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Literature summary extracted from

  • Henot, F.; Pollack, R.M.
    Catalytic activity of the D38A mutant of 3-oxo-DELTA 5-steroid isomerase: recruitment of aspartate-99 as the base (2000), Biochemistry, 39, 3351-3359.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.3.3.1 D38A, D38A/Y14F, D38A/Y55F and D38A/D99N proteins are expressed in Epicurian Coli XL1-Blue cells Comamonas testosteroni

Protein Variants

EC Number Protein Variants Comment Organism
5.3.3.1 D38A the catalytic turnover number is 140fold less than that for the wild-type Comamonas testosteroni
5.3.3.1 D38A/D99N no detectable activity Comamonas testosteroni
5.3.3.1 D38A/Y14F no detectable activity Comamonas testosteroni
5.3.3.1 D38A/Y55F no detectable activity Comamonas testosteroni

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.3.3.1 0.067
-
 5-androstene-3,17-dione 25°C, pH 6.5, solvent D2O, mutant enzyme D38A Comamonas testosteroni
5.3.3.1 0.073
-
 5-androstene-3,17-dione 25°C, pH 8.5, solvent D2O, mutant enzyme D38A Comamonas testosteroni
5.3.3.1 0.085
-
 5-androstene-3,17-dione 25°C, pH 7.5, solvent D2O, mutant enzyme D38A Comamonas testosteroni
5.3.3.1 0.094
-
 5-androstene-3,17-dione 25°C, pH 8.5, solvent H2O, mutant enzyme D38A Comamonas testosteroni
5.3.3.1 0.096
-
 5-androstene-3,17-dione 25°C, pH 7.5, solvent H2O, mutant enzyme D38A Comamonas testosteroni
5.3.3.1 0.099
-
 5-androstene-3,17-dione 25°C, pH 6.5, solvent H2O, mutant enzyme D38A Comamonas testosteroni

Organism

EC Number Organism UniProt Comment Textmining
5.3.3.1 Comamonas testosteroni
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.3.3.1 5-Androstene-3,17-dione residue Asp38 acts as a base to abstract a proton from C-4 of the substrate to form an intermediate dienolate, which is then reprotonated on C-6 Comamonas testosteroni 4-Androstene-3,17-dione
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.3.3.1 230
-
 5-androstene-3,17-dione 25°C, pH 6.5, solvent D2O, mutant enzyme D38A Comamonas testosteroni
5.3.3.1 260
-
 5-androstene-3,17-dione 25°C, pH 8.5, solvent D2O, mutant enzyme D38A Comamonas testosteroni
5.3.3.1 279
-
 5-androstene-3,17-dione 25°C, pH 6.5, solvent H2O, mutant enzyme D38A Comamonas testosteroni
5.3.3.1 340
-
 5-androstene-3,17-dione 25°C, pH 6.5, solvent H2O, mutant enzyme D38A Comamonas testosteroni
5.3.3.1 412
-
 5-androstene-3,17-dione 25°C, pH 7.5 or 8.5, solvent H2O, mutant enzyme D38A Comamonas testosteroni