Cloned (Comment) | Organism |
---|---|
gene aprE, construction of error-prone PCR libraries, recombinant expression of wild-type and mutant enzymes in Bacillus subtilis strain WB600 and secretion to the culture medium | Bacillus subtilis |
Protein Variants | Comment | Organism |
---|---|---|
A153V | site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, but does not show improved properties in chaotropic conditions | Bacillus subtilis |
G166M | site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, and improved properties in chaotropic conditions | Bacillus subtilis |
G166S | site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, and improved properties in chaotropic conditions | Bacillus subtilis |
I205V | site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, but does not show improved properties in chaotropic conditions | Bacillus subtilis |
additional information | directed evolution of subtilisin E into a highly active and guanidinium chloride- and sodium dodecylsulfate-tolerant protease, subtilisin E is engineered into a chaotolerent protease, the enzyme gains a protease stability improved in a chaotropic salt (GdmCl) or a detergent (SDS), nultiple screening, overview | Bacillus subtilis |
S62I | site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, and slightly improved properties in chaotropic conditions | Bacillus subtilis |
S62I/A153V/G166S/I205V | site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, and highly improved properties in chaotropic conditions | Bacillus subtilis |
S62I/G166M | site-directed mutagenesis, the mutant shows increased activity without detergents compared to the wild-type, and highly improved properties in chaotropic conditions | Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | P04189 | gene aprE | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Bacillus subtilis strain WB600 culture supernatant by anion and cation exchange chromatography and ultrafilatration | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzyme shows activity towards complex substrates, e.g. skimmed milk | Bacillus subtilis | ? | - |
? | |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide + H2O | - |
Bacillus subtilis | succinyl-L-Ala-L-Ala-L-Pro-L-Phe + 7-amino-4-methylcoumarin | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Subtilisin E | - |
Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Bacillus subtilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
15 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant wild-type enzyme, in presence of 2 M GdmCl | Bacillus subtilis | |
34 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant mutant S62I/A153V/G166S/I205V, in presence of 2 M GdmCl | Bacillus subtilis | |
35 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant wild-type enzyme, in presence of SDS | Bacillus subtilis | |
42 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant mutant S62I/A153V/G166S/I205V, in absence of GdmCl or SDS | Bacillus subtilis | |
45 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant mutant S62I/A153V/G166S/I205V, in presence of SDS | Bacillus subtilis | |
50 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant wild-type enzyme, in absence of GdmCl or SDS | Bacillus subtilis | |
62 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant mutant G166M, in presence of 2 M GdmCl | Bacillus subtilis | |
65 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant mutant S62I, in presence of SDS | Bacillus subtilis | |
70 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant mutant S62I/G166M, in presence of 2 M GdmCl | Bacillus subtilis | |
81 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant mutant S62I/G166M, in presence of SDS | Bacillus subtilis | |
102 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant mutant S62I, in absence of GdmCl or SDS | Bacillus subtilis | |
104 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant mutant G166M, in presence of SDS | Bacillus subtilis | |
123 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant mutant S62I/G166M, in absence of GdmCl or SDS | Bacillus subtilis | |
141 | - |
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-methyl-coumaryl-7-amide | pH 8.5, 37°C, recombinant mutant G166M, in absence of GdmCl or SDS | Bacillus subtilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Bacillus subtilis |
General Information | Comment | Organism |
---|---|---|
additional information | structure molecular modeling using the crystal structure of the pro-subtilisin E complex, PDB ID 1SCJ, the active site Cys221 is replaced with the catalytic serine, molecular dynamics simulation, overview | Bacillus subtilis |