Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.4.21.62 extracted from

  • Despotovic, D.; Vojcic, L.; Blanusa, M.; Maurer, K.H.; Zacharias, M.; Bocola, M.; Martinez, R.; Schwaneberg, U.
    Redirecting catalysis from proteolysis to perhydrolysis in subtilisin Carlsberg (2013), J. Biotechnol., 167, 279-286.
    View publication on PubMed
Search for more literature for 3.4.21.62

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of mutant enzyme sin Bacillus subtilis strain DB104, subcloning in Escherichia coli strain DH5alpha Bacillus licheniformis

Protein Variants

Protein Variants Comment Organism
G165X site-directed mutagenesis, catalytic constants of subtilisin Carlsberg for perhydrolysis of methylpropionate, methylbutyrate and methylpentanoate are increased up to 3.5fold, 5.4fold and 5.5fold, respectively, while proteolysis is decreased up to 100fold for N-succinyl-Ala-Ala-Pro-Phe-pnitroanilide substrate Bacillus licheniformis
additional information site saturation mutagenesis of the subtilisin Carlsberg variant T58A/L216W at positionsThr33, Val67, Ser124, Gly126, Ser155, Gly165, Ala168, Trp216, Asn217, Gly218, Met221 Bacillus licheniformis
T58A/L216W site-directed mutagenesis, the mutant catalyzes in addition to its proteolytic activity the generation of peroxycarboxylic acids from corresponding esters in the presence of hydrogen peroxide Bacillus licheniformis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetic analysis of the perhydrolytic wild-type and mutant enzyme activities, overview Bacillus licheniformis
0.15
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/L216W, pH 7.5, 30°C Bacillus licheniformis
0.2
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/G165Y/L216W, pH 7.5, 30°C Bacillus licheniformis
0.42
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/G165L/L216W, pH 7.5, 30°C Bacillus licheniformis
0.54
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/G165I/L216W, pH 7.5, 30°C Bacillus licheniformis
0.59
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant wild-type enzyme, pH 7.5, 30°C Bacillus licheniformis

Organism

Organism UniProt Comment Textmining
Bacillus licheniformis P00780
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information besides ester hydrolysis the enzyme also performs ester perhydrolysis reacting an ester and a hydroperpxide, residue Gl165 in the S1 binding pocket of the enzyme is involved. Perhydrolysis reaction analysis and kinetcis, overview Bacillus licheniformis ?
-
 ?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
-
 Bacillus licheniformis N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
 ?

Synonyms

Synonyms Comment Organism
subtilisin Carlsberg
-
 Bacillus licheniformis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Bacillus licheniformis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.7
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/G165I/L216W, pH 7.5, 30°C Bacillus licheniformis
15.1
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/G165L/L216W, pH 7.5, 30°C Bacillus licheniformis
22.8
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/G165Y/L216W, pH 7.5, 30°C Bacillus licheniformis
33.1
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/L216W, pH 7.5, 30°C Bacillus licheniformis
172.6
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant wild-type enzyme, pH 7.5, 30°C Bacillus licheniformis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Bacillus licheniformis

General Information

General Information Comment Organism
additional information molecular modeling and docking analysis using crystal structure PDB ID 1yu6 Bacillus licheniformis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/G165I/L216W, pH 7.5, 30°C Bacillus licheniformis
36
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/G165L/L216W, pH 7.5, 30°C Bacillus licheniformis
114
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/G165Y/L216W, pH 7.5, 30°C Bacillus licheniformis
220.7
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/L216W, pH 7.5, 30°C Bacillus licheniformis
292.5
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant wild-type enzyme, pH 7.5, 30°C Bacillus licheniformis