Y71L, pH 8.0, 37°C | Streptomyces omiyaensis | |
0.0177 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | wild-type enzyme, pH 8.0, 37°C | Streptomyces griseus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | required | Streptomyces griseus | |
Ca2+ | required | Streptomyces omiyaensis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces griseus | - |
- |
- |
Streptomyces omiyaensis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Streptomyces lividans strain 1326 culture supernatant by cation exchange chromatography and dialysis | Streptomyces griseus |
recombinant enzyme from Streptomyces lividans strain 1326 culture supernatant by cation exchange chromatography and diialysis | Streptomyces omiyaensis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
66.1 | - |
wild-type enzyme, pH 8.0, 37°C, substrate benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | Streptomyces griseus |
90.3 | - |
mutant L71Y, pH 8.0, 37°C, substrate benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | Streptomyces griseus |
110.2 | - |
mutant L71Y/Q72R, pH 8.0, 37°C, substrate benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | Streptomyces griseus |
190.2 | - |
mutant Y71L, pH 8.0, 37°C, substrate benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | Streptomyces omiyaensis |
501.5 | - |
wild-type enzyme, pH 8.0, 37°C, substrate benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | Streptomyces omiyaensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide + H2O | - |
Streptomyces griseus | benzyloxycarbonylglycyl-L-prolyl-L-arginine + 7-amino-4-methylcoumarin | - |
? | |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide + H2O | - |
Streptomyces omiyaensis | benzyloxycarbonylglycyl-L-prolyl-L-arginine + 7-amino-4-methylcoumarin | - |
? | |
bovine skin DQ-collagen type I + H2O | - |
Streptomyces griseus | ? | - |
? | |
bovine skin DQ-collagen type I + H2O | - |
Streptomyces omiyaensis | ? | - |
? | |
human placenta DQ-collagen type IV + H2O | - |
Streptomyces griseus | ? | - |
? | |
human placenta DQ-collagen type IV + H2O | - |
Streptomyces omiyaensis | ? | - |
? | |
additional information | three-dimensional structure and topological specificities, substrate recognition in which residue 71 plays a crucial role, overview. Identification of amino acid residue(s) related to collagen binding of wild-type and mutant enzymes | Streptomyces omiyaensis | ? | - |
? | |
additional information | three-dimensional structure and topological specificities, substrate recognition, overview. The neighboring residues Leu71 and Gln72 in the N-terminal beta-barrel domain of the enzyme synergistically play an important role in substrate recognition. Identification of amino acid residue(s) related to collagen binding for wild-type and mutant enzymes | Streptomyces griseus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | three-dimensional structure and topological specificities, substrate recognition, overview | Streptomyces griseus |
More | three-dimensional structure and topological specificities, substrate recognition, overview | Streptomyces omiyaensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Streptomyces griseus |
37 | - |
assay at | Streptomyces omiyaensis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
33 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | wild-type enzyme, pH 8.0, 37°C | Streptomyces griseus | |
51.3 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | mutant L71Y, pH 8.0, 37°C | Streptomyces griseus | |
57.3 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | mutant L71Y/Q72R, pH 8.0, 37°C | Streptomyces griseus | |
93 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | mutant Y71L, pH 8.0, 37°C | Streptomyces omiyaensis | |
279.5 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | wild-type enzyme, pH 8.0, 37°C | Streptomyces omiyaensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Streptomyces griseus |
8 | - |
assay at | Streptomyces omiyaensis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0019 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | wild-type enzyme, pH 8.0, 37°C | Streptomyces griseus | |
0.0032 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | mutant L71Y, pH 8.0, 37°C | Streptomyces griseus | |
0.0048 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | mutant L71Y/Q72R, pH 8.0, 37°C | Streptomyces griseus | |
0.0054 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | mutant Y71L, pH 8.0, 37°C | Streptomyces omiyaensis | |
0.0419 | - |
benzyloxycarbonylglycyl-L-prolyl-L-arginine-4-methylcoumaryl-7-amide | wild-type enzyme, pH 8.0, 37°C | Streptomyces omiyaensis |