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Literature summary for 3.4.21.62 extracted from

  • Li, Z.; Roccatano, D.; Lorenz, M.; Schwaneberg, U.
    Directed evolution of subtilisin E into a highly active and guanidinium chloride- and sodium dodecylsulfate-tolerant protease (2012), ChemBioChem, 13, 691-699.
    View publication on PubMed
Search for more literature for 3.4.21.62

Cloned(Commentary)

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

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

Organism UniProt Comment Textmining
Bacillus subtilis P04189 gene aprE
-

Purification (Commentary)

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 and Products (Substrate)

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

Synonyms Comment Organism
Subtilisin E
-
 Bacillus subtilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Bacillus subtilis

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
 assay at Bacillus subtilis

General Information

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