Cloned (Comment) | Organism |
---|---|
expressed in Pichia pastoris GS115 cells | Rhizomucor pusillus |
Protein Variants | Comment | Organism |
---|---|---|
A101T | the proteolytic activity of the mutant is similar to the non-mutant enzyme. The milk clotting activity of the mutant is increased compared to the non-mutant enzyme | Rhizomucor pusillus |
A101T/G186D | the proteolytic activity of the mutant is similar to the non-mutant enzyme. The milk clotting activity of the mutant is increased compared to the non-mutant enzyme | Rhizomucor pusillus |
E13A | the proteolytic activity of the mutant decreases remarkably to almost a half of that of non-mutant enzyme | Rhizomucor pusillus |
E13D | the proteolytic activity of the mutant decreases remarkably to almost a half of that of non-mutant enzyme | Rhizomucor pusillus |
E13P | the proteolytic activity of the mutant decreases remarkably to almost a half of that of non-mutant enzyme | Rhizomucor pusillus |
E13Q | the proteolytic activity of the mutant decreases remarkably to almost a half of that of non-mutant enzyme | Rhizomucor pusillus |
G186D | the proteolytic activity of the mutant is similar to the non-mutant enzyme. The milk clotting activity of the mutant is increased compared to the non-mutant enzyme | Rhizomucor pusillus |
G186D/E13D | the mutant shows a significant milk-clotting activity. The mutant rennet can decrease hydrolysis of protein during ripening of cheese. The mutant has a reduction in thermostability along with a sharp decrease in proteolytic activity | Rhizomucor pusillus |
G186D/E13Q | the mutant has a reduction in thermostability along with a sharp decrease in proteolytic activity | Rhizomucor pusillus |
G186D/E13Q | the mutant shows a significant milk-clotting activity. The mutant rennet can decrease hydrolysis of protein during ripening of cheese. The mutant has a reduction in thermostability along with a sharp decrease in proteolytic activity | Rhizomucor pusillus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1,10-phenanthroline | complete inhibition at 1 mM | Rhizomucor pusillus | |
Aprotinin | 72.6% residual activity at 1 mM | Rhizomucor pusillus | |
EDTA | 71.1% residual activity at 50 mM | Rhizomucor pusillus | |
Fe2+ | 87.7% residual activity at 5 mM | Rhizomucor pusillus | |
Fe3+ | 85.8% residual activity at 5 mM | Rhizomucor pusillus | |
leupeptin | 96.5% residual activity at 0.2 mM | Rhizomucor pusillus | |
additional information | not inhibited by Cu2+ and Na+ | Rhizomucor pusillus | |
Ni2+ | 47.8% residual activity at 5 mM | Rhizomucor pusillus | |
pepstatin A | 24.5% residual activity at 0.05 mM | Rhizomucor pusillus | |
phenylmethylsulfonyl fluoride | 93.1% residual activity at 1 mM | Rhizomucor pusillus | |
Zn2+ | 85.5% residual activity at 5 mM | Rhizomucor pusillus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | 218.8% activity at 5 mM | Rhizomucor pusillus | |
K+ | 117.3% activity at 5 mM | Rhizomucor pusillus | |
Mg2+ | 106.8% activity at 5 mM | Rhizomucor pusillus | |
Mn2+ | 120.7% activity at 5 mM | Rhizomucor pusillus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhizomucor pusillus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
ammonium sulfate precipitation, DEAE-52 column chromatography and Sephadex G-100 gel filtration | Rhizomucor pusillus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
casein + H2O | - |
Rhizomucor pusillus | ? | - |
? | |
skimmed milk + H2O | - |
Rhizomucor pusillus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 44000, SDS-PAGE | Rhizomucor pusillus |
Synonyms | Comment | Organism |
---|---|---|
Aspartic proteinase | - |
Rhizomucor pusillus |
MPR | - |
Rhizomucor pusillus |
Mucor rennin | - |
Rhizomucor pusillus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
purified mutant G186D/E13D and non-mutant enzymes act optimally at 60°C and later, they start losing their activity rapidly leading to complete inactivation at 75°C | Rhizomucor pusillus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
the wild type enzyme retains 88.6% activity after 60 min at 55°C. Mutant enzyme G186D/E13D loses 30% of its activity after 50 min at 55°C | Rhizomucor pusillus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
- |
Rhizomucor pusillus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 7 | about 85% activity at pH 5.0, 100% activity at pH 5.5, about 90% activity at pH 6.0, about 65% activity at pH 6.5, about 40% activity at pH 7.0 | Rhizomucor pusillus |