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Literature summary for 3.4.21.62 extracted from

  • Fuchita, N.; Arita, S.; Ikuta, J.; Miura, M.; Shimomura, K.; Motoshima, H.; Watanabe, K.
    Gly or Ala substitutions for Pro210Thr211Asn212 at the beta8-beta9 turn of subtilisin Carlsberg increase the catalytic rate and decrease thermostability (2012), Biochim. Biophys. Acta, 1824, 620-626.
    View publication on PubMed
Search for more literature for 3.4.21.62

Cloned(Commentary)

Cloned (Comment) Organism
cloning in Escherichia coli strain DH5alpha and Saccharomyces cerevisiae strain YPH499, recombinant expression of wild-type and mutant enzymes in Bacillus megaterium strain WH320 from a shuttle expression vector carrying a ribosome-binding site and a signal sequence of extracellular metalloprotease from Bacillus megaterium downstream of xylose isomerase promoter Bacillus licheniformis

Protein Variants

Protein Variants Comment Organism
N212G site-directed mutagenesis, the mutant shows increased kcat compared to the wild-type enzyme Bacillus licheniformis
P210A site-directed mutagenesis, the mutant shows increased kcat compared to the wild-type enzyme Bacillus licheniformis
P210G site-directed mutagenesis, the mutant shows 1.5fold increased kcat compared to the wild-type enzyme Bacillus licheniformis
P210G/T211G site-directed mutagenesis, the mutant shows 1.7fold increased kcat compared to the wild-type enzyme Bacillus licheniformis
P210G/T211G/N212G site-directed mutagenesis, the mutant shows 1.8fold increased kcat compared to the wild-type enzyme Bacillus licheniformis
T211G site-directed mutagenesis, the mutant shows 1.8fold increased kcat compared to the wild-type enzyme Bacillus licheniformis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.227
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant N212G Bacillus licheniformis
0.233
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant wild-type enzyme Bacillus licheniformis
0.278
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant P210G Bacillus licheniformis
0.279
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant wmutant T211G Bacillus licheniformis
0.29
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant P210G/T211G Bacillus licheniformis
0.301
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant P210G/T211G/N212G Bacillus licheniformis
0.325
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant P210A Bacillus licheniformis

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ required for thermostability Bacillus licheniformis

Organism

Organism UniProt Comment Textmining
Bacillus licheniformis P00780
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
N-succinyl-Ala-Ala-Pro-Leu-4-nitroanilide + H2O
-
 Bacillus licheniformis N-succinyl-Ala-Ala-Pro-Leu + 4-nitroaniline
-
 ?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
-
 Bacillus licheniformis N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
 ?
N-succinyl-Ala-Ala-Val-Ala-4-nitroanilide + H2O
-
 Bacillus licheniformis N-succinyl-Ala-Ala-Val-Ala + 4-nitroaniline
-
 ?

Synonyms

Synonyms Comment Organism
subtilisin Carlsberg
-
 Bacillus licheniformis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
60
-
-
 Bacillus licheniformis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
15 65 activity range, profile overview Bacillus licheniformis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
419
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant wild-type enzyme Bacillus licheniformis
462
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant wmutant T211G Bacillus licheniformis
475
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant N212G Bacillus licheniformis
638
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant P210G Bacillus licheniformis
697
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant P210G/T211G Bacillus licheniformis
715
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant P210A Bacillus licheniformis
759
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant P210G/T211G/N212G Bacillus licheniformis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Bacillus licheniformis

General Information

General Information Comment Organism
additional information the catalytic triad residues consists of Ser221, Pro210 to His64 Bacillus licheniformis