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Literature summary for 3.4.23.1 extracted from

  • Sinkovits, A.F.; Bryksa, B.C.; Tanaka, T.; Yada, R.Y.
    Understanding the structure-function role of specific catalytic residues in a model food related enzyme: Pepsin (2007), Enzyme Microb. Technol., 40, 1175-1180.
No PubMed abstract available
Search for more literature for 3.4.23.1

Cloned(Commentary)

Cloned (Comment) Organism
wild-type, D32E, D215E, and D32E/D215E thioredoxin-pepsinogen fusion proteins are expressed in Escherichia coli GI724 Sus scrofa

Protein Variants

Protein Variants Comment Organism
D215E unlike wild-type, the mutant fusion protein is incapable of autocatalytic activation upon acidification of the medium, as determined by Western blot analysis. Mature mutant pepsin is obtained by processing fusion protein samples through an immobilized pepsin column. Tm-value of mutant enzyme is 65°C compared to 71°C for wild-type enzyme. The pH activity profiles of wild-type and mutant pepsin is similar. Mutant enzyme has a stronger affinity for the synthetic substrate KPAEFF(NO2)AL. Turnover number for mutant enzyme is significantly lower than that of the wild-type. kcat/Km is 1.9fold lower than wild-type value Sus scrofa
D32E unlike wild-type, the mutant fusion protein is incapable of autocatalytic activation upon acidification of the medium, as determined by Western blot analysis. Mature mutant pepsin is obtained by processing fusion protein samples through an immobilized pepsin column. Tm-value of mutant enzyme is 63°C compared to 71°C for wild-type enzyme. Pronounced decrease in activity below pH 2.5. Mutant enzyme has a stronger affinity for the synthetic substrate KPAEFF(NO2)AL. Turnover number for mutant enzyme is significantly lower than that of the wild-type. kcat/Km is 8.7fold lower than wild-type value Sus scrofa
D32E/D215E unlike wild-type, the mutant fusion protein is incapable of autocatalytic activation upon acidification of the medium, as determined by Western blot analysis. Mature mutant pepsin is obtained by processing fusion protein samples through an immobilized pepsin column. Tm-value of mutant enzyme is 63°C compared to 71°C for wild-type enzyme. The pH activity profiles of wild-type and mutant pepsin is similar. KM-value for KPAEFF(NO2)AL is not significantly different relative to wild-type. Turnover number for mutant enzyme is significantly lower than that of the wild-type. kcat/Km is fold lower than wild-type value. kcat/Km is 13.3fold lower than wild-type value Sus scrofa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.036
-
 KPAEFF(NO2)AL mutant enzyme D215E Sus scrofa
0.04
-
 KPAEFF(NO2)AL mutant enzyme D32E Sus scrofa
0.051
-
 KPAEFF(NO2)AL mutant enzyme D32E/D215E Sus scrofa
0.075
-
 KPAEFF(NO2)AL wild-type enzyme Sus scrofa

Organism

Organism UniProt Comment Textmining
Sus scrofa
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
KPAEFF(NO2)AL + H2O
-
 Sus scrofa ?
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5
-
 KPAEFF(NO2)AL mutant enzyme D32E/D215E Sus scrofa
6
-
 KPAEFF(NO2)AL mutant enzyme D32E Sus scrofa
25
-
 KPAEFF(NO2)AL mutant enzyme D215E Sus scrofa
96
-
 KPAEFF(NO2)AL wild-type enzyme Sus scrofa